Objective:To prepare and purify recombinant human zona pellucida 3 protein,and to study its immunologic activity.Methods:The E.coli BL21 containing recombinant plasmid pGEX4T-1/ huZP3 was induced to express GST-fusion protein by IPTG.After a series of purification procedure,the purified protein was analyzed by SDS-PAGE.After the mice immunized with rhuZP3,the antibody responses against rhuZP3 were detected by ELISA.Results:The soluble fusion protein was expressed,and purity of rhuZP3 was 95%.Moreover, purity of rhuZP3 could be recognized by anti-human ZP3 in ELISA.Conclusion:The rhuZP3 is obtained through the preparation of prokaryotic expression system and anti-rhuZP3 antibody has immunological activity.

Objective To observe the influence of Fuzheng Chuyi Granules on the mice infected with influenza virus and to observe its antibiotic and antivirus actions in vitro.Methods The death rate of mice infected with influenza virus in each group was compared and antimicrobial activity of Fuzheng Chuyi Granules was observed in vitro.Hep-2 passage cells were subcultured to observe the protective effect of the maximal non-toxic dosage(TD0),the infective dose of 50% tissue culture(TCID50) and other different dosage of Fuzheng Chuyi Granules on cells infected virus.Results Compared with that in negative control group,the death rate of infected mice was lower in Fuzheng Chuyi Granules groups.Fuzheng Chuyi Granules had antibiotic action on bacteria in vitro,particularly on streptococcus pneumoniae.Meanwhile,Fuzheng Chuyi Granules could alleviate cytopathy induced by virus.Conclusion Fuzheng Chuyi Granules have a certain protective effect on the animal infected with virus,and also have antibiotic and antivirus actions.

Objective:To investigate the anti-inflammation,immune regulation and anti-stress effect of cartialgenous collagen zymolyte.Methods:The anti-inflammatory effect was observed by carrageenan-induced foot swelling test,the immune effect was observed by delayed-type hypersensitivity (DTH) induced by 2,4-dinitrochlorobenzene and the anti-stress effect was observed by hypoxia tolerance and fatigue tolerance. Results:The cartialgenous collagen zymolyte can inhibit the carrageenan-induced foot swelling and delayed-type hypersensitivity (DTH) induced by 2,4-dinitrochlorobenzene,and improve the macrophagocyte phagocytosis function,and enhance the body,s non-specific resistance to fatigue.Conclusion:Cartialgenous collagen zymolyte appeared obvious immune regulation,anti-inflammation and anti-stress effect.

BACKGROUND: Cardiac stem cells can differentiate into cardiomyocytes in vitro under induction of pilose antlerpolypeptides, which provides a new therapeutic idea for myocardial injury.OBJECTIVE: To explore the effect of pilose antler polypeptides on the apoptosis rate and membrane stability ofmitochondria in cardiac stem cells.METHODS: We chose healthy male Wistar rats born 2 days to extract cardiac stem cells. The culture dish was used asthe experimental unit, and extracted cells were divided into the following four groups (n=12). Blank control group: Thesame amount of buffer was added for induction; 5-azacytidine group: induced with 5-azacytidine (3 μmol/L); pilose antlerpolypeptides group: induced with pilose antler polypeptides (800 mg/L); combined group: induced with pilose antlerpolypeptides (800 mg/L) and 5-azacytidine (3 μmol/L). After 48 hours induction, apoptosis rate of cardiac stem cells ineach group was detected with flow cytometry. The membrane potential of mitochondria in cardiac stem cells wasdetected with immunofluorescence. The expression level of Nkx 2.5, GATA4, ATF-2, and MEF-2C in cardiac stem cellswas detected using western blot assay.RESULTS AND CONCLUSION: There were small, round and bright cells after 2 weeks culture, and cell colonies ofcardiac stem cells formed. The apoptosis rate of cardiac stem cells in the 5-azacytidine group increased significantly (P <0.05) and its membrane potential decreased significantly compared with the blank control group (P < 0.05). Theapoptosis rate of cardiac stem cells in the pilose antler polypeptides group and combined group decreased significantlycompared with the 5-azacytidine group (P < 0.05, P < 0.01), and their membrane potential increased significantly (P <0.05, P < 0.01). The results of western blot showed that the expression level of Nkx2.5 increased significantly in thepilose antler polypeptides group and combined group compared with the 5-azacytidine group (P < 0.05), while theexpression levels of GATA4 and ATF-2 in each group were low and there were no significant differences among groups(P > 0.05). The expression level of MEF-2C in each group was at a middle level, and there were no significantdifferences among groups (P > 0.05). To conclude, our experimental findings indicate that pilose antler polypeptidescould decrease the apoptosis rate and improve membrane stability of mitochondria in cardiac stem cells. Themechanism may be related to the increased expression of Nkx 2.5, but whether the mechanism is related to GATA4,ATF-2 and MEF-2C needs to be further studied.

