Journal of Capital Medical University

Correlation among psoriasis vulgaris’ traditional Chinese medicine syndromes, age and course

CHEN Wei-wen;WANG Ping;ZHANG Guang-zhong;JIANG Chun-yan;ZHOU Dong-mei;SUN Li-yun;WANG Ju-sheng;DENG Bing-xu

2011, 32(4): 445-448. doi:10.3969/j.issn.1006-7795.2011.04.001

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Objective To explore the correlation among psoriasis patients’ age of onset, age, recurrence frequencies , disease course and traditional Chinese medicine(TCM) syndromes. Methods Based on the clinical epidemiologic mass survey, the study was carried out by means of a status survey, multi-center and large-sample research. The data base was set up by EPIINFO 6.0. The SPSS was used to do the statistical analyses in 2 675 cases of psoriasis vulgaris to study the correlations among patients’ age of onset, age, recurrence frequencies, disease course and TCM syndromes. Results The TCM syndromes in psoriasis vulgaris mainly include the blood heat syndrome 1 489(55.7%), blood dryness syndrome 593(22.2%), and blood stasis syndrome 323(12.1%). The sequences from young to old in age of onset and age are blood heat syndrome, blood dryness syndrome, and blood stasis syndrome(P<0.01). Blood heat’s recurrence frequencies is the least among the 3 syndromes(P<0.01). The sequences from short to long of disease courses are blood heat syndrome, blood dryness syndrome, and blood stasis syndrome(P<0.01). Conclusion The blood heat syndrome, blood dryness syndrome, and blood stasis syndrome are the main syndromes of psoriasis vulgaris. There are not only onset sequences but also different degrees of treatment difficulty among the 3 TCM syndromes.

Optimization of Alu-LTR PCR approach to detect integrated HIV-1 DNA in variety of cell subsets at different stages after HAART

LI Juan;JIAO Yan-mei;GAO Yan-qing;KOU Bu-xin;ZHANG Tong;WU Hao

2011, 32(4): 449-452. doi:10.3969/j.issn.1006-7795.2011.04.002

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Objective To determine if there is integrated human immunodeficiency virus-1(HIV-1) DNA in variety of cell subsets at different stages after highly active antiretroviral therapy(HAART) through detecting HIV-1 DNA in CD4⁺T cell, CD8⁺T cell and B cell by Alu-long terminal repeat(LTR) polymerase chain reaction(PCR). Methods The genomic DNA from purified CD4⁺T, CD8⁺T, B cells of 20 HIV-1 patients after HAART 0, 4, 12 W and 10 healthy control subjects was obtained by Alu-LTR PCR. Results After Alu-LTR PCR, integrated HIV-1 was detected in each of the 20 cases of HIV/AIDS of CD4⁺T cells and CD8⁺T cells after HAART. The product of CD8⁺T cells were lower than that of the CD4⁺T cells. The product of HAART 4, 0, 12 W had no significant difference. Conclusion There exists integrated HIV-1 DNA in CD4⁺T cells and CD8⁺T cells, and HIV viral reservoir mainly exists in CD4⁺T cells. Integrated HIV-1 DNA do not exist in B cells. Integrated HIV-1 DNA still exist, and HAART do not eradicate HIV viral reservoir.

Influence of early syphilis on CD4 cell count in human immunodeficiency virus infection

HUANG Shu-yan;WU Hao;ZHANG Ming;SUN Xin;HUANG Xiao-jie;JIAO Yan-mei;LI Juan;GAO Yan-qing

2011, 32(4): 453-455. doi:10.3969/j.issn.1006-7795.2011.04.003

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Objective To assess the effect of early syphilis on CD4 cell count among men who have sex with men (MSM) with human immunodeficiency virus(HIV) infection. Methods Cases of early syphilis in HIV infected patients among MSM were included. Benzathine penicillin was given after the diagnosis of syphilis. CD4 cell counts were measured by flow cytometry before, during, and after three and six months of treatment, and the results were compared. Results Fifty patients were included, and forty-five patients had successful treatment of syphilis. CD4 cell count decreased significantly during infection in patients with early syphilis(P=0.04). There was no significant difference in the levels of CD4 cell during infection and after the 3-month treatment(P>0.05), and CD4 cell count increased significantly during infection and after the 6-month treatment(P=0.02). Conclusion CD4 cell count decreased in the early infection with Treponema pallidum in HIV infected patients, and the successful treatment of syphilis can reverse the trend.

Skin infection of newborn infants in maternal-neonatal unit

MA Xiu-hua;SONG Feng-li;MA Li-li;SHI Xin;LI Yan;SHI Wei;WANG Chun-xia;LIU Ying;MA Lin

2011, 32(4): 456-459. doi:10.3969/j.issn.1006-7795.2011.04.004

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Objective To investigate the causes and the characteristics of skin infection of newborn infants in maternal-neonatal unit and its control measures. Methods Totally 9 131 newborns in the nursery of the department of obstetrics from January 2009~December 2010 were investigated. Results A total of 206 newborn infants klebsiella pneumoniae and staphylococcus epidermidis. staphylococcus aureus isolates were sensitive to vancomycin, paediatric compound sulfamethoxazole, piperacillin-tazobactam and linezolid, while were resistant to penicillin, ampicillin and erythromycin. Conclusion The characteristics of the newborn skin infection were associated with the internal causes of neonates and environmental factors. The occurance of infection is related to the mode of delivery, labor process and complications in the prenatal period. Attention should be paid to pregnancy health care, reduction of the risk factors for skin infection in newborn infants in maternal-neonatal unit to effectively control neonatal skin infections.

