OBJECTIVE: To provide proposal for government on dealing with the relationship between commercial and social property of drugs.METHODS: Multi-stakeholders subjects and their interrelation in the listing decision of drugs were investigated using game model.RESULTS & CONCLUSIONS: Innovation level of pharmaceutical industry (P),profitability of drugs operator (E),other factors affecting the proceeds of drug operators (K),additional gold cost of drugs (C1),additional cost of gold drugs sale (C2) were confirmed to be the key factors which affected the listing decision of drugs by pharmaceutical manufacturers.Governments are suggested to ensure the rational and fair use of drugs in public by policy supports,encouraging drug innovation and make-up on difference of drugs in medical institutions,optimizing benefit components of medical institutions,creating evaluation criteria for "listing value" and "necessity in clinic" of drugs.

Objective To investigate the association of single nucleotide polymorphisms(SNP)in p21and p27 genes with the risk of epithelial ovarian cancer(EOC).Methods Genotypes were analyzed by polymerase chain reaction-restrictive fragment length polymorphism(PCR-RFLP)method in 234 patients with EOC and 284 control women in China.Results (1)The frequencies of the p21 in healthy controls were 34.2%.49.6%and 16.2%,while the distribution of the C and T allele was 59.0%and 41.0%,respectively.The p21 C/C(28.2%),C/T(53.0%),T/T(18.8%)distribution in ovarian cancer patients was not significantly different from that in healthy controls(P＞0.05).There was no statistic difference in allele distribution between ovarian cancer patients and healthy controls(P＞0.05)either.The stratification analysis by tumor histological type did show that the genotype distribution in four types of ovarian cancer patients was significantly different from that in healthy controls(P=0.02).The C/C genotype was likely to reduce the risk of epithelial endometrial cancer.and the adjusted odds ratio was 0.56(95%CI:0.32-0.98).(2)The genotype frequencies of the p27 in healthy controls were 88.4%,10.9%and 0.7%.while the distribution of the V and G allele was 93.8%and 6.2%.respectively.The V/V(93.6%),V/G(5.1%)and G/G(1.3%)distribution in ovarian cancer patients was significantly different from that in healthy controls(P=0.04).There was no statistic difference in allele distributionbetween ovarian cancer patients and healthy controls(P＞0.05).Compared with the V/G and G/G genotypes,the V/V genotype increased the risk of EOC,the adjusted odds ratio was 1.92(95%CI:1.02-3.63).Conclusion The C/C genotype of p21 may reduce the risk of epithelial endometrial cancer,and the genotype of p27 V/V may be a potential risk factor for susceptibility to EOC.

BACKGROUND: Mesenchymal stem cells (MSCs) exist in umbilical cord blood (UCB), currently, there is not a method to in vitro separate, culture and amplificate human UCB-MSCs effectively. OBJECTIVE: To explore factors that influence yields of UCB-MSCs. METHODS: The relationship between the success rate of yielding UCB-MSCs and several factors, such as gestational ages (≥40 weeks, 37 weeks and ≤32 weeks), the number of mononuclear cells (MNCs) in UCB (≥2.5×109/L, <2.5×109/L), the inoculum density of MNCs (1×107, 1×109, 1×1011/L), the concentration of fetal bovine serum (FBS, 5%, 10%, 15%, 20%) in culture medium, and whether the culture flask being coated with FBS or not beforehand, as well as relationships among these factors were investigated. RESULTS AND CONCLUSION: The success rate of yielding UCB-MSCs was up to 58.3%. The success rate decreased as the gestational ages increasing (P < 0.01). The success rate could be enhanced to 76.9% when the MNCs count was more than 2.5×109/L, and there was significant difference when comparing to that of the group (36.4%) with MNCs count less than 2.5×109/L (x2=8.07, P=0.005). There was a negative correlation between the MNCs count and the gestational ages in the specimens with the same volume of UCB (r=-0.95, P < 0.01). In the group with the cell inoculum density of 1×1011/L, the growth and proliferation of primary and subculturing MSCs were better than that of the groups with the cell inoculum density lower than 1×1011/L. The adherence of MSCs in the group with the culture medium containing 5% FBS happened much later than other 3 groups, while the purity of MSCs in this group was much higher. When comparing the passage rate, there was no significant difference among the 4 groups with different concentration of FBS. In the group of culture flask being coated with FBS beforehand, the purity and proliferation ability of MSCs was higher than that in the groups with culture flask not being coated. It is suggested that culture of UCB-MSCs was influenced by several factors. The success rate could be increased by choosing the fetus with relative lower gestational ages, collecting enough volume of UCB, inoculating cells with a higher density, choosing the medium with lower concentration of FBS, and coating the culture flask with FBS beforehand.