Objective To investigate the effect of body weight on the induction of mild hypothermia in a rabbit model of asphyxia cardiac arrest. Methods Twenty-four rabbits were randomized into two groups: the ice bag group and the intravenous 4℃ saline group. Cardiac arrest was induced and after 3 minutes of cardiac arrest, cardiopulmonary resuscitation was begun. Simultaneously, mild hypothermia was induced by putting an ice bag over the abdomen or infusion of 4℃ saline via an ear vein. A 2℃ decrease of rectal temperature was considered as the completion of hypothermia induction. Induction times were recorded, compared, and analyzed with respect to body weight. Results All rabbits had restoration of spontaneous circulation (ROSC) and ROSC lasted during the experiment. Induction time in the ice bag group was significantly shorter than that in the intravenous 4℃ saline group (22.8±4.7 min VS 42.5±4.0 min, P< 0.001). Induction time significantly correlated with body weight in the ice bag group (Pearson Correlation: r = 0.725, P = 0.029), but not in the intravenous 4℃ saline group (Pearson Correlation: P = 0.418). Conclusions In a rabbit model, induction of mild hypothermia with an ice bag is faster than with intravenous 4℃ saline; induction time positively correlates with body weight when an ice bag is used, but not when intravenous 4℃ saline used. The effect of body weight should be considered when choosing an appropriate method to achieve early induction of mild hypothermia.

Objectives:To validate the Chinese version of the Ureteral Stent Symptoms Questionnaire(Chinese-USSQ) in patients with an indwelling ureteral stent.Methods:The original USSQ was translated into Chinese and linguistically validated following the cross-cultural adaptation of health-related quality of life measures.A total of 83 patients (cases) with indwelling ureteral stent and 90 healthy subjects (controls) were asked to complete the Chinese-USSQ as well as European Quality of Life Visual Analogue scale(EQ-VAS)(for both genders), the International Prostate Symptom Scale (IPSS) (for male), and Urogenital Distress Inventory-6 (for female). Patients were evaluated at weeks 1 and 4 after stent placement and at week 4 after removal. The psychometric properties of the questionnaire were analyzed.Results:The Chinese version of USSQ include 43 items, which addressed various domains of health(6 sections) covering urinary symptoms, pain, general health, work performance and sexual matters with additional problems. A total of 78 patients(45 males and 33 females) and 90 controls (41 males and 49 females)were included for analysis. Internal consistencies (Cronbach′s α coefficients: 0.60-0.78) and test-retest reliability (Spearman correlation coefficient: 0.69-0.91) were satisfactory for urinary symptom, body pain, general health, and work performance domains. Most USSQ domains showed moderate correlations with each other. The convergent validity determined by correlation between other instruments and corresponding USSQ domain was satisfactory. At week 4 it was moderate for the urinary symptoms index compared to the IPSS in men(Spearman correlation coefficient>0.60), for the urinalry symptoms index compared to the UDI-6 score in women(Spearman correlation coefficient 0.52).Sensitivity to change and discriminant validity were also good in most domains ( P<0.01). Only a small proportion of the study population had an active sexual life with the stent in situ, which limited its analysis. Only 1(1.3%) and 6(7.7%) patients had an active sex life at week 1 and 4 after stent placement; meanwhile, 34(43.6%) patents were sexually active at week 4 after stent removal. Conclusion:The Chinese version of the USSQ is a reliable and valid instrument that can be used for Chinese patients with a indwelling ureteral stent in the clinical and research settings.

