Objective To explore the mechanism of immune tolerance in chronic B hepatitis, the effects of HBV infection on the functions of macrophages and related mechanism of signal transduction by transfection in vitro were studied. Methods Peritoneal macrophages of mice were isolated regularly and cultured, transfected transiently with pcDNA3-HBV or pcDNA3 plasmid DNA and cultured under the stimulation of LPS. After 72 h, RT-PCR was performed to detect the expression of PreS1, TNF-? or IL-1? mRNA. To detect the expression of NF-?B RelA protein by FCM, and the level of nitric oxide in cultural supernatant was measured with Griess reaction. Results After being transfected with pcDNA3-HBV,peritoneal macrophages had the expression of PreS1 mRNA, but have lower level of TNF-?、IL-1? mRNA and transcriptional factor NF-?B, compared with pcDNA3-transfected control group; the level of nitric oxide in pcDNA3-HBV group was also decreased. Conclusions Transient transfection of pcDNA3-HBV could decrease the function of macrophages directly by inhibiting NF-?B activity and effector molecules production, which may be one of the mechanisms of immune tolerance in chronic B hepatitis.

BACKGROUND: Pharmacological researches demonstrate that Chinese crude drugs of invigorating the kidney contain flavonoids such as prepared rhizome of rehmannia and kudzuvine root, they possess phytoestrogenic effect, among them, isoflavone acts as a regulator of estrogen receptor.OBJECTIVE: To approach the effects of self-planned Bushen Huoxue Fang (BSHXF) on the serum estradiol and estrogen receptor α mRNA expression in pituitary gland in rats with collagen-induced arthritis. DESIGN, TIME AND SETTING: Randomized grouping controlled animal experiment was performed in the central laboratory of Bethune Intemational Peace Hospital of Chinese PLA between May 2006 and May 2008. MATERIALS: Sixty female and healthy Wistar rats were randomly divided into sham operated group (n=10) and operation group (n=50). Operation group was reproduced collagen-induced arthritis model at 2 weeks following ovariectomy. The rats of arthritic index ≥ 5 were assigned into model group, BSHXF group, methotrexate (MTX) group, and MTX + BSHXF group, each group included 10 rats. The Chinese crude drugs were bought from LERENTANG drugstore in Shijiazhuang, including rhizoma drynariae, safflower, prepared rhizome of rehmannia, Herba Epimedii, danshen root, kudzuvine root and so on. After routine decoction, centrifugalization, rogue and condensing 1.3 kg/L decoction, the solution was sealed up, divided pack and reserved at 4℃.METHODS: Sham operated group and model group were intragastrically administered with normal saline 2 mL per day; BSHXF group with BSHXF decoction 2 mL per day; MTX group with MTX 2.6 mg/kg (2 mL) once a week, and normal saline 2 mL per day at other time points; MTX + BSHXF group with MTX 2.6 mg/kg (2 mL) once a week and BSHXF juice 2 mL per day. Each group was given 6 weeks.MAIN OUTCOME MEASURES: After blooding by abdominalis aorta, magnetic-split enzyme linked immunosorbent assay was applied to detect the level of serum estradiol. Reverse transcription-polymerase chain reaction detected the expression of estrogen receptor a mRNA in rat pituitary gland.RESULTS: Compared with model group, the expression of estrogen receptor a mRNA in pituitary gland increased in the MTX group, but there was not statistically significant. BSHXF + MTX group showed a significantly increased expression (P < 0.01). The expression of estrogen receptor α mRNA in BSHXF group was higher than MTX group, but lower than BSHXF + MTX group (P < 0.05). No significant difference existed between BSHXF group, BSHXF + MTX group and sham operation group. Compared with sham operation group, there were statistically significant decrease of estradiol in the model group. Compared with model group, the estradiol levels were shown to increase in three treatment groups. Among them, MTX group had slightly slower level with no significant difference (P > 0.05), while BSHXF + MTX group had the most obvious increase, which was close to normal level (P < 0.01). There was a significantly positive correlation between estradiol and the expression of estrogen receptor o mRNA (r=0.483, P < 0.05).CONCLUSION: Serf-planned BSHXF possessing oestrogenic hormonelike role can elevate the level of serum estradiol and upregulate the expression of estrogen receptor a mRNA in pituitary gland.

