The clinical data of 585 patients with connective tissue diseases (CTD) were retrospectively analyzed. Thyroid diseases were detected in 236 cases (40.3%) , in which the prevalence of hypothyroidism was the highest (47.9%) , followed by euthyroid sick syndrome (33.1%), Hashimoto thyroiditis (11.4%) and hyperthyroidism (5.1%) ect. Determination of thyroid function and autoantibodies to thyroid gland were essential in these patients.

Objective: To make a comparative observation on treating menopausal syndrome with auricular points sticking-pressure and acupuncture. Methods:The 86 cases were randomized into the group of auricular points sticking-pressure and acupuncture group, and then the comparative observation was made according to the modified Kupperman index. Results: The statistical analysis on the comparative observation showed there is no significant difference between the therapeutic effects of the two groups. Conclusion: Auricular points sticking-pressure and acupuncture are both effective for menopausal syndrome, and the auricular points sticking-pressure is worthy to spread with its easy operation.

Objective To study the application effect of health education guided by the theory of cross-theoretical model in the training of the first aid training in the basic units of the plateau. Methods Choosing 386 long-term presence of plateau soldiers, and divided them into the control group and the experimental group, the control group was educated by traditional method, in the experimental group, the health education was carried out with the trans-theoretical model, nursing experts gave them targeted intervention. The number of groups at each stage was analyzed in different stages, the same to examination results and pass rate. Results After 6 months of intervention, there was a significant improvement in the mastering of the battlefield Five emergency technology of the experimental group and the scores were better than those of the control group (P<0.05). Theoretical results showed that pass rate of the control group was 89.1%(172/193), the experimental group was 99.5%(192/193), the difference between the two groups was statistically significant (χ2=7.129, P=0.039). In the survey of technical operation, ventilation, hemostasis, bandaging, fixing and handling examination pass rate of the control group respectively were 88.1%(170/193), 89.1%(172/193), 89.1%(172/193), 88.1%(170/193), 88.6%(171/193), the experimental group were 99.5%(192/193), 97.9%(189/193), 99.5%(192/193), 96.9%(187/193), 97.4%(188/193), there were significant differences between the two groups (χ2=7.896-12.759, P=0.001-0.043). Conclusions Cross theoretical model has good effect on the application ofFiveemergency technology training in the high altitude troops battlefield, and can effectively improve the troops self- help and mutual aid capabilities, it is worthy of popularization and application.

Objective To observe the distribution of oligodexynucleotide (ODN)MT01 in main organ tissues of the rats at different time points and to discuss the regularity of the distribution of MT01 preliminarily. Methods 60 male Wistar rats were randomly divided into experimental group(n=30)and control group(n=30). The rats in experimental group was locally injected with Cy5 labeled MT01 in gingival mucosa,whereas the rats in control group were injected with MTO1.The samples of rat lung,liver spleen,kidney,heart,and brain tissues were collected at 15 min,1 h,4 h,8 h,16 h,1 d,2 d,3 d,4 d,and 5 d after injection,and the distribution of MT01 fluorescence was observed by laser scanning confocal microscope.The ratio of fluorescence positive cells indicated the amount of MT01 that had been taken up by different organs.Results No positive fluorescence cells were observed in control group.Whereas,in experimental group ,the positive fluorescence cells were detected in the tissue samples of lung,liver,spleen and kidney but not in the tissue samples of heart and brain.The positive fluorescence cells distributed focally in kidney tissue and presented primarily in the cytoplasm of renal tubular epithelial cells.The ratios of positive fluorescence cells changed regularly with time in liver, spleen and kidney tissues and the highest level was detected at 4,3 and 4 d after injection.No distinct regularity of the ratio of positive fluorescence cells was observed in lung tissue.Conclusion MT01 can be taken up by liver,spleen and lung tissue and primarily by kidney with regularity in distribution.