A survey of mixed infection with pathogens in sexually transmitted disease clinic

KE Wu-jian;CHE Ya-min;TIAN Jing-qun;FENG Bin;LI Chun-li;ZHANG Tian-lu

2011, 32(4): 460-463. doi:10.3969/j.issn.1006-7795.2011.04.005

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Objective To investigate the epidemiological characteristics of mixed infection of the pathogens of sexually transmitted disease(STD) in patients attending STD clinic. Methods Totally 1,124 cases with STD were tested for Chlamydia trachomatis(CT), Ureaplasma urealyticum(UU), Treponema pallidum(TP), human papillomavirus (HPV), Candida albicans(CA), Neisseria gonorrhoeae(NG) and human immunodeficiency virus (HIV), respectively. Results The mixed infection rate was 36.0%(405/1 124). Sixteen kinds of mixed infection were found. Mixed infections were predominant in the 21 to 30 age group, high school or less educated people, workers and self-employed persons. Compared with non-mixed-infected individuals, the difference of gender, average age and marital status in subjects with mixed infection were not statistically significant(P>0.05). The educational status in subjects with mixed infection was lower, while the average number of sexual partners was larger than that in single pathogen-infected people(P=0.001). Conclusion The mixed infection rate in STD clinic is quite high and the mixed infection pattern was diverse. The mixed infection was correlated with low education and multiple sex partners.

Effects of intense pulsed light on matrix metalloproteinases-1, -8 and tissue inhibitor of metalloproteinase-1 from human skin

YUAN Ning;ZHAO Jun-Ying

2011, 32(4): 464-468. doi:10.3969/j.issn.1006-7795.2011.04.006

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Objective To analyze quantitatively the alteration of matrix metalloproteinases-1, -8(MMP-1, MMP-8), tissue inhibitor of metalloproteinase-1(TIMP-1) after radiation with intense pulsed light(IPL) technique in clinic. And to partly explain the treatment mechanism of IPL on sun-damaged human skin and provide a theoretical basis of mechanism, course of therapy and interval time for treatment of IPL on photo aging skin. Methods Volunteers older than 20-years of age whose skin type is Fitzpatrick Ⅲ~Ⅳ were selected. One site of each volunteer was radiated respectively with IPL system at different parameters. Skin specimens were taken at both non-treated and treated locations on days 14 and 21 after IPL irradiation of about the size of 5 mm×5 mm×5 mm. All skin specimens were fixed in formaldehyde fixative buffer. Immunohistochemical staining was performed on each specimen, including observations on changes of MMP-1, MMP-8, TIMP-1 proteins expression after IPL irradiation in order to know their action in dermis remodeling. Results The expressions of MMP-1, MMP-8 and TIMP-1 were detected after treatments. To weeks 2 after treatment. The expressions of MMP-1, MMP-8 became weaker at weeks 3. Expression of TIMP-1 was not significantly different between weeks 2 and 3. There were no expressions in normal skin. Conclusion IPL irradiation could promote the expression of MMP-1, MMP-8 and TIMP-1 in human skin directly.

Reverse line blot hybridization and DNA sequencing studies of the 16S~23S rDNA gene internal transcribed spacer regions of five emerging pathogenic Nocardia species

WANG Xiao-yan;KONG Fan-rong;LIAN Shi

2011, 32(4): 469-475. doi:10.3969/j.issn.1006-7795.2011.04.007

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Objective To examine DNA sequence polymorphisms in the 16S~23S rDNA gene internal transcribed spacer(ITS) regions of five emerging species: Nocardia beijingensis, N. blacklockiae, N. thailandica, N. elegans and N. vinacea and to screen specialized Nocardia ITS probes for epidemiological study. Methods A set of six isolates belonging to the species of interest was studied. A PCR-based reverse line blot(RLB) hybridization assay incorporating species, or intraspecies ITS rDNA gene operon-specific probes was then developed for species identification. Results Substantial intraspecies sequence variation among different ITS operons was identified. Four sequence types of N. thailandica, eight types of N. beijingensis, and five sequence types of N. blacklockiae, N. elegans and N. vinacea were found. A number of unique RLB pattern types were observed for the cloned products of 2 N. beijingensis species including 4 RLB types, N. blacklockiae 3, N. elegans 3, N thailandica 4 and N. vinacea 2. Conclusion The results represent the first evidence of ITS sequence heterogeneity in emerging species of Nocardia. The PCR/RLB assay demonstrated high specificity and showed promise in both the identification and genotyping of Nocardia species. All N. beijingensis-ITS, N. blacklockiae-ITS and N. elegans-ITS probes were highly specific.

Present status of research on pruritic papular eruption in patients with HIV infection

HAN Jie;ZHU Wei;LUN Wen-hui;LIAN Shi

2011, 32(4): 476-478. doi:10.3969/j.issn.1006-7795.2011.04.008

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Pruritic papular eruption of HIV(HIV-PPE) has become the common cutaneous manifestation in HIV-infected patients. HIV-PPE has a higher prevalence in tropic regions. PPE is characterized by chronic pruritus and symmetric papular eruptions. However, there is no clear consensus on the etiology of PPE and the pathologic findings. PPE is correlated with decrease in CD4 cell counts. The effective treatments for PPE are unavailable at present.