Objective To study the expression of HER-3 in breast cancer patients and analyze its relationship with the clinical pathology and prognosis of breast cancer,ER,PR and HER-2.Methods Data of 139 female breast cancer patients undergoing operations at the Second Hospital of Dalian Medical University from Jan.2006 to Oct.2006 were collected.These 139 patients didn't have preoperative chemotherapy or radiation therapy,but had postoperative endocrine therapies.Immunohistochemisty was used to detect the expression of HER-3 in the 139 patients.Data like the expression of ER,PR and HER-2,histological grading,tumor size and lymph node metastasis were obtained from their medical records.The relationship between indicators and breast cancer was analyzed.Results ①The positive expression rate of HER-3,HER-2,ER,and PR was 30.2％ (42/139),42.5％ (59/139),66.9％ (93/139),and 59.7％ (83/139) respectively.(②)The statistical items had no relation with patients' age(P ＞0.05).③HER-3 expression was correlated with tumor sizes,lymph node metastasis,histological grading,5 year survival rate and HER-2 expression(P ＜ 0.05),and had no relation with ER or PR(P ＞0.05).④)The combined expression of HER-3 & HER-2 was correlated with tumor sizes,axillary lymph node metastasis,histological grading,ER and PR.(P ＜ 0.05).Conclusions HER-3 detection is of great significance for the diagnosis and prognosis of breast cancer.The combined examination of HER-3,HER-2,ER,and PR in breast cancer tissues is of clinical significance for early diagnosis/treatment,medication and prognosis of the tumor,as well as for new drug research and development.

AIM: To study the effect of cholecystokinin octapeptide(CCK-8) on lipopolysaccharide (LPS)-stimulated pulmonary interstitial macrophage(IM) in vitro. METHODS: Pulmonary IM were isolated and cultured in the presence of LPS, CCK-8, proglumide(the antagonist of CCK receptors) and vehicle alone or together. The expression of mCD14 protein was assayed by flow cytometry, and sCD14 in the supernatant was analyzed semi-quantitatively by Western blot, and TNF-? in the supernatant was detected with ELISA. RESULTS: CCK-8, at concentrations from 10 -7 mol/L to 10 -6 mol/L inhibited significantly the expression of mCD14, the release of sCD14 and TNF-? to the supernatant up-regulated by LPS(1 mg/L). The effect of CCK-8 was inhibited by proglumide. CONCLUSION: CCK-8 modulated negatively several functions of LPS-stimulated pulmonary IM through CCK receptors, which may be one of the mechanisms for CCK-8 to alleviate the inflammation in lung tissues during endotoxemia.

AIM: To explore the effect of TNF-related apoptosis inducing ligand (TRAIL), a new apoptotic inducing molecule on the biological activity of hepatocarcinoma cell line. METHODS: The expression of membrane binding TRAIL on HepG2 cells was detected by immuno-cytochemistry. Quantity of secretory TRAIL was assayed by ELISA method. The cytotoxicity and apoptosis induced by TRAIL was detected by MTT and TUNEL method, respectively. The telomerase activity of HepG2 cells was detected by TRAP-PCR assay kit. The expression of hTERT, the catalytic subunit of telomerase, was detected by FCM. RESULTS: TRAIL was constitutively expressed on the membrane of HepG2 cell line. Soluble TRAIL was also expressed to a certain degree. Cytotoxicity assay showed that TRAIL significantly inhibited the growth of hepatocarcinoma cells. TUNEL assay indicated that TRAIL induced apoptosis in hepatocarcinoma cells. Detection of telomerase activity showed that TRAIL inhibited telomerase activity and the expression of telomerase catalytic subunit. CONCLUSION: TRAIL is an effective molecule to inhibit the growth of hepatocarcinoma through multiple pathways, such as inducing apoptosis and inhibiting the activity of telomerase.

