Objective To observe the expression and distribution of IL 10 in local vessel after balloon injury in rat and study its significance in the response of artery injury Methods RT PCR and Western Blot as well as immunohistochemical method were used to assay the changes of IL 10′s mRNA as well as expression and distribution of protein Results There is no expression of IL 10 in vascular wall of normal rat After Balloon injury, the level of IL 10 mRNA and product of IL 10 was up regulated, and immunohistochemiscal staining showed immunoreactive IL 10 mainly in smooth muscle cells Conclusion IL 10 is expressed in balloon injured aorta and may contribute to the modulation of the local inflammatory response

Objective To explore the role of receptor‐interacting protein kinase 3 (RIP3)‐mediated necroptosis in ulcerative colitis (UC ) . Methods The colonic mucosa tissues of eighty‐five patients diagnosed with UC were collected .Disease staging and pathological classification of UC patients were evaluated according to Mayo standard and Truelove standard ,respectively .The expressions of RIP3 and tumor necrosis factor (TNF)‐α were detected by immunohistochemistry .The cell inflammation model in HT‐29 cells was induced by interferon (IFN)‐γ.The expression of RIP3 mRNA and interleukin (IL)‐1βmRNA was determined by real time polymerase chain reaction (RT‐PCR) .Twenty healthy individuals were studied as controls .A single factor analysis of variance was performed in the comparison among groups ,and Pearson correlation analysis was used for correlation analysis .Results Among 85 patients with UC , 27 cases were mild ,30 cases were moderate and 28 cases were severe;for pathological classification ,26 were gradeⅠ ,31 were gradeⅡ and 28 were grade Ⅲ .The Mayo score was positively correlated with pathological grade (r=0 .997 , P<0 .05) .The expression of RIP3 and TNF‐α in colonic mucosa tissues of UC group increased along with the pathological classification grade .The expression of RIP3 of gradeⅠto Ⅲ was 1 .81 ± 0 .98 ,2 .77 ± 1 .26 and 4 .86 ± 2 .77 ,respectively ,and the differences were statistically significant (F=26 .10 ,P<0 .05) .The expression of TNF‐αof gradeⅠto Ⅲ was 1 .35 ± 0 .69 ,2 .61 ± 1 .41 and 4 .43 ± 2 .17 ,respectively ,and the differences were statistically significant (F=31 .80 ,P<0 .05) .The expression of IL‐1β mRNA of IFN‐γ group ,IFN‐γ+ z‐VAD group and IFN‐γ+z‐VAD+ TNF‐αgroup was 0 .68 ± 0 .27 ,1 .47 ± 0 .12 and 1 .86 ± 0 .16 ,respectively ,and the differences were statistically significant (F=38 .45 , P<0 .05) .The expression of RIP3 mRNA of IFN‐γgroup ,IFN‐γ+z‐VAD group and IFN‐γ+z‐VAD+TNF‐αgroup was 0 .46 ± 0 .13 ,1 .21 ± 0 .29 and 2 .06 ± 0 .20 ,respectively , and the differences were statistically significant (F= 55 .15 , P< 0 .05) .Conclusions TNF‐α/RIP3 signal pathway is up‐regulated in UC .RIP3‐mediated necroptosis promotes the death of colonic epithelial cells and exacerbates inflammatory response .RIP3‐mediated necroptosis may be an important mechanism in UC .

Objective:To establish the bacterial endotoxin test method and the abnormal toxicity test method for reduced glutathi -one for injection.Methods:According to the requirements and methods in Chinese Pharmacopoeia (2015 edition, part IV), the bacte-rial endotoxin test and the abnormal toxicity test for reduced glutathione were studied .Results:The limit of bacterial endotoxin for re-duced glutathione was 0.125 EU· mg-1 , and the limit of abnormal toxicity was 1.0 g· kg-1 .Conclusion: The bacterial endotoxin test method and the abnormal toxicity test method are feasible .The abnormal toxicity should be supplemented in the quality standard for reduced glutathione , and the bacterial endotoxin test can replace the pyrogen test .