Advances in studies on the plant extracts’ whitening efficacy

CUI Wen-ying;YANG Gao-yun;LI Yan

2011, 32(4): 479-483. doi:10.3969/j.issn.1006-7795.2011.04.009

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This review article presents the mechanism of plant extracts’ whitening effect from reduction of melanin formation, keratolytic, natural sunscreens, inhibiting melanin formation at genetic level, reduction of the melanin transference. This article also summarizes the present plant extracts’ whitening agents, such as kojic acid, plant polyphenols, phloretin, vitamin C and derivatives, endothelin antagonists, plant flavonoids, as well as the developmental trend of plant extracts from nanotechnology, compositional formulation and herbal cosmetics in the future.

Effects of P165 and rosiglitazone on the expression of IRS-1 and p-CREB in SH-SY5Y cell

ZHANG Jing-yan;WANG Rong;ZHAO Jing-shu;JI Zhi-juan;ZHAO Zhi-wei;SHENG Shu-li

2011, 32(4): 484-488. doi:10.3969/j.issn.1006-7795.2011.04.010

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Objective To investigate the effects of P165(the analog of APP5 peptide) and rosiglitazone on the expression of insulin signaling pathway related factors IRS-1 and p-CREB in SH-SY5Y cells after streptozotocin(STZ)-induced injury. Methods Human neuroblastoma cell line SH-SY5Y cells were divided into four groups: control group, STZ injury group(STZ 0.8 mmol/L), rosiglitazone protection group(STZ 0.8 mmol/L+rosiglitazone 20 μmol/L) and P165 protection group(STZ 0.8 mmol/L+P165 30 μmol/L), and were observed for morphology of cells and the immunofluorescence staining of IRS-1 and p-CREB, and we determined mean optical density using Image-pro plus software. Results Compared with the control group, cells of STZ injury group grew and divided slower, cell processes were shorter and the cell body shrunk, the intensity of immunofluorescence staining of IRS-1 and p-CREB were reduced. Compared with STZ injury group, the cells in two protection groups had neurite extension and ameliorated in cell body shrinkage and morphology, and showed stronger immunofluorescence staining. The mean optical density of IRS-1 and p-CREB in STZ injury group decreased than that of the control group(P<0.008), and the density in two protection groups increased compared with STZ injury group (P<0.008). Conclusion P165 and rosiglitazone could ameliorate the STZ-induced injury to SH-SY5Y by improving the expression of IRS-1 and p-CREB, and thereby protect the neurons.

The effects of remote ischemic post-conditioning on expression of amyloid precursor protein in rats following cerebral ischemia and reperfusion

YAN Feng;LIU Xiang-rong;ZHANG Ying;ZHANG Chen-cheng;JI Xun-ming;LUO Yu-min

2011, 32(4): 489-493. doi:10.3969/j.issn.1006-7795.2011.04.011

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Objective To investigate the effects of remote ischemic post conditioning on the expression of amyloid precursor protein(APP) in rats following cerebral ischemia and reperfusion. Methods The focal ischemia and reperfusion rat model was established with suture method. The remote ischemic post-conditioning was performed with bilateral occluding of bilateral femoral arteries for 10 min and reperfusion for 10 min, repeated for 3 times. The rats were divided into 7 groups: sham group(n=4, named S), control group(n=8, rats were only subjected to focal cerebral ischemia and reperfusion, 4 rats were sacrificed at 4 hours after ischemia and another 4 at 24 hours, named C4, C24), and the experiment groups(n=8, the rats underwent focal cerebral ischemia/reperfusion and remote ischemia post conditioning immediately after cerebral ischemia: 4 rats were sacrificed at 4 hours after ischemia and the other 4 at 24 hours, named I4, I24; n=8, rats underwent focal cerebral ischemia/reperfusion and remote ischemia post conditioning immediately after reperfusion; 4 of the rats were sacrificed at 4 hours after ischemia and the other 4 at 24 hours, named R4, R24). Beta-APP was determined quantitatively by Western blotting. The immunofluorescence was used for the localization of β-APP. Results There were no significant differences between C4, C24 and S groups. But the level of β-APP in the C24 group was increased compared to the S group. Compared to the C24 group, the level of β-APP was significantly decreased in the I24 group(P<0.05). Conclusion Remote ischemic post-conditioning might decrease the level of β-APP induced by cerebral ischemia, resulting in attenuation of cerebral ischemic damage.

Effects of flavonoids from scutellaria stem and leaf on memory impairment and the expression of choline acetyltransferase and nitric oxide synthase in chronic cerebral ischemia rats

WANG Yu-mei;CAO Kai;LIU Yong-ping;QIN Bo-wen;SHANG Ya-zhen

2011, 32(4): 494-500. doi:10.3969/j.issn.1006-7795.2011.04.012

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Objective To study the effects of flavonoids from scutellaria stem and leaf(SSF) on memory impairment and the expression of choline acetyltransferase(ChAT) and nitric oxide synthase(NOS) in chronic cerebral ischemia rats. Methods The model of chronic cerebral ischemia was established by bilateral carotid artery occlusion for 2 months with female SD rats. The functions of learning and memory were detected by Morris water maze, the expression of ChAT, neuronal nitric oxide synthase(nNOS), inducible nitric oxide synthase(iNOS) and endothelial nitric oxide synthase(eNOS) were examined by immunohistochemical method. Results Compared with the sham group, memory ability was significantly decreased, the expressions of ChAT and eNOS significantly decreased, as well as the expressions of nNOS and iNOS were dramatically increased in cerebral ischemia group rats. However, the treatment with 17.5, 35 and 70 mg·kg^-1 SSF for 38 d of the ischemic rats showed a significant improvement in impairment of memory ability and the abnormal changes of ChAT, eNOS nNOS and iNOS in the brain of ischemic rats. Conclusion SSF can improve the memory impairment induced by chronic cerebral ischemia in rats, which may be related to regulation of the abnormal changes of ChAT and NOS in the ischemic rats’ brain.