Objective To construct eukaryotic expression vector of small hairpin RNA vector targeting hepatitis B virus core protein in HepG2.2.15 cells,and to verify the interfering effect.Methods To design and synthesize small interfering RNA sequence against HBc gene (ayw subtype) which locates between 2147 bp to 2165 bp in the HBV genome.The siRNA fragment were inserted into pSilencer3.1-H1 neo vector.The recombinants were transformed into E.coli DHSα successfully.The recombinant plasmids were identified by Eco RI enzyme digestion and sequenced.Finally,The HepG2.2.15 cells were transfected with the vector by using lipofectamine method to identify its interfering effect.Results The eukaryotic expression vector containing small hairpin RNA targeting HBV core protein was constructed successfully and named as pshRNA-HBc.The recombinant plasmids were identified by SalI or EcoRI enzyme digestion and sequenced.The silencing effect was detected by using revere transcription polymerase chain reaction (RTPCR) and the secretion of HBsAg and HBeAg in the supernatant of HepG2.2.15 cells.The result showed that the interfering effect was specific and efficient.Conclusion The small interfering RNA targeting HBV core protein in HepG2.2.15 cells was constructed successfully,which may be useful for studying the biological functions of HBc in the future.

Objective To analyze the differences of clinical manifestations and organ damage between patients with severe fever with thrombocytopenia syndrome(SFTS)and patients with tsutsugamushi disease,and to investigate the prognostic factors of SFTS.Methods The research was performed on 49 patients with SFTS and 16 patients with tsutsugamushi disease who visited the First Affiliated Hospital of Anhui Medical University from October 2014 to June 2017.The general information of patients including region,age,gender and clinical manifestations were evaluated.Blood routine,liver and kidney function,myocardial enzyme levels,lipase,amylase,electrolytes,C-reactive protein,procalcitonin,prothrombin time(PT)and activated partial thromboplastin time(APTT)were continuously monitored during the course of disease.T test was used for continuous variables of normal distribution,and non-parametric test was used for variables of non-normal distribution.Chi-square test was used for categorical variables.Results The mean age of SFTS patients was 62.1±15.5(ranging from 17 to 87 years)and the mean age of tsutsugamushi patients was 56.1±9.2(ranging from 47 to 73 years).There was no significant difference between the two groups(t=1.47,P=0.147).There were 25 males(51%)in SFTS patients and 8 males(50%)in tsutsugamushi disease patients.There was no significant difference between the two groups(x2=0.005,P=0.943).The incidences of headache,vomiting,superficial lymphadenectasis,disturbance of consciousness,proteinuria,hematuria,pulmonary infection,multiple organ dysfunction and acute pancreatitis in SFTS patients were all significantly higher than those in tsutsugamushi disease patients(x2=8.82,4.38,8.71,11.17,7.88,5.56,4.35,9.43,and 8.13,respectively,P <0.05 or 0.01).The counts of leukocytes(Z=2.73),neutrophils(Z=2.46),lymphocytes(Z=3.15),platelets(Z=4.25),albumin(Z=2.65)and sodium ion(t=2.10)in SFTS patients were all significantly lower than those in patients with tsutsugamushi disease(P <0.05 or 0.01).The levels of aspartate aminotransferase(Z=2.94),lactate dehydrogenase(Z=3.42),creatine kinase(CK)(Z=2.88),amylase(Z=2.11),lipase(Z=2.82),creatinine(Z=2.07)and urea nitrogen(Z=2.50)in fatal SFTS patients were all significantly higher than those in patients with tsutsugamushi disease(P <0.05 or 0.01).Among 49 SFTS patients,16 patients died and 33 patients recovered finally.The age(t=3.33),platelet count(Z=2.55),alanine aminotransferase(ALT)(Z=2.10),aspartate aminotransferase(AST)(Z=2.22),lactate dehydrogenase(Z=2.26),CK(Z=3.50),CK-MB(Z=3.10),creatinine(Z=2.17),urea nitrogen(Z=2.36),and sodium(t=2.65)between the two subgroups had significant differences(P <0.05 or 0.01).Conclusions SFTS is more severe and has high mortality,while tsutsugamushi disease has a better prognosis.Early differential diagnosis and early rational treatment are important to reduce the mortality of patients with SFTS.

