To investigate the quality of prepared pieces of Stephania tetrandra S.the content of total alkaloids and tetrandrine of the products prepared by 10 different prepared pieces of Chinese medicine factories has been determined. In conclusion, it is practicable to prescribe the content of total alkaloids and tetrandrine for 3% and 1%, respectively.

BACKGROUND:To improve the survival rate of transplanted hematopoietic stem cells, dynamic monitoring of plasma content of human cytomegalovirus (HCMV) and rapid screening of early active HCMV infection in hematopoietic stem celltranplantation recipients, thus to guide the clinical medication, is preferred. OBJECTIVE:To evaluate the usefulness of fluorescence quantitative PCR assay for early detection of HCMV activation after hematopoietic stem celltransplantation. METHODS:Real-time fluorescence quantitative PCR assay was applied for HCMV monitoring in 656 blood samples from 41 hematopoietic stem celltranplantation recipients and 60 control blood samples. RESULTS AND CONCLUSION:In 656 blood samples, 96 samples were positive, and the HCMV DNA copies ranged from 5.0×102 to 1.0×107 IU/mL. Timely initiation of therapy resulted in the rapid clearance of DNA-viraemia but it recurrenced in short time by drug-resistent. Two cases from 12 postive recipients died from HCMV infection. The quantitative detection of HCMV DNA by real-time PCR is a rapid method for monitoring HCMV infection and the early diagnosis in patients after hematopoietic stem celltransplantation.

Objective To explore the safety and feasibility of blocking blood flow in interstitial tubal pregnancy treated with laparoscopic opening-taking embryo operation.Methods The clinical data of 98 patients with lump interstitial tubal pregnancies (requesting reserve procreate function) from January 2006 to December 2013 were chosen.Among them,56 patients were in study group (January 2010-December 2013) and 42 patients were in control group (January 2006-December 2009).All patients were treated with opening-taking embryo by laparoscopic operation.In study group,we first blocked the uterine artery and ovarian artery blood supply of pregnancy lump,secondly opened pregnancy lump and stripped gestation sac with hydraulic pressure separation during operation.Whereas,in control group,we opened pregnancy lump and taken out pregnancy tissues according to convention method without blocking blood flow.Operation success rate,operation blood volume,operation time,persistent ectopic pregnancy (PEP) happening rate,fallopian tube unobstructed information,and pregnancy information after operation were compared between two groups.Results In study group,operation success rate was 96.4％,which was significantly higher than that in control group (61.9％) (P ＜0.01) ; operation blood volume was[(20.7 ± 10.4)ml],which was significantly less than that in control group [(60.7 ± 18.4) ml] (P ＜ 0.01) ; operation time [(46.6 ±14.2) min] was significantly shorter than that in control group [(66.5 ± 19.4) min] (P ＜ 0.01) ; there was no PEP in study group,while there were 5 PEPs (11.9％) in control group.Fallopian tube unobstructed rate after operation in study group (76.9％) was significantly higher than that in control group (41.7％) (P ＜ 0.05).Conclusions Application of blocking blood flow in opening-taking embryo by laparoscopic operation on lump interstitial tubal pregnancies is safe and effective.

Objective To examine the expression of stromal cell-derived factor-1 (SDF-1) in a rat model of retinal ischemia-reperfusion injury (RIRI),and investigate the protective effect of 17β-Estradiol (E2) on RIRI and explore the mechanism.Methods The RIRI model was established in Sprague-Dawley rats by increasing the intraocular pressure.Relative expression levels of SDF-1 mRNA and protein in the retina at 6 hours,12 hours and 24 hours following reperfusion was determined by RT-PCR and Western blot,respectively.E2 was administered to investigate the effects of estrogen on SDF-1 expression,and the estrogen receptor antagonist ICI 182-780 was administered to investigate the effect of estrogen receptor on the expression of SDF-1.Results SDF-1 expression in RIRI 6 hours group,12 hours group and 24 hours group was increased compared with normal control group (all P ＜ 0.05),with maximum expression at RIRI 12 hours group.As expected,pretreatment of RIRI rats with E2 had a protection on RIRI retina;SDF-1 expression was increased in RIRI + E2 group compared with IR control group and RIRI + vehicle group (all P ＜ 0.05).RIRI + E2 + ICI 182-780 group could decrease SDF-1 expression compared with RIRI + E2 group(all P ＜ 0.05).Conclusion E2 offers protection against RIRI by inducing an up-regulation in SDF-1 expression through activation of the estrogen receptor.