Influence of peptidoglycan on BV2 cells after endocytosis of amyloid protein β oligomers

JIANG Xin;XIANG Rong-cai;BAI Li-juan;ZHANG He-min;CHEN Xiao-hong;MA En-long

2011, 32(4): 501-508. doi:10.3969/j.issn.1006-7795.2011.04.013

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Objective To investigate the influences and mechanisms of the pro-inflammatory medium of peptidoglycan(PGN) on BV2 cells after endocytosis of amyloid protein β(Aβ) oligomers. Methods BV2 cells were cultivated by cell passage method, instead of neurons and microglia separately. Aβ_1-42 oligomers were prepared according to Klein WL (2002). The endocytosis amount of Aβ in BV2 cells was determined by immunofluorescence staining. The expression of phosphorylated p38 mitogen-activated protein kinase(p-p38MAPK), p38MAPK protein of BV2 cells in each group was determined by Western blotting method, and mouse homologue formyl peptide receptor 2(mFPR2)mRNA by PCR method. Results The endocytosis of Aβ42 oligomer increased after BV2 cells was activated by PGN, which could be inhibited by mFPR2. PGN could lead to the increase of mFPR2 mRNA expression in BV2 cells in relation with concentration and time, which could be inhibited by SB202190-p38MAPK. When the expression of SB202190 increased, the inhibition increased. Compared with 0 μmol/L, concentration of each group was inhibited more significantly, for 1 μmol/L group(P<0.05), for 10, 20, 30 μmol/L groups(P <0.01). After PGN activated BV2 cells, the activation of p38MAPK phosphorylation at each time point was significantly different from the control group(P<0.01). Conclusion The endocytosis of Aβ42 oligomer increased after BV2 cells was activated by PGN, which can be inhibited by mFPR2, so it is speculated that endocytosis of Aβ in BV2 cells may have relation with the expression of mFPR2. PGN could lead to the increase of mFPR2 mRNA expression in BV2 cells, which may participate in the endocytosis of Aβ after BV2 cells are activated. PGN can lead to the increase of p38MAPK expression in BV2, and its inhibitor can decrease the expression of mFPR2 mRNA, which may be the mechanism of endocytosis of Aβ after BV2 cells are activated.

Influences of peptidoglycan on PC12 cells after endocytosis of amyloid protein β oligomers in activated BV2 cells

JIANG Xin;XIE Ming;BAI Li-juan;CHEN Xiao-hong;MA En-long

2011, 32(4): 509-513. doi:10.3969/j.issn.1006-7795.2011.04.014

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Objective To investigate how the pro-inflammatory medium of peptidoglycan(PGN) acts on PC12 after endocytosis of amyloid protein β(Aβ)oligomers in activated BV2 cells. Methods PC12 cells and BV2 cells, instead of neurons and microglia separately, were co-cultivated in transfer screen by cell passage method. Aβ_1-42 oligomers were prepared according to Klein WL(2002). The inhibition of PC12 cells was determined by MTT, and the expression of its tau[ps396] protein was determined by Western blot, and the apoptosis of PC12 cells was determined by flow cytometry. Results Aβ oligomers, BV2 cells acted upon by Aβ oligomers, and BV2 cells affected by PGN all could restrain the proliferation of PC12 cells, increase the expression of its tau[ps396], and increase its apoptosis, while the effect of PGN after endocytosis of Aβ oligomers in activated BV2 cells were more remarkable. Conclusion Aβ oligomers can increase the inhibition of PC12 cells, abnormal phosphorylation of tau protein, and the apoptosis, and after endocytosis in activated BV2 cells by PGN, the effect of Aβ oligomers is more remarkable.

Recent progress in caloric restriction and the mechanism of its neuroprotective effects

ZHAO Jing-shu;WANG Rong

2011, 32(4): 514-518. doi:10.3969/j.issn.1006-7795.2011.04.015

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Caloric restriction(CR) is a kind of dietary mode that decreases calories intake by 30%~50% under the premise of nutritional adequacy. It has aroused increasing attention that CR can extend lifespan of organism, delay or prevent the occurrence and development of several ageing-related diseases. There are several mechanisms proposed to explain the neuroprotective effects of caloric restriction: antioxidant effects, increased metabolic efficiency, increased sirtuin and neurotrophic factor activity, and anti-inflammatory effects.

Late endosome sorting contributed to the transduction activation of type 2 adeno-associated virus in IB3 transformed human bronchial epithelial cells

YAN Shao-fei;WEN Zhao-yang;GAO Er-jing;NIU Jing;ZHENG Shao-peng;DING Wei

2011, 32(4): 519-524. doi:10.3969/j.issn.1006-7795.2011.04.016

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Objective To test whether type 2 adeno-associated virus(AAV2) is able to undergo vesicular sorting and traffic into the late endosome, and then evaluate the endosome-contained AAV2 possess different transduction activities, thus to imply the role of endosomal trafficking in the activation of AAV2 transduction in transformed human bronchial epithelial cells. Methods Using the RAB7 GFP fusion protein as the specific late endosome marker, its colocalization with AF568 fluorochrome labeled AAV2 was analyzed by confocal microscopy. The infected AAV2 was retrieved by gradient centrifugation and magnetic beads-based immunoisolation together with the endosome, and then subjected to the reinfection of IB3 cells. The efficiency of the AAV2 transgene expression was compared under the conditions without and with the modulation of RAB7 functions. Results It was found that a significant portion of the endocytosed AAV2 was able to traffic into the late endosome in IB3 cells. The AAV2 recovered from the late endosome exerted better transduction activities than the native viruses. Conclusion The results implied that the vesicular sorting of AAV2 into the late endosome might be an important step for the activation of the AAV mediated transgene expression.