Objective To investigate whether fasting obestatin level is different in patients with impaired glucose tolerance or type 2 diabetes, and to explore the association between obestatin and lipid metabolism. Methods Eighty-four subjects without known diabetes were divided into three groups: normal glucose tolerance(NGT), impaired glucose tolerance (IGT) and type 2 diabetes (DM) Plasma obestatin levels were measured with a radioimmunoassay. The relationship between fasting obestatin levels and metabolic parameters was also analyzed. Results Fasting obestatin levels were lower in DM group [(2.82±0.78)ng/ml] and IGT group [(3.25±0.29)ng/ml] than in NGT group[(3.55±0.57) ng/ml, P＜0.01]. Triglycerides and low density lipoprotein cholesterol levels gradually increased among the three groups (P＜0.05). Multiple linear regression analysis revealed fasting obestatin level was independently associated with waist-to-hip ratio, triglyeride and low density lipoprotein cholesterol. The regression equation was obestatin=6.953-3.412×W/H-0.175×TG-0.123×LDL-C. Conclusions The decreased obestatin may be associated with IGR and T2DM, and obestatin level may be associated with lipid metabolism.

Objective To study the effects on berberine hydrochloride on the mitochondria membrane potential and free intracellular calcium of HaCaT cells, and elucidate the mechanism of action of berberine on keratinocytes. Methods Rhodamine-123 fluorescence (very sensitive to mitochondria membrane potential) and Fluo-3/AM fluorescence (suitabe to detect free intracellular calcium in single HaCaT cell) were measured by laser scanning confocal technique. Results Fluo-3/AM fluorescence intensity of HaCaT cells was persistently increased after treating with berberine at concentrations of 5 ? 10-5M, 2.5 ? 10-5M and 1.25 ? 10-5M, and significant differences were observed as compared with the PBS control. The intensity of rhodamine-123 fluorescence in HaCaT cells was decreased immediately when exposed to berberine, with significant difference from that of the PBS control. Conclusions It is suggested that berberine could increase free intracellular calcium and decrease mitochondria membrane potential of HaCaT cells, induce overload of intracellular calcium, influence energy metabolism, and then inhibit the proliferation of keratinocytes.

Objective To investigate the protective effect of ischemia preconditioning(IPC)on apoptosis in-duced by renal ischemia - reperfusion(IR)and relations to the changing expressions of Bcl - 2,Bax in rat kidney. Methods Ischemia models were induced by clipping bilateral renal pedicles for 30 min by using the artery clamp;IPC group was induced by clipping bilateral renal pedicles for 15 min,4 days later IR was performed again by clipping bila-teral renal pedicle for 30 min. Rats were randomly divided into 5 groups with 5 animals in each group:control group(C group),sham - operation group(S group),IR group,IPC group(IPC ﹢ IR group),sham IPC group(S ﹢ IR group),all groups were randomly divided into 9 sub groups(0 h,3 h,6 h,12 h,24 h,48 h,3 d,5 d,7 d)except C group according to the time points after reperfusion. Occurrence of apoptosis was detected by terminal deoxynuleotidyl transferase media-ted dUTP nick end and labeling(TUNEL)method;the mRNA expression and protein levels of Bax and Bcl - 2 were de-tected by reverse transcriptase - polymerase chain reaction and quantitave immunohistochemisty. Results (1)Com-pared with S group and S ﹢ IR group,serum creatinine,blood urea nitrogen,kidney pathological damage scores in IR group gradually increased after IR,and peak point was 24 h after reperfusion;among all the subgroups there was a sig-nificant difference(all P ﹤ 0. 01). The expression of Bax,Bcl - 2 mRNA raised sharply in IR group after reperfusion, peaking at 6 h,24 h of reperfusion respectively,2. 66 ± 0. 12,2. 70 ± 0. 10,and among all the subgroups there was a sig-nificant difference(all P ﹤ 0. 01);the expression of Bax,Bcl - 2 protein had significant difference(all P ﹤ 0. 05). TUNEL immunofluorescence staining showed C group and S group had no obvious apoptosis cells in renal tubular epi-thelium;epithelial cell apoptosis after IR gradually increased in IR group,peaking at 24 h of reperfusion[(25. 07 ± 2. 29)% ].(2)Compared with IR group and S ﹢ IR group,pathological injury was significantly decreased in IPC ﹢ IR group;the expression of Bax,Bcl - 2 mRNA and protein,apoptosis cells were significantly decreased in IPC ﹢ IR group (all P ﹤ 0. 05). Conclusions Bax,Bcl - 2 are closely associated with kidney injury induced by IR. IPC may regulate acute kidney injuries by regulating Bax/ Bcl - 2.