Objective To evaluate whether the serum uric acid (SUA) levels are related to development of prediabetes. Methods This was a 6-year cohort study, subjects were recruited from Tianjin Medical University General Hospital Health Management Center. A prospective assessment (n=30 910) was performed. Subjects without a history of prediabetes were followed up for 6 years (with a median follow-up of 2.7 person-years). All relevant variables including SUA concentrations, and fasting plasma glucose were assessed at baseline and yearly during the follow-up. Prediabetes was defined in accordance with the American Diabetes Association criteria of 2014. Multivariate-adjusted Cox proportional hazards regression models were fitted to assess relationships between the quintiles of baseline SUA and the incidence of prediabetes. Results The incidence of prediabetes was 55 per 1 000 person-years. In comparison with subjects in the lowest quintile, the odds ratios and hazard ratio (95%confidence interval) of higher quintile of SUA were 1.04 (0.93, 1.15), 1.07 (0.95, 1.19), 1.13 (1.01, 1.28) and 1.15 (1.02, 1.30) (P for trend=0.01), respectively, after adjusting for potential confounders. Conclusion SUA levels predicted an increased risk of prediabetes in adults.

Objective To optimize the culture method for rheumatoid arthritis fibroblast-like synoviocytes in vitro,and observe the effect of Dishevelled (Dvl) 2 on vascular endothelial growth factor (VEGF) in rheumatoid arthritis fibroblast-like synoviocytes (RA-FLS).Methods Synovium from RA patients who underwent knee arthroplasties were cut into small piece,and RA-FLS were isolated and cultured in vitro using tissue block method.Dvl 2 lentivirus overexpressing plasmid was constructed and transfected into RAFLS.Q-polymerase chain reaction (PCR) and enzyme linked immunosorbent assay (ELISA) were used to detect the mRNA and protein expression levels of VEGF.Then we used 10 ng/ml tumor necrosis factor (TNF)-α recombinant protein to stimulate the transfected RA-FLS.24 h after stimulation,mRNA and protein expression of VEGF were detected again.Student's t test was used for two group analyses.Results RA-FLS was successfully isolated and cultured in vitro.The multiplicity of infection was 30 and was in conjunction with appropriate concentration of polybrene to promote transfection.Transfection efficiency could meet the test requirements.The mRNA of Dvl 2 increased for 79-fold than the control group.Compared with the control group,Dvl 2 could mildly inhibit RA-FLS secretion of VEGF.After TNF-α stimulation,Dvl 2 could significantly inhibit the VEGF's mRNA (2.15±0.10,2.92±0.47 fold,t=-3.924,P=0.003) and protein [(285±100) pg/ml,(155±61) pg/ml,t=-2.714,P=0.022] expression compared with the control group.Conclusion Dvl 2 can inhibit the effect of TNF-α induced secretion of VEGF in RA-FLS.The specific mechanism needs further study.

Objective This study was designed to investigate the expression changes of osteopro-tegerin (OPG), tartrate-resistant acid phosphatase (TRAP) and vascular endothelial growth factor (VEGF) mRNA in collagen induced arthritis(CIA) rats. Methods After CIA was induced in Sprague-Dawley rats, the experimental animals were treated with PBS or rAAV-EGFP or rAAV-hOPG (100 μl/day) intra-articular injection for 10 days. Messenger RNAs (mRNAs) were obtained from CIA synovium 40days after first immun-ization. Reverse transcriptase-polymerase chain reactions (RT-PCR) were carried out to detect the mRNA encoding OPG, TRAP, VEGF and β-actin, which acted as inner control. The genes detected clearly by RT-PCR were quantified using real-time PCR. Results The expression of all genes was confirmed by specific single bands in RT-PCR. Real-time PCR showed that the expression levels of TRAP and VEGF were increased, whereas those of OPG mRNA were decreased in CIA group compared with normal controls. The intra-articular gene transduction markedly increased the gene copies of OPG by 128.21% (P<0.01). The expression change of OPG in synovium also caused the decrease of the expression levels of TRAP and VEGF by 58.79% (P<0.01)and 17.85% (P>0.05) respectively, however, the expression change of VEGF was not statistically significant. Conclusion OPG gene mediated by rAAV can be successfully tranfered to knee joint synovium in vivo. The results of this study suggest that gene transfer using rAAV-OPG may be a feasible and effective therapeutic approach to treat or prevent joint destruction in inflammatory arthritis.

Objective Using an in vivo adeno-associated virus(AAV)-mediated gene transfer technique,this study was designed to evaluate the protective effects of human osteoprotegerin(OPG)transgene against joint destruction in collagen induced arthritis(CIA)model.MethodsAfter CIA was established in the Sprague-Dawley rats,the experimental animals were treated with PBS or rAAV-EGFP or rAAV-hOPG (100μl/d)intra-articular injection 25 days after arthritis induction for 10 days.Paraffin-embedded joints were then analyzed histologically.The joint destruction was evaluated by Larsen Score.The protein expression of OPG,IL-1,MMP-3 was identified by enzyme-linked immunosorbent assay(ELISA).Results Suecessful trans-gene expression was confirmed by the detection of OPG by ELISA and positive fluorescence of the frozen joint section. Image analysis revealed that the expression of OPG significantly protected against joint destruction by 30% compared with the CIA group. Conclusion OPG gene transfer mediated by rAAV effectively protects against bone destruction induced by CIA model. Those data suggest that gene transferring using rAAV-OPG may be a feasible and effective therapeutic approach to treat or prevent joint destruction in inflammatory arthritis.