Effects of overexpression of nurr1 on apoptosis of SK-N-SH cells induced by neurotoxin 6-OHDA

LIU Yang;ZHAO Yong-mei;ZHANG Hai-yan;LI Wei-hong;YUE Yun

2011, 32(4): 525-533. doi:10.3969/j.issn.1006-7795.2011.04.017

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Objective To investigate the possible correlation between the expression of Nurr1 gene, a midbrain transcription factor, and selective apoptotic cell death induced by neurotoxin 6-OHDA. Methods Firstly, the expression of Nurr1 was detected by immunohistochemistry and RT-PCR in transfected SK-N-SH/Nurr1 cells. Secondly, following treatment with 50~100 μmol/L 6-OHDA on both SK-N-SH and SK-N-SH/Nurr1 cells for 6 and 12 h, the cell loss was detected to discriminate between apoptosis and necrosis by using flow cytometry(FCM) and Annexin V/PI double staining. Thirdly, the ultrastructural changes of both cells were observed by transmission electronic microscopy(TEM) after treatment with 75 μmol/L 6-OHDA for 12 h. Finally, the level of apoptosis effector Caspase-3 of both cells were detected after exposure to 75 μmol/L 6-OHDA for 6 and 12 h by Western blotting analysis. Results Firstly, immunohistochemical staining and RT-PCR analysis showed that the over expression of Nurr1 in transfeced SK-N-SH cells. Secondly, Annexin V/PI double staining with FCM showed treatment with 75 μmol/L 6-OHDA for 12 h induced a significant apoptotic phase with Annexin V+/PI-only in SK-N-SH/Nurr1 cells(P=0.000), while both cells showed increased necrotic phase with Annexin V+/PI+ after exposure to 100 μmol/L 6-OHDA for 6 and 12 h. Thirdly, the result of TEM showed that treatment with 75 μmol/L 6-OHDA for 12 h induced typical apoptosis in SK-N-SH/Nurr1 cells. In contrast, no morphological characteristics of apoptosis in SK-N-SH cells were observed. Finally, Western blot analysis showed that treatment with 75 μmol/L 6-OHDA for 6 and 12 h decreased the level of pro-Caspase-3 in SK-N-SH/Nurrl cells but not in SK-N-SH cells(P=0.000). Conclusion Nurr1-overexpression stimulated apoptotic pathway induced by 6-OHDA,specifically in a dose-dependence manner.

High performance liquid chromatographic determination of adenosine phosphates in the mouse brain

WU Yan-chuan;LI Yao-hua;XIE Sheng-nan;LI Xin;YU Shun;He Shi-da

2011, 32(4): 534-537. doi:10.3969/j.issn.1006-7795.2011.04.018

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Objective To establish a method for the quantitative determination of adenosine phosphates in mouse brain by high performance liquid chromatography-variable wavelength detector(HPLC-VWD). Methods The protein in mouse brain tissue sample was precipitated with perchloric acid. Separation was performed on a reverse phase Supelco Discovery C18 column(150 mm×4.6 mm, 5 μm), equipped with a pre-column of the same material by Agilent 1200 HPLC-VWD. The wavelength was 254 nm. The mobile phase contained methanol-30 mmol/L potassium dihydrogen phosphate buffer, pH 5.85(5∶95, v/v) . The flow rate was set at 1.0 mL/min and the injection volume was 20 μL. The column temperature was 30 ℃. Results The linear relationship between the range(10~100) mg/L was good. The linear equations were ATP: A=28.924C+14.144, γ=0.999 90; ADP: A=31.504C-12.608, γ=0.999 92; AMP: A=46.539C-5.255, γ=0.999 95. The average recovery rate was 95.3%~103.2%. The RSDs of intra-and interday were 1.14%~1.75% and 2.45%~3.97%, respectively. Conclusion The experimental data proved that the HPLC-VWD method could detect the concentration of ATP, ADP and AMP in the mouse brain on repeated hypoxic exposures accurately, rapidly and simply.

Purification of asiatic acid from centella asiatica and its determination by RP-HPLC

XIA Bin-bin;LI Yi-sha;XU Wei-zhe;WANG Qiao;SONG Xue-ying

2011, 32(4): 538-540. doi:10.3969/j.issn.1006-7795.2011.04.019

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Objective To establish a rapid separation and purification method of asiatic acid from centella asiatical(L.) Urban and to develop a reversed-phase high-performance liquid chromatography(HPLC) method with UV-vis detection for the quantitative determination of asiatic acid. Methods Crude extract separated and purified by silica gel column chromatography with gradient elution of petroleum ether-acetone system was first extracted with ethanol from centella asiatical(L.) urban and then done by ethyl acetate. TLC was used as the monitor during the process. HPLC separation was achieved on a Symmetry C₁₈ column(4.6 mm×250 mm, 5 μm) with a solvent system of acetonitrile -10 mmol·L^-1 ammonium acetate(38∶62). The wavelength of detector was set at 210 nm, the flow rate was 1 mL·min^-1, the column temperature was set at 25 ℃, and the injection volume was 20 μL. Results A calibration curve ranging from 10 to 200 mg·L^-1 was shown to be linear(R²=0.999 5), and the lower limit of quantification(LLOQ) was 10 mg·L^-1. The inter- and intra-day RSD% which were determined by three different concentration(25, 100, 200 mg·L^-1) were lower than 5.4%. The content of asiatic acid in centella asiatical(L.) Urban was 0.99 g·kg^-1. Conclusion The method was successfully applied to the separation, purification and the quantitative determination of AA.