Objective To determine the effects of histone deacetylase inhibitor suberoylanilide hydroxamic acid(SAHA) on the cell proliferation and apoptosis of the human hepatic stellate cell line LX-2.The possible underlying mechanisms were also investigated.Methods The LX-2 cells were treated with SAHA in vitro.The morphology of LX-2 cells in different concentrations groups was observed by inverted microscope;the proliferation of LX-2 cells was measured by MTT assay;the Annexin V-FITC and PI staining was used to detect the apoptosis of LX-2 cells by flow cytometry and fluorescence microscope;the expression of α-SMA,collagen Ⅰ,acH3K9,acH3K14 and acH3K18 were detected by Western blot.Results The morphology change of LX-2 cells showed that SAHA inhibited the proliferation rate of LX-2 cells and in a dose dependent manner(P<0.05).The LX-2 cells were sensitive to SAHA along with time increasing,and in a time-dependent manner(P<0.05).Western blot showed that the expression levels of α-SMA and collagen-Ⅰ were significantly lower(P<0.05),on the contrary,the acetylation levels of acH3K9,acH3K14 and acH3K18 were significantly higher (P<0.05).Conclusions The increased acetylation of the histone acH3K9,acH3K14,acH3K18 and the lower expressed α-SMA and collagen-Ⅰ in LX-2 cells may be one of the mechanisms of SAHA.

AIM: To investigate the effect of a Chinese medicime, Dan-Shao-Hua-Xian capsule, on liver fibrosis induced by CCl_4 by observation of apoptosis and cell cycle variation in the liver cells. METHODS: Animal models were produced through eight-week treatment of the rats with CCl_4, alcohol and diet of high fat/low proteins, and then administration of Dan-Shao-Hua-Xian to the rats (1 g/kg) via stomach-tube-pouring for eight weeks was performed. Liver index, serum hyaluronic acid (HA) and glutamate pyruvate transaminase (ALT) were measured and hydroxyproline (Hyp) content in urine were determined. The extent of the liver fibrosis was observed under light microscope and apoptosis and cell cycle were also examined by cytometry between the two groups. RESULTS: Compared to the liver fibrosis group, the liver index, serum HA, ALT in the treatment group decreased, the development of liver fibrosis delayed, the urine Hyp and the number of apoptosed cells and the ratio of G_0/G_1 cells increased, as well as the S phase cells decreased, yet unable to return to normal. All those changes detected were statistically significant. CONCLUSIONS: The Dan-Shao-Hua-Xian is effective in treating the CCl_4-induced liver fibrosis in rats partly by virtue of inhibition of the growth of hepatic stellate cells and induction of apoptosis.

Objective To investigate the efficacy of sunitinib in the treatment of patients with imatinibresistant advanced gastrointestinal stromal tumor (GIST).Methods The clinical data of 45 patients with imatinib-resistant advanced GIST who received the treatment of sunitinib (37.5 mg/d) at the First Affiliated Hospital of Sun Yat-Sen University from March 2008 to June 2012 were retrospectively analyzed.The mutation of c-kit and platelet-derived growth factor receptor α (PDGFRα) was detected,and the efficacy of imatinib was assessed after the treatment for 3 months,and factors influencing the survival were analysed.The survival rate was calculated using the Kaplan-Meier method,survival analysis was done using the one-way analysis of variance,and multivariate analysis was done using the COX regression model.Results The median time of treatment with sunitinib for the 45 patients was 11.0 months (range,4-37 months).The complete remission rate,partial response rate,rate of stabilized condition and disease progression rate were 15.6％ (7/45),8.9％ (4/45),46.7％ (21/45) and 28.9％ (13/45) after the treatment with sunitinib for 3 months.All the patients with clinical (imaging) complete remission received surgery for metastatic lesions or B-ultrasound guided ablation for single liver metastasis before the treatment with sunitinib.The most common grade 3 or 4 adverse reactions of sunitinib were hand-foot syndrome and anemia.C-kit and PDGFRα mutational analysis were carried out.C-kit exon 9 mutation was detected in 9 patients,c-kit exon 11 mutation in 21 patients,and no mutation was detected in 12 patients.The median progression-free survival time was 8.0 months (range,4.1-11.9 months),and the median overall survival time was 25.0 months (range,13.4-36.6 months).The results of univariate analysis showed that the primary lesion sites and mutational status of primary lesions were factors influencing the progression-free survival and overall survival (x2=5.967,6.622 ; 7.965,8.765,P ＜ 0.05).The results of multivariate analysis showed that only the mutational status of c-kit of primary lesions was the independent factor influencing the progression-free survival and overall survival (Wald =6.540,7.205,P ＜ 0.05).The progression-free survival and overall survival of patients with c-kit exon 9 mutation and patients with no gene mutation were significantly longer than patients with c-kit exon 11 mutation (x2 =7.965,8.765,P ＜ 0.05).Conclusion Sunitinib with a dosage of 37.5 mg/d could effectively treat patients with imatinib-resistant advanced GIST.A better survival is observed in patients with c-kit exon 9 mutation or with no gene mutation when compared with patients with c-kit exon 11 mutation.