The objective was to evaluate the preventive and therapeutic effects of the collagen hydrolysate extracted from Sika deer velvet (CSDV) on osteoporosis rats induced by retinoicacid. Histomorphometric indices and serum biochemical parameters were measured in osteoporosis rats treated with/without antler collagen and in sham-operated rats. Our results were as follows: compared with the osteoporosis group, significant elevation in the levels of bone mineral density (BMD), Ca, P and static histomorphometric indexes and biomechanical properties, but reduction in the level of serum alkaline phosphatase (ALP) were observed in antler collagen-treated groups. However, the above function with the collagenase solution velvet material varied with the different doses. In conclusion, the extracted collagen is found to play a role in the prevention and treatment of osteoporosis rats by retinoic acid.
Animals ; Bone Density ; drug effects ; Collagen ; metabolism ; therapeutic use ; Female ; Humans ; Male ; Osteoporosis ; prevention & control ; Osteoporosis, Postmenopausal ; Rats ; Rats, Wistar ; Tretinoin ; therapeutic use

Objective To analyze the distribution of common chromosomal karyotypes of patients with Turner syndrome (TS), and to explore the correlation between the age and height standard deviation scores (HSDS) on diagnosis.Methods Retrospective investigation was performed for the data of age and HSDS on diagnosis in 273 TS girls(≤ 18.0 years old)diagnosed by chromosomal karyotypes.The main statistical methods were analyzed with t-test and Pearson correlation test by using the SPSS 18.0 statistical software.Results (1) There were 4 kinds of common chromosomal karyotypes in the TS :45, X (87/273 cases,31.9％),46, X, i (Xq) (43/273 cases, 15.7％) ,45, X/46, X, i (Xq) (36/273 cases, 13.2％) and 45, X/46, XX (23/273 cases, 8.4％), respectively, the adolescent TS all had delayed puberty.For the cases with 45, X karyotypes ,3 cases presented mental retardation and 2 cases with organs deformity.(2)The patients with 45 ,X/46,X,i(Xq) karyotypes or with 46,X,i(Xq) karyotypes had the maximum(12.56 age) or the minimum(9.70 age) mean age on diagnosis, respectively, there was a significant difference between 2 groups (t =3.019, P =0.004).The maximum deviation from normal height was found in the patients with karyotypes of 46, X,i (Xq) (HSDS =-4.04), and the minimum deviation was in the patients with karyotypes of 45,X/46, XX (HSDS =-3.16), and there was a significant difference between 2 groups (t =-2.95, P =0.004).(3) More than 75.7％ of TS patients was diagnosed when their heights deviated above 3 SD,and their mean age on diagnosis was 12.10 age,which was 3 years later than those patients within 2 SD.(4) There was a significant negative correlation between the age and HSDS on diagnosis in the groups of common chromosomal karyotypes[45,X、46,X,i(Xq) and 45,X/46,XX] (r =-0.551,-0.560,-0.622,all P ＜ 0.01), except for the group with the 45, X/46, X, i (Xq).Conclusions (1) In this study, the consti-tuent ratios of these 4 common chromosomal karyotypes were different from those in Europe and America's.(2)Patients with 45 ,X may have more severe symptoms than others.(3)The mean age on diagnosis was at least 3.0 years earlier when considered HSDS below-2.00 as an indicator for chromosomal karyotype screening,which would facilitate earlier diagnosis.

Background: Early diagnosis and staging of liver fibrosis are important for the prognosis and evaluating the survival of patients.Aims: To systematically assess the diagnostic value of transient elastography (TE) for staging of liver fibrosis in patients with chronic liver disease.Methods: PubMed,Embase,Cochrane Library,CNKI,Wanfang and VIP from Jan.2001 to Dec.2015 were retrieved to collect the articles with staging of liver fibrosis in patients with chronic liver disease by TE.Data extraction was conducted.Article quality was evaluated by quality assessment of diagnostic accuracy studies 2 (QUADAS2).Meta-analysis was conducted by Stata 12.0 software.Results: Twenty articles involving 5 748 patients were included.Meta-analysis showed that the combined sensitivity,specificity and AUC of TE for diagnosing significant fibrosis (≥F2) were 0.78 (95％ CI: 0.73-0.82),0.85 (95％ CI: 0.80-0.88) and 0.88 (95％ CI: 0.85-0.91),respectively.The combined sensitivity,specificity and AUC for advanced fibrosis (≥F3) were 0.89 (95％ CI: 0.86-0.91),0.88 (95％ CI: 0.85-0.91) and 0.94 (95％ CI: 0.92-0.96),respectively.The combined sensitivity,specificity and AUC for cirrhosis (F4) were 0.91 (95％ CI: 0.86-0.95),0.89 (95％ CI: 0.87-0.92) and 0.95 (95％ CI: 0.93-0.97),respectively.Conclusions: TE technique has a good diagnostic value in assessing significant fibrosis,advanced fibrosis and cirrhosis in patients with chronic liver disease,especially for advanced fibrosis and cirrhosis.