Weight-losing effect of a novel fatty acid synthase inhibitor from extract of Pangdahai on rats with diet-induced obesity

GAO Li-fang;CAO Li-ge;TIAN mi;GAO Wen;CHEN Zhen-liang

2011, 32(4): 541-544. doi:10.3969/j.issn.1006-7795.2011.04.020

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Objective To investigate the effect of sterculia scaphigera(Pangdahai) extract on fatty acid synthase and weight reduction in diet-induced obesity rats. Methods SPF-grade adult male Wistar rats were fed with high-fat diet, at the same time an extract from Pangdahai 10, 30 and 100 mg/kg were administered by oral gavage once daily respectively, which lasted for 45 d. At the end of administration, the body weight(BW), adipose and food consumption were recorded, the content of serum biochemistry and the activity of hepatic fatty acid synthesis(FAS) were measured. Results Compared with the model-control group, the BW, adipose, food intake and the hepatic FAS activity were significantly decreased in the Pangdahai extract 100 mg/kg intervention group(P<0.05). Conclusion Pangdahai extracts at the dose of 100 mg/kg could inhibit FAS activity and appetite of the rats.

Specificity and sensitivity and expression of glypican-3 in hepatocellular carcinoma

ZHANG Shi-jie;LANG Zhen-wei;WANG Xin-xin;Lü Fu-dong

2011, 32(4): 545-548. doi:10.3969/j.issn.1006-7795.2011.04.021

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Objective To investigate specificity, sensitivity and expression of glypican-3(GPC3) in well differentiated, moderately or poorly differentiated hepatocellular carcinoma(HCC) groups Methods Seventy cases with HCC were chosen from our hospital from Dec. 2009 to Sep. 2010. Expression of GPC3 in HCC as well as in non-HCC surrounding tissue was observed by immunohistochemical-staining and detection rate of GPC3 was calculated. Seventy cases were divided into two groups, well differentiated and moderately/poorly differentiated groups. Correlation between GPC3 and Ki67 were analyzed in two groups. Results 1 Staining for GPC3 was positive only in HCC tissues, and was negative in non-HCC surrounding tissue of HCC. Detection rate of GPC3 was 85% in two groups, positive rate 73% in the well differentiated group, 90% in the moderately/poorly differentiated group. 2 Staining intensity was middle/strong for GPC3 in two cases, accounting for 10% in the well differentiated group, 38 cases(74.8%) in the mediately/ poorly differentiated group. Expression of GPC3 was significantly different in well differentiated group and moderately/poorly differentiated group(P<0.01). 3 In none of the cases the expression level of the Ki67 was moderate and strong(++, +++). In 37 cases(72%) of the moderately/poorly differentiated group, the expression was moderately strong. Expression of Ki67 was significantly different in well, moderately/poorly differentiated group(P<0.01). Staining intensity of GPC3 was correlated with expression of Ki67(P<0.01). Conclusion Expression of GPC3 is of the high degree of specificity and sensitivity in HCC. GPC3 may have important value in pathological diagnosis of HCC.

Expression and significance of β-catenin, cyclinD1, c-myc in breast carcinoma

HE Yan-jiao;QIAO Chao;LIU Zhao-xia;YU Jin;LI Guang

2011, 32(4): 549-552. doi:10.3969/j.issn.1006-7795.2011.04.022

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Objective To study the expression and significance of β-catenin, cyclinD1, c-myc in breast carcinoma. Methods Breast tissue specimens from 119 patients with breast carcinomas, 72 patients with breast cystic hyperplasia and 40 normal individuals were studied. Immunohistochemistry was used to detect the expressions of β-catenin, cyclinD1, c-myc, then their relationship with the clinicopathological features of patients with breast carcinoma was analyzed. Results In normal breast tissues, from the 40 control epithelial cells showed equally strong membranous expression of β-catenin protein at the cell-cell boundaries, but the expression of cyclinD1, c-myc was negative. The abnormal rate of β-catenin in breast carcinoma(78.15%) was much higher than that in patients with breast cystic hyperplasia tissues(44.44％), significant correlation was found(P<0.01). The overexpression of cyclinD1, c-myc in breast carcinomas(57.98%, 56.30%) was much higher than that of breast cystic hyperplasia tissues(33.33%, 27.78%), significant correlation was found(P<0.01). The abnormal expression of β-catenin and the overexpression of cyclinD1 was associated with lymph node metastasis(P=0.031, P=0.003). The overexpression of c-myc was associated with size and lymph node metastasis(P=0.041). There was a highly significant association between abnormal expression of β-catenin and overexpression of cyclinD1(P=0.006), but not between abnormal expression of β-catenin and overexpression of c-myc(P=0.775). Conclusion The abnormal expression of β-catenin may play an important role in the carcinogenesis and progression of human breast carcinoma by up-regulating the expression of cyclinD1. β-catenin abnormal expression and the cyclinD1, c-myc overexpression could be considered as useful makers for determining metastasis and prognosis of human breast carcinoma