Objective To investigate the clinical characteristics of Macrophage activation syndrome (MAS) in patients with rheumatoid arthritis (RA), so as to reduce misdiagnosis. The objective of this paper was to improve the comprehensive and systematic understanding of rheumatoid arthritis complicated with macrophage activation syndrome. Methods The clinical data of one patient with macrophage activation syndrome secondary to RA were analyzed retrospectively, and the related literatures were reviewed. Results The patient was a 65 year old male with ahistory of RA for 14 years. The patient presented with symmetrical multi-joint pain aggravated with stiffness for 14 years and was admitted because of aggravation for 2 weeks. He failed many drugs for the treatment of rheumatoid arthritis was ineffective accompanied with intermittent leukocytopenia. After bone marrow aspiration and biopsy, the phenomenon of phagocytosis of macrophages was clearly diagnosed. He was treatment with high dose corticosteroid +CsA+ human immunoglobulin and his condition wasimproved. Literature was searched in PubMed, Wan Fang medical network database, RA+MAS. Finally, 12 related articles was yielded, and a total of 14 patients, including 8 males, 6 females. Four patients were adults and 10 were children. The shortest duration was 0.5 months, the longest was 24 months. Fever, skin rash, arthritis, enlargement of the liver or spleen, decreased of blood cells count, elevation of liver transaminase, increase of triglyceride, and a series of symptoms and laboratory parameters were observed in the course of the disease. Conclusion When rheumatoid arthritis patients show decreased blood leukocytes and can not be explained by other causes, the differential diagnosis should be carefully performed to rule out secondary macrophage activation syndrome. Always be awake of the risks and dangers of rheumatoid arthritis complicated with macrophage activation syndrome. Early diagnosis and timely are important to improve prognosis.

Objective To explore the molecular pathological mechanism of gout, and to explore the mechanism of how interleukin (IL)-37 influencing PDZK1 protein in gout through nuclear factor κB (NF-κB) pathway. Methods HK-2 cells were stimulated with inflammatory signal IL-37. The expression of PDZK1 and its subcellular localization were detected by real-time fluorescence quantitative polymerase chain reaction (real-time PCR) at different concentrations of IL-37 (defined as group A), PDTC+IL-37 (defined as group B), Wortmannin+IL-37 (defined as group C), respectively.The changes of PDZK1 protein expression in HK-2 cells were detected by adding inhibitor PDTC (NF-κB pathway inhibitor) or Wortmannin (PI3K pathway inhibitor) and inflammatory signal stimulating protein imprinting method. The comparative t test was used for statistical analysis between groups A, B and A and C. Results The average levels of PDZK1 mRNA were as follows:(group A: 28.71 ±0.35, 28.57 ±0.31, 28.78 ±0.28, 28.63 ±0.29, 28.62 ±0.19; group B: 28.71 ±0.31, 28.83 ±0.27, 28.58±0.26, 28.73±0.36, 28.68±0.35;group C:28.81±0.32, 28.91±0.29, 28.72±0.24, 28.59±0.18, 28.58±0.22). There was no significant change in PDZK1 mRNA level in group A and B. The same IL-37 was found in group A and B. The values of T were 5.73, 4.72, 4.69, 5.86 and 6.79, respectively. The P values were all greater than 0.05, and there was no significant difference between the two groups. The values of T were 6.78, 7.13, 7.47, 6.38 and 5.98 in the same IL-37 concentration groups of A and C, respectively, and the P values were all greater than 0.05, with no significant difference. The levels of PDZK1 protein in the three groups by Western blot analysis were as follows: (group A: 0.200±0.082, 0.412±0.032, 0.723±0.063, 1.202±0.021, 1.222±0.023;group B: 0.124±0.064, 0.303±0.081, 0.325±0.062, 0.223±0.071, 0.343±0.052; group C: 0.017±0.022, 0.204± 0.015, 0.187±0.053, 0.302±0.051, 0.404±0.051), The levels of PDZK1 protein in group A treated with different concentrations of IL-37 followed by the concentration of IL-37. The level of PDZK1 protein in group B increased with the increase of IL-37 concentration, but the level of PDZK1 protein in group B was lower than that in the group treated with IL-37 only, and decreased when the concentration of IL-37 was 40 ng/ml.The level of PDZK1 protein in group C increased with the increase of IL-37 concentration, but the level of PDZK1 protein in group C was lower than that in the group treated with IL-37 only, and the same concentration of IL-37 in group A and B. The values of T were 1.83, 1.37, 1.64, 1.57 ,1.49, with P values greater than 0.05. There was no significant difference between the two groups. The values of T were 1.28, 1.37, 1.26, 1.42, 1.39 in the same concentration of IL-37 in group A and C, with P values greater than 0.05, with no significant difference.The results of immunofluorescence analysis showed that the trend of the three groups was basically consistent with that of Western-Blot. Conclusion The pathogenesis of gout induced by IL-37 through PDZK1 protein may not occur at the transcriptional level, but may occur at the level of protein translation.