Objective To investigate the expression and significance of microRNA - 21(miR - 21)in acute kidney injury mice model at the different time points following ischemic/ reperfusion. Methods C57BL/ 6J mice were divided into 3 major groups:the control group(C group),sham operation group(S group)and ischemia - reperfusion group(IR group). Later 2 groups were divided into 9 sub - groups respectively according to the time following reperfu-sion. Automatic biochemical analyzer detected serum creatinine(Scr),blood urea nitrogen(BUN)level. HE staining detected renal pathological damage. Renal tubulointerstitial pathological score accessed pathological damage. Real time - PCR tested the expression of miR - 21 and mitogen - activated protein kinase kinase 3(MKK3)mRNA in renal respectively. Immunohistochemistry staining tested expression of MKK3. Results IR group's Scr,BUN levels gradually increased following reperfusion,24 h reached its peak,then gradually declined. The Scr,BUN level had statistically sig-nificant difference between IR group and S group at the same time subgroup from 3 h to 168 h following reperfusion(all P ﹤ 0. 01). The change of kidney damage and pathological changes of interstitial and tubular injury score consensus with renal function. miR - 21 increased gradually in renal ischemia after reperfusion,24 h peaked and then stabilized at this high level. miR - 21 was positively correlated with pathological tubulointerstitial injury from 0 h to 168 h after reperfu-sion(r = 0. 969,P ﹤ 0. 05). IR group's MKK3 mRNA and protein expression rose sharply following ischemia/ reperfu-sion,24 h peaked,and then gradually decreased. From 3 h to 168 h,the expression of MKK3 mRNA and proteins had significant difference at each same time points subgroups between IR group and S group(all P ﹤ 0. 01). Conclusions miR - 21 increases gradually in renal ischemia after reperfusion,24 h peaked and then stabilized at this high level. miR - 21 is positively correlated with pathological tubulointerstitial injury,which may be associated with the negative regulated relationship between miR - 21 and MKK3.

Objective To investigate the relationship between anti-Helicobacter pylori (Hp) antibodies and development of chronic urticaria (CU).Methods Fifty CU patients with positive 13C-urea breath test and anti-Hp antibodies,as well as 50 healthy human controls were recruited in this study.Serum samples were collected from all the subjects.The samples from the patients were subjected to tests for anti-high affinity IgE receptor (anti-FcεRI) and-IgE antibodies.Human mast cells (HMCs) were classified into several parts to be incubated with the sera of patients with CU,the sera of healthy controls with anti-IgE and-FcεRI antibodies respectively for 20 minutes.Those incubated with the sera of healthy controls without these antibodies served as the control.Subsequently,the levels of histamine released by HMCs were measured by enzyme-linked immunosorbent assay (ELISA).Results The sera of CU patients showed a stronger ability to activate HMCs to release histamine than those of healthy controls ((3.13 ± 0.93) μg/L vs (2.92 ± 0.75) μg/L,t =2.39,P ＜ 0.05).Anti-FcεRI antibodies were detected in 4 patients,and antiIgE antibodies in 3 patients.A significant increase was observed in the levels of histamine released by HMCs incubated with anti-FcεRI antibody-positive and anti-IgE antibody-positive patient-derived sera (t =4.82,6.34,respectively,both P ＜ 0.01),but not in those incubated with patient-derived sera only positive for anti-Hp antibodies (t =1.74,P ＞ 0.05) compared with those incubated with healthy control-derived sera.In comparison with the antibody-free healthy control-derived sera,those with anti-Hp IgG antibodies showed no significant effect on the release of histamines by HMCs (t =1.95,P ＞ 0.05),whereas those with anti-FcεRI antibodies and anti-IgE antibodies exhibited an obvious promoting effect (t =3.72,3.02,respectively,both P ＜ 0.01).Conclusions The anti-Hp antibodies appears to have no role in the pathogenesis of CU,but the presence of anti-FcεRI and anti-IgE antibodies may contribute to the initiation of CU in patients with Hp infection.