Effect of Nicorandil preconditioning of rats on storage of their heart exvivo

ZHANG Guo-bin;LIU Ya-nan;ZHANG Yu

2011, 32(4): 553-557. doi:10.3969/j.issn.1006-7795.2011.04.023

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Objective To explore the mechanism of protecting effect of Nicorandil on isolated heart of rats in cold preservation solution. Methods A total of 24 SD male rats were randomly divided into a control group and nicorandil group, in which the rats were injected intraperitioneally with distilled water or nicorandil 0.1 mg·kg^-1·d^-1 for 9 days respectively. After the last injection, every isolated heart was stored in 4 ℃ St-Thomas solution. The myocardial tissue was taken from the anterior wall of the left ventricle at 2, 4, 6, 8, 10, 12, 16, 20 and 24 hours respectively. The contents of MDA and the changes of ultrastructure of myocardial tissue were observed. The protein expressions of HSP70 were examined by using immunohistochemical staining and Western blotting. Results The ultrastructure of myocardial tissue changed more significantly in the nicorandil group. MDA concentrations were reduced significantly in the nicorandil group compared with those of the control group(P<0.05). With the time prolonged, the expression of HSP70 increased gradually and dropped later as parabola-related curves in nicorandil group compared with the basement expression of HSP70 in control group respectively. The peak expression of HSP70 was seen at 6 hours and significantly decreased at 24 hours in nicorandil group. The correlation between MDA and HSP70 are significant(P<0.05). Conclusion Nicorandil preconditioning may have a protective effect in the isolated rat heart by the induction of the early endogenous HSP70 expression.

Changes in platelet volume indices in patients with Graves’ disease before and after treatment

YANG Ning;XU Yuan;ZHANG Dong-lei

2011, 32(4): 558-561. doi:10.3969/j.issn.1006-7795.2011.04.024

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Objective To study the changes in platelet volume indices in patients with Graves’ disease(GD) before and after therapy. Methods From July 2009 to Dec. 2009, 275 patients with GD were enrolled in this study, 199 patients were euthyroid(Euthyroid group), 99 patients were still hyperthyroid(Hyperthyroid group). Platelet count(PLT), mean platelet volume(MPV), platelet distribution width(PDW), platelet-large cell ratio(P-LCR) and changes of mean platelet volume(△MPV) and platelet-large cell ratio(△P-LCR) were measured. Results Before therapy, compared to patients with euthyroid state, MPV and P-LCR were significantly lower in patients with hyperthyroid state. After therapy, MPV, PDW and P-LCR were significantly higher in the hyperthyroid group. Before and after therapy in two groups, PDW, △MPV and △P-LCR were significantly different(P<0.01). P-LCR was higher than 24.85% before therapy and lower than 30.55% after therapy, which suggest that GD patients may have been in euthyroid state after therapy. Conclusion GD patients achieved euthyroid state after treatment, there were falls in MPV, PDW and P-LCR than before. There were significant differences in PDW, △MPV and △P-LCR in the two groups before and after therapy. P-LCR suggested that the therapy may be effective and it maybe features of GD that have not previously been described.

Relationship between retinopathy in type 2 diabetics and glycated hemoglobin

MA Jin-li;ZHU Cheng-yi

2011, 32(4): 562-564. doi:10.3969/j.issn.1006-7795.2011.04.025

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Objective To explore the relationship between retinopathy and HbA1c in type 2 diabetics. Methods Totally 300 cases with type 2 diabetes mellitus were divided into 3 groups: non-diabetic retinopathy group(NDR), nonproliferative diabetic retinopathy group(NPDR) and proliferative diabetic retinopathy group(PDR) according to the degree of retinopathy. Meanwhile its correlated risk factors including HbAlc, age, gender, duration of diabetes, and blood pressure were statistically analyzed. Results There was statistically significant difference in the level of HbA1c among the three groups(P<0.05). Conclusion The level of HbA1c is one of important indicators of diabetic retinopathy.

Target controlled infusion of remifentanil in patients undergoing cystoscope

WEI Wei;TIAN Ming

2011, 32(4): 565-568. doi:10.3969/j.issn.1006-7795.2011.04.026

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Objective To estimate the feasibility and safety of monitored anethesia using target controlled infusion of remifentanil in patients undergoing cystoscopy. Methods Thirty patients〔American Society of Anesthesiologists(ASA)Ⅰ~Ⅱ grade〕, scheduled for cystoscope, were randomly allocated into two groups. All the patients were treated with target controlled infusion of remifentanil. The target concentrations were 3 μg/L in group L(n=15) and 4 μg/L in group H(n=15) respectively. Operations began as soon as the effective concentration reached preconcerted value. Bispectral index(BIS) was applied in both groups. When patients’ actions appeared that disturbed the operation, higher level of remifentanil could be administered. Oxygen was given by mask routinely and to make breath manually when pulse oxygen saturation(SpO₂)<90%. Blood pressure, heart rate, SpO₂ and BIS value were monitored. Other parameters such as visual analgesia score(VAS), complications, dosage and frequency of remifentanil injection were also analyzed. Results BIS value of the two groups were 89±10 and 86±13 respectively,and the average VAS were 4.8 and 3.3 respectively, which showed good analgesic effects and were not significantly different. Manually assisted respiration was applied in 2/15 cases in group L and in 8/15 cases in group H(P=0.02), respectively; the incidence of body action were 5/15 in group L and 1/15 in group H(P=0.08) respectively; total dosage of propofol given was(125.5±77.7)μg in group L and (242.5±72.6)μg in group H; all of the 5 parameters mentioned above were significantly different between the two groups. Conclusion The schemes of MAC in two groups are in enough depth of anesthesia in patients undergoing cystoscope. The concentration of 2~4 μg/L for remifentanil is considered as safe and effective, with less adverse effects.