A total of 207 general practitioners who participated in the "Beijing-Hong Kong Community Health Service Training Demonstration Center Project" training from 2013 to 2017 were selected for this questionnaire survey. A self-designed questionnaire was used, which contained the basic information of the trainees, the training purpose, the shortcomings of training, the satisfaction and suggestions for training. Total 192 valid questionnaires were retrieved with a recovery rate of 92.8%. The in-depth personal interviews were conducted among 8 trainees who served as key trainers in community health service centers and 4 managers from their institutions, to investigate the subsequent application of the learning results after they returning to work. The survey showed that the reasons of participating in training were knowledge updates (165, 85.94%), competence requirements (128, 66.67%), and due to administration decision (127, 66.15%). The highest comprehensive scores of training needs were improvement of general practice thinking ability (4.63), followed by clinical diagnosis and treatment ability (4.44), and general practice skills (3.97). For teaching contents, participants were most interested in diagnosis ability (4.03) and clinical operative techniques (3.20). For the training form, the participants more favored the case discussion method (80.73%, 155/192), clinical practice (52.08%, 100/192) and community practice (38.02%,73/192). The trainees were highly satisfied with the overall training organization, training time, training form, and training teachers of the Beijing-Hong Kong training project, and had a good grasp of the training content. Half of trainees (97, 50.52%) thought that the main shortcomings of training was the discrepancy between training content and practical work. Interviewees showed that after the training, the team management, communication skills and disease management ability for some minor specialties had been significantly improved; the enthusiasm for scientific research had been stimulated; they had a certain managerial thinking; and were able to apply what they leant in community health service. In summary, the training project has reached the initial training goals to promote the comprehensive ability of trainees.

Objective:To compare epidemic characteristics between severe acute respiratory syndrome (SARS) and coronavirus disease 2019 (COVID-19).Methods:The general information, including epidemiological and clinical data of the confirmed cases during the epidemic period of the two infectious diseases was collected. The data of SARS in Guangzhou was derived from the technical files of Guangzhou Center for Disease Control and Prevention (GZCDC), including the statistical report and brief report of the epidemic situation (from January 2 to May 11, 2003);The data of COVID-19 in Wuhan was derived from the epidemic data published by Wuhan health and Health Committee's official website and other publicly reported documents (from December 9, 2019 to March 11, 2020). Descriptive analysis was used for a comparativeanalysis of the time and age characteristics, the number of cases, basic reproduction number (R0), proportion of medical staff in confirmed cases (%), crude mortality, etc.Results:A total of 1 072 cases of SARS in Guangzhou were included in the study. The incidence ratio of male to female was 1∶1.26. 43 cases of death were reported with a mortality rate of 4.01%. The median age was 36 years old. The proportion of medical staff in the early stage of the epidemic was 29.04% (88 cases). As to COVID-19 in Wuhan, a total of 49 978 cases were included, The incidence ratio of male to female was 1.04∶1. The 2 423 cases of death were reported with a mortality rate of 4.85%. The median age was 56 years old. The proportion of medical staff in the early stage of the epidemic was 30.43% (42 cases).Conclusion:The COVID-19 in Wuhan has the characteristics of high incidence and wide population. However, the epidemic situation is falling rapidly, and the prevention and control strategy needs to be adjusted timely. The prevention and control of nosocomial infection should be addressed in future.

Objective:To compare epidemic characteristics between severe acute respiratory syndrome (SARS) and coronavirus disease 2019 (COVID-19).Methods:The general information, including epidemiological and clinical data of the confirmed cases during the epidemic period of the two infectious diseases was collected. The data of SARS in Guangzhou was derived from the technical files of Guangzhou Center for Disease Control and Prevention (GZCDC), including the statistical report and brief report of the epidemic situation (from January 2 to May 11, 2003);The data of COVID-19 in Wuhan was derived from the epidemic data published by Wuhan health and Health Committee's official website and other publicly reported documents (from December 9, 2019 to March 11, 2020). Descriptive analysis was used for a comparativeanalysis of the time and age characteristics, the number of cases, basic reproduction number (R0), proportion of medical staff in confirmed cases (%), crude mortality, etc.Results:A total of 1 072 cases of SARS in Guangzhou were included in the study. The incidence ratio of male to female was 1∶1.26. 43 cases of death were reported with a mortality rate of 4.01%. The median age was 36 years old. The proportion of medical staff in the early stage of the epidemic was 29.04% (88 cases). As to COVID-19 in Wuhan, a total of 49 978 cases were included, The incidence ratio of male to female was 1.04∶1. The 2 423 cases of death were reported with a mortality rate of 4.85%. The median age was 56 years old. The proportion of medical staff in the early stage of the epidemic was 30.43% (42 cases).Conclusion:The COVID-19 in Wuhan has the characteristics of high incidence and wide population. However, the epidemic situation is falling rapidly, and the prevention and control strategy needs to be adjusted timely. The prevention and control of nosocomial infection should be addressed in future.