OBJECTIVE To analyze the resistance of Enterobacter cloacae AmpC ?-lactamase with/without being induced by imipenem(IMP),cefepime(FEP),ceftriaxone(CRO) and aztreonam(ATM).The ampD genes of mutant strains were sequenced. METHODS Five wild type strains and 5 hyper-inducible type strains of E.cloacae were selected and induced by the tested antibiotics in vitro.At the same time,antibiotic susceptibility tests,3-D test,isoelectric focusing(IEF) and inhibit experiment were detected to identify hyper-producing AmpC ?-lactamase.ampD Gene sequencing was preformed in part of mutant strains. RESULTS There wasn′t obvious alteration in MIC with IMP,FEP,CRO and ATM for wild type strains whether they were induced or not.But,for the hyper-inducible type,there was apparent increasing in MIC after antibiotics inducing,especially CRO and ATM,up to 10.7-128 times.The DNA sequences analysis in the mutation strains showed there existed the replacement of the single base in multiple sites,and a few sites of amino acid were altered. CONCLUSIONS Mutant sites in ampD gene sequences are identical even though antibiotics as inducer are different.

Objective To estimate the correlations between chronic idiopathic urticaria (CIU) development and interrelated autoantibodies,including anti-high affinity immunoglobulin E receptor (anti-FcεRI) antibody,anti-immunoglobulin E (anti-IgE) antibody,anti-Helicobacter pylori (HP) antibody and antithyroglobulin antibody (TGAb).Methods This study included 100 patients with CIU,100 patients with acute urticaria (AU) and 100 healthy controls.Autologous serum skin test (ASST) was performed and allergens were detected by fluorescence-based enzyme linked immunosorbent assay (ELISA) in each subject.Serum levels of total IgE,anti-FcεRI antibody,anti-IgE antibody,anti-HP antibody and TGAb were measured.Chi-square test,analysis of variance,and Wilcoxon rank sum test were conducted for statistical analysis.Results The positivity rate of ASST was 53％,12％ and 0 respectively in patients with CIU,patients with AU and healthy controls,respectively.Food or inhalant allergens were detected in 86％ of the patients with AU,but not detected in any of the patients with CIU or healthy controls.Patients with CIU showed significantly higher levels of anti-FcεRI antibody and anti-IgE antibody compared with patients with AU and healthy controls (all P ＜ 0.05).The serum IgE level in healthy controls was statistically lower than that in patients with AU (T =226.00,P ＜ 0.05),but higher than that in patients with CIU (T =190.00,P ＜ 0.05).ASST-positive patients with CIU had a higher level of serum anti-FcεRI antibody (T =101.73,P ＜ 0.05),but a similar level of serum anti-IgE antibody compared with ASST-negative patients with CIU (T =312.04,P ＞ 0.05).No significant differences were observed in the positivity rate of anti-HP antibody (29％,19％ and 23％,P ＞ 0.05) or TGAb (18％,15％ and 11％,P ＞ 0.05) between the patients with CIU,patients with AU and healthy controls.Both anti-HP antibody-positive patients and TGAb-positive patients with CIU showed a significantly higher positivity rate of anti-FcεRI antibody (all P ＜ 0.01),but a similar positivity rate of anti-IgE antibody compared with the patients with AU and healthy controls (all P ＞ 0.05).Conclusions Anti-FcεRI antibody and anti-IgE antibody are present in patients with CIU,and may play a certain role in the pathogenesis of CIU.