Analysis of 9 cases with acute fatty liver in pregnancy

SONG Wei;WANG Xin

2011, 32(4): 569-571. doi:10.3969/j.issn.1006-7795.2011.04.027

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Objective To detect the clinical characteristics of acute fatty liver in pregnancy. Methods Retrospective analysis was performed on 9 cases delivered in Beijing Obstetrics and Gynecology Hospital during the recent 6 years. Results Acute fatty liver in pregnancy occured in the third trimester, the first symptom of the 9 cases was the digestive tract symptom, such as nausea, vomiting, malaise, etc. The damage of liver, kidney and coagulatory system would appear within short time. Liver function abnormality included hypoalbuminemia and elevated hepatic aminotransferase levels(ALT, AST), total bilirubin and direct bilirubin. Kidney function abnormality included elevated BUN and creatinine. Clotting dysfunction included prolonged prothrombin time, low platelet and disseminated intravascular coagulation(DIC). The mortality of pregnant women with this syndrome was 11.1%, and the mortality of neonates was 16.7%. Conclusion The outcome of pregnant women and neonates was closely related to the course of the illness and therapy. Early diagnosis, termination of pregnancy in time, and prevention of DIC would improve the outcome of pregnant women and neonates.

Clinic visiting rate of medical students and its influencing factors

HOU Jie;ZHU Xian

2011, 32(4): 572-575. doi:10.3969/j.issn.1006-7795.2011.04.028

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Objective To analyze the health condition and the influencing factors of medical students and to provide the basis for the medical schools in development of corresponding measurements focusing on health education of students. Methods Based on clinic visiting data of Capital Medical University from May 1, 2010 to April 30, 2011. We analyzed the monthly clinic visiting rate of different diseases and disease distributions. We analyzed the average male and female college students in outpatient department, the proportion of students in different grades and different monthly clinic visiting rate of students coming from different locations, we also analyzed the distribution of different diseases in outpatients. Results For one month in average, much more female students than male students(22.9% and 13.2% of outpatients, P<0.05) visited the clinic; The students who came from other cities were much more than those from Beijing(26.3% and 15.3%, P<0.05); Significantly more students in grade two and three than students in grade one and four(24.9%, 25.6% and 12.6%, 12.6%, P<0.05) visits the clinic. For the distribution of different diseases, 48.2% had respiratory diseases, 14.3% had gastrointestinal disease, the other diseases only accounted for 37.5%. Conclusion Due to the special learning and living environment of medical students, we should develop appropriate measures to make appropriate medical and health education on medical students.

Phase 0 clinical trial: a new way to successful clinical trial for candidate drugs

ZHANG Jun;SONG Zi-hui;CAI Yong-ming;LIU Chang-xiao;ZHANG Zong-peng

2011, 32(4): 581-585. doi:10.3969/j.issn.1006-7795.2011.04.030

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Phase 0 trials are designed primarily to evaluate the pharmacodynamic and/or pharmacokinetic properties of selected investigational agents before initiating traditional phase I testing. In a phase 0 trial, the pharmacodynamic effects of an investigational agent are evaluated by a target or biomarker which is measured by an assay established and validated in preclinical models. The phase 0 trial is characterized by a small number of patients enrolled and limited dosing period with a non-therapy-intended dose range. Furthermore, phase 0 trials are classified into three types and conducted under the US Food and Drug Administration exploratory investigational new drug guidance with less preclinical toxicity data than usually required for traditional first-in-human studies. A well designed and executed phase 0 trial can overcome its limitations and has a great potential for improving the efficiency and success of subsequent trials, particularly those evaluating molecularly targeted antitumor candidates.

Present stats and problems in authorship with original articles published in selected medical journals

WANG Ying;LIU Li;LUAN Yi;KU Xue-fei;ZHANG Jun-min;HUANG Xiu-jiang;ZHAO Liang-jun;MIAO Li-juan

2011, 32(4): 586-589. doi:10.3969/j.issn.1006-7795.2011.04.031

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Objective To investigate the present status and problems in authorship in original articles published in medical journals. Methods A total of 9 429 authors of 3 156 original articles were selected from the following 6 journals: "Chinese Journal of Coal Industry Medicine", "Journal of Capital Medical University", "China Journal of Health Psychology", "Clinical Medicine of China", "Clinical Education of General Practice" and "Chinese Journal of Hospital Statistics" in 2010 were studied. A questionnaire survey was conducted among 2 468 authors of 988 papers each of which had more than 4 authors. Results The number of authors listed in each paper ranged from 1 to 16, and the average was 3. The number of grant at or above the provincial level was 1 030, of which 865 were signed, accounting for 84%; articles that had more than 4 authors had totally 2 468 authors, of whom only 1 086(44%) met the criteria of authorship; 475(89%) of the 532 first authors met the criteria of authorship; of the 43 corresponding authors, 38(88%) had author qualification. Conclusion Many of the authors of original articles published in the studied medical journals do not have clear awareness of "Copyright Law" and Academic Moral Ethics. Authors were indifferent to signature, unacquainted with concepts such as author, authorship and criteria for authorship.

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