Objective To investigate the effect of metformin on the proliferation and apoptosis in human pancreatic cancer cell line CFPAC-1,and to explore the potential mechanism.Methods Human pancreatic cancer CFPAC-1 cells were cultured in vitro,and were treated with metformin at different concentrations (1,2.5,5,10,20,40,60 mmol/L) for different durations (24 h,48 h and 72 h),and cells without treatment were considered as control group.Cell proliferation was evaluated by CCK-8,cell cycle was determined by flow cytometry,apoptosis was determined by Annexin V/PI double staining method,and Western blot was used to detect the protein expression of p-AMPK,FAS,cyclin D1,Bcl-xl,Bax,caspase-3 and survivin.Results Metformin could inhibit the proliferation of CFPAC-1 cells in a time and dose dependent manner.Forty-eight hours after 40 mmol/L metformin treatment,the proportion of CFPAC-1 cells in phase G0/G1 was significantly increased [(65.93 ± 0.27)％ vs (42.89± 1.02)％],and the proportion of CFPAC-1 cells in phase G2/M,S was significantly decreased [(22.01 ± 2.95) ％ vs (38.28 ± 4.93) ％,(13.58±0.43)％ vs (20.12 ± 3.38)％],and the difference between the two groups was statistically significant (P ＜0.05).The apoptosis rate was increased from (3.01 ± 0.49) ％ to (32.97 ± 3.19) ％,(P ＜ 0.05) ; and the expression of p-AMPK,Bax,and caspase-3 was increased,while the expression of FAS,cyclin D1,Bcl-xl,survivin were decreased.Conclusions Metformin can inhibit proliferation and promote apoptosis of CFPAC-1 cells mainly by activation of AMPK pathway,and down-regulation of FAS,cyclin D1,survivin and Bcl xl/Bax ratio,as well as up-regulation of caspase-3.

BACKGROUND:At present, spinal cord ischemia/reperfusion injury is considered as the main reason for secondary paralysis after spinal decompression, and to control the levels of stress-related proteins and excitatory amino acids plays an important role in the treatment of spinal cord ischemia/reperfusion injury. OBJECTIVE:To investigate the expression level of protein disulfide-isomerase A3 (PDIA3) after spinal cord ischemia/reperfusion injury in rabbits. METHODS:Thirty-six New Zealand white rabbits were enrolled, the models of spinal cord ischemia/reperfusion injury were established using Zivin's method, and were then randomized into six groups (n=6 per group). The rabbit abdominal aorta in control group was exposed without vascular occlusion and then the abdominal cavity was closed 30 minutes later. In experimental groups, the abdominal aorta was blocked for 30 minutes, followed by 0, 6, 12, 24 and 48 hours of reperfusion, and then the abdominal cavity was closed. The neurological function was evaluated with a modified Tarlov score. The L3-5lumbar vertebrae were removed, and PDIA3 was screened by two-dimensional fluorescence differential gel electrophoresis combined with mass spectrometry, and then its temporal and spatial changes in the spinal cord were detected by western blot assay and immunohistochemistry. RESULTS AND CONCLUSION:The function of hind limbs was improved in all the experimental groups after spinal cord ischemia/reperfusion injury, and the modified Tarlov scores reached the peak at 24 hours after schemia/reperfusion injury, and decreased slightly at 48 hours. The expression of PDIA3 in the control group showed clear imprinting, which was slightly strengthened at 0 hour, became more strengthened at 6-12 hours, significantly reduced to the minimum level at 24 hours, and returned to the level of 6-12 hours at 48 hours after ischemia/reperfusion. Immunohistochemical results showed that there was visible PDIA3 in the cytoplasm of neurons, and the expression level in the interneurons was significantly higher than that in the motor neurons. These results suggest that upregulated PDIA3 appears in the development and progression of spinal cord ischemia/reperfusion injury, indicating that PDIA3 is closely related to spinal cord ischemia/reperfusion injury, which can be used as a new diagnosis and treatment target.

Objective To compare the detection performance of the modified serum test (TRUST) method and the traditional method in syphilis screening.Methods A series of TRUST high titer syphilis serum was diluted,and the positive rate of each method was calculated by using the improved method and the traditional method.Comparison of two detection methods of C50,C5 ～ C95 interval,as well as the accuracy of the density curve,and the consistency of the two methods were compared,and diagnostic performance were compared.Results The improved method of C50 was less than the traditional method of C50,and the improved method of C5 ～ C95 range was narrower than the traditional method,compared with the traditional method.The improved method of the non precision density curve was steeper than the traditional method,and the two confidence interval of the consistency degree of the 95％ methods was 73.4％ to 95.8％.The diagnostic sensitivity (SEN),diagnostic specificity (SPE),positive predictive value (PPV),negative predictive value (NPV) and diagnostic efficiency(DF) of the improved method were 64.29 ％,99.1％,85.71％,97.05 ％ and 96.39 ％,respectively.The SEN,SPE,PPV,NPV and DF of the traditional methods were 3.75 ％,98.49 ％,75 ％,96.18 ％ and 95 ％,respectively.The improved method was superior to the traditional methods in the two aspects of SEN and PPV (x2 =8.25,10.03,all P＜0.05),with the statistically significant difference.The improved method was slightly higher than the traditional method in SPE,NPV and DF (x2 =2.39,3.45,4.03,all P＞0.05),with the no statistically significant difference.Conclusion The precision,diagnostic sensitivity and diagnostic specificity of the improved method was higher than that of the traditional method,and it can be applied to the detection of large batch samples with the aid of the full automatic enzyme immunoassay instrument.The improved method can be used to replace the traditional method for syphilis screening.