AIM:To investigate the changes of histone modifications during the activation of primarily cultured rat hepatic stellate cells ( HSCs) and the relationship between histone modification patterns andα-smooth muscle actin (α-SMA) expression, and to explore the roles of histone modifications in the activation of HSCs.METHODS:The rat HSCs were isolated by in situ perfusion of collagenase combined with density gradient centrifugation, cultured in vitro and identi-fied by immunofluorescence staining.The morphological features of the cells were observed under inverted microscope.The changes of desmin and α-SMA during the activation of HSCs were detected by immunofluorescence staining and Western blotting.The levels of histone 3 lysine 4 dimethylation (H3K4me2), histone 3 lysine 9 dimethylation (H3K9me2), his-tone 3 lysine 9 acetylation (acH3K9) and histone 4 lysine 12 acetylation (acH4K12) in quiescent HSCs and activated HSCs were determined by Western blotting.RESULTS: The morphology of HSCs shifted from a quiescent phenotype to highly activated myofibroblast during the culture.Immunofluorescence staining and Western blotting showed that the expres-sion levels of α-SMA and desmin were increased over time and reached maximum at 15 d.According to the results of cell morphology and immunofluorescence staining, the cells cultured for 24 h and 15 d were quiescent and activated HSCs, re-spectively.Compared with quiescent HSCs, there were higher H3K4me2 and lower H3K9me2, acH3K9 and acH4K12 modification levels in activated HSCs ( P<0.01 ) .CONCLUSION: Histone modifications show anomalous expression during the activation of primarily cultured rat HSCs.Histone modifications may contribute to the transdifferentiation of HSCs and the development of hepatic fibrosis.

Objective To characterize serum hepatitis B virus(HBV)full-length genome quasispecies and to investigate its ralationship with severe exacerbation of chronic hepatitis B(CHB).Methods HBV full-length genome was amplified and cloned from four treatment naive CHB patients and four treatment naive CSHB patients.Fourteen to sixteen clones per sample were selected,sequenced and analyzed by bioinformatics software.The measurement data was compared by independent-samples t test and count data was analyzed by x2 test. Results Totally 120 HBV fulllength genome sequences were obtained.All the patients had either genotype B or C virus monoinfection.One hundred percent clones(60/60)from CSHB patients showed mutations including G1896A,A1762T/G1764A(one patient even carried A1762T/G1764A/C1766T mutations),T1753C/G and start codon mutations in preS2,preS1,which were more common than those from CHB patients(46/60,76.7%;x2=15.85,P<0.01).The quasispecies complexity and diversity were higher in CSHB patients than CHB patients within full-length genome,S,X,P genes and reverse transcriptase region,but lower within C gene at both nucleotide and amino acid levels.But the difference were not statistically significant in all regions.Conclusion The mutation frequency and quasispeeies heterogeneity in HBV genome are higher in CSHB patients than in CHB patients,which may play a role in the severe exacerbation of CHB and needs further investigation in large scale studies.

Objective To characterize the profile and clinical significance of hepatitis B virus (HBV) quasispecies in patients infected with hepatitis B virus based on the sequence of reverse transcriptase (RT) region.Methods Fifty HBV infected treatment-naive patients were enrolled and divided into three groups,asymptomatic carriers (ASC) group (10 cases),chronic hepatitis B (CHB) group (30 cases) and liver cirrhosis (LC) group (10 cases).HBV genomes were extracted from serum samples.The sequence of RT region was amplified by polymerase chain reaction (PCR) and cloned into vectors.Fifteen to thirty clones per sample were selected,sequenced and analyzed by bioinformatics software.The mean values among groups were compared by analysis of variance.The median values among groups were compared by nonparametric statistics.The enumeration data were analyzed by x2 test.Results Totally 1221 HBV RT region nucleotide sequences were obtained (152from ASC patients,780 from CHB patients and 289 from LC patients).Genotype distribution showed no difference among three groups.However,the quasispecies complexity showed significant differences among the three groups,LC group ＞CHB group＞ ASC group (F=33.400,P＜0.05).The quasispecies diversity was LC group ＞CHB group＞ ASC group,and that of LC group was significantly different from the other two groups (F=18.070,P＜0.05),while there was no significant difference between CHB and ASC patients.Conclusions The HBV isolated from patients in immune clearance phase have higher variability than those isolated from patients in immune tolerance phase.The longer the infection persists and the more severe the disease is,the more variable HBV quasispecies are.