OBJECTIVE:To observe clinical efficacy and safety of Mailuoning injection combined with benazepril in the treat-ment of chronic glomerulonephritis. METHODS:130 patients with chronic glomerulonephritis were randomly divided into observa-tion group and control group,with 65 cases in each group. Both groups received rountine treatment as low salt and fat diet. Control group was additionally given Benazepril tablet 10 mg,qd;observation group was additionally given Mailuoning injection 20 ml added into 5% Sodium chloride injection 250 ml,ivgtt,qd,on the basis of control group,20 days of drug withdrawal every 10 days. Both group received 4 months of treatment. Clinical efficacy of 2 groups were observed as well as the levels of BUN,UCr, SCr and 24 h urine protein quantitative (UPE) before and after the treatment. The incidence of ADR was compared between 2 groups. RESULTS:Total effective rate of observation group was 83.1%,which was significantly higher than 67.7% of control group,with statistical significance (P<0.05). There was no statistical significance in BUN,UCr,SCr and 24 h UPE between 2 groups before treatment (P>0.05);those indexes of 2 groups decreased significantly after treatment,and the observation group were lower than the control group,with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups (P>0.05). CONCLUSIONS:Mailuoning injection combined with benazepril is effective for chronic glo-merulonephritis,and can effectively improve renal function with good safety.

AIM:To investigate the effect of batroxobin on thromboxane A 2(TXA 2)level in canine heart ischemia/reperfusion (I/R) injury and the cardioprotective mechanism of batroxobin against I/R injury. METHODS: Canine heart ischemia was induced by ligation of the left anterior descending coronary artery for 30 min followed by 90 min reperfusion.Batroxobin was intravenously administered before heart ischemia and 15 min before reperfusion.The level of plasma thromboxane (TXA 2) ,creatine phosphokinase (CK), lactic dehydrogenase (LDH ) and the concentration of myocardial TXA 2 were measured. The pathological changes in I/R myocardium were observed. RESULTS: In I/R group, TXA 2 levels of both plasma and myocardium increased significantly,and myocardium was injured obviously. Myocyte cells of central zone of I/R heart showed intracellular edema, swollen and damaged mitochondria, and fragmentation of cristae and concentrated nucleus. Plasma CK and LDH level was also elevated. Both ways of batroxobin administration reduced TXA 2 level apparently and plasma CK?LDH were also reduced significantly. LVEDP lowered while +dp/dt max elevated greatly,the mortality of I/R canine reduced obviously. CONCLUSION: Batroxobin decreased TXA 2 levels of both plasma and myocardium significantly, and could afford protection from I/R injury.

AIM: To observe the alteration of urotensin II (UII) receptors and contractile response to UII in rat aorta after balloon angioplasty injury. METHODS: The plasma membrane isolated from balloon injured aorta was used to study the binding of [ 125 I]-UII to the membrane and the contractile potency of UII on rat aorta was assayed. RESULTS: In contrast to the normal aorta, the contractile potency to UII enhanced in balloon injury artery and the calculated maximal number of specific binding sites (Bmax) was increased about 44% and 36% respectively in rat artery after balloon injury 3 and 21 days ( P

AIM: To observe the effect of adrenomedullin(ADM)on proliferation of vascular smooth muscle cells(VSMC) induced by urotensin Ⅱ(UⅡ). METHODS: DNA synthesis of cultured rat aortic VSMC was measured by -TdR incorporation. The activities of mitogen activated protein kinase(MAPK) were determined by isotope tagged with [?- 32 P]-ATP. RESULTS: UⅡ(10 -8 mol/L) significantly increased -TdR incorporation of VSMC and MAPK activities by 38%( P0.05 ), 32%( P0.05 ), 32%( P