The present study aimed to establish an LC-MS/MS method for the quantification of tiamulin in minipig plasma and to further conduct a pharmacokinetic comparison of different formulations. The plasma samples were extracted with acetonitrile (meloxicam as internal standard), separated on a C18 column, and quantified by multiple reaction monitoring mode (ESI+). Sanyuan minipigs were used as experimental animals. Plasma samples were collected after intravenous injection (10 mg/kg) and intragastric administration (20 mg/kg). The method showed good linearity, with intra- and inter-batch RSD of 1.00%–8.13% and RE within ±15%. The extraction recovery, matrix effect and stability of the analytical methods met the relevant requirements. Tiamulin fumarate active pharmaceutical ingredient was intravenously administered, with c0 of about (4383.73±2676.78) ng/mL, AUC0-t of about (4803.50±965.68) h·ng/mL, t1/2 of about (4.66±1.68) h, and CL of about (2.14±0.46) L/(kg·h). Three tiamulin formulations were intragastrically administered, with cmax of (552.00±328.55), (545.00±136.97) and (590.60±237.02) ng/mL, tmax of (1.47±0.68), (0.69±0.75) and (0.72±0.72) h, and F of 24.85%, 15.28% and 16.97%, respectively. The validated method meets the requirements for biological sample analysis and is applicable for the pharmacokinetic evaluation of tiamulin formulations in minipigs.

Objective To investigate the effects of transplantation with different amount of bone marrow mesenchymal stem cells(BMSCs)on osteoporosis in ovariectomized rats.Methods The rat model of osteoporosis was prepared by ovariectom(OVX)and verified after 3 months.A total of 60 female Sprague-Dawley rats were randomly divided into 6 groups(n=10 each).The rats with shamed operation served as a sham-injured control group(sham control group).The 5 ovariectom (OVX) groups with osteoporosis were study groups as follows:(1)the negative therapy group(simple OVX group);(2)the positive therapy group(OVX+ estrogen,E2 group) by intramuscular injection;others were treated with transplantation of BMSCs by tail vein injection in low dose(LS group),middle dose (MS group)and high dose(HS group).At 8,12 and 16 weeks after intervention,body mineral density (BMD)were detected by dual-energy x-ray absorptiometry scans.After 16 weeks of intervention,rat shinbone was obtained and stained by hematoxylin-eosin(HE) staining.Results Compared with the sham control group,simple OVX group showed a reduced total body BMD and the decreased proportion of trabecular bone to bone marrow cavity area (P ＜0.05).The total body BMD and the proportion of trabecular bone to bone marrow cavity area were higher in each BMSCs transplantation groups than in simple OVX group at 8,12,16 weeks after intervention(P ＜0.05),which showed a increased trend in the total body BMD and the proportion with the increased dose of transplantation BMSCs(P＜0.05).Rats in the HS group had highest BMD and best proportion of trabecular bone to bone marrow cavity area among three doses of transplantation BMSCs.Conclusions BMSCs transplantation can significantly improve osteoporosis of ovariectomized rats with an increased total body BMD and higher proportion of trabecular bone to bone marrow cavity area,and better and longer outcomes can be achieved.

Objective To investigate the improvement of symptoms of the patients after treatment in patients with Rome Ⅳ or non-Rome Ⅳ irritable bowel syndrome (IBS),and to explore the influence of IBS diagnosed by different criteria on the patients.Methods From June 2nd to 8th in 2016,at Outpatients Department of Gastroenterology,Union Hospital Affiliated to Tongji Medical College,Huazhong Uiversity of Science and Technology in Wuhan,1 500 outpatients aged over 18 years old and with intestinal symptom were selected for questionnaire.After treatment for six months,IBS patients,non-IBS patients,patients with Rome Ⅳ IBS and patients with non-Rome Ⅳ IBS were followed up by phone calls.After treatment,the improvement of symptoms of the patients was evaluated by irritable bowel syndrome symptom severity scale (IBS-SSS).The degree of influence of IBS diagnosed with different criteria on patients was evaluated by the patient's daily work whether to choose colonoscopy examination,whether to choose medication,and the efficacy of medicine.Student's t test,Mann-Whitney U test and chi-square test were performed for statistical analysis.Results A total of 352 patients with intestinal symptoms were followed-up,including 175 patients with IBS (84 patients with Rome Ⅳ IBS and 91 patients with non-Rome Ⅳ IBS) and 177 non-IBS patients,and 142 patients responded.There were no statistically significant differences in response rate between non-IBS patients and IBS patients (37.3％,66/177 vs.43.4％,76/175),and between patients with Rome Ⅳ IBS and patients with non-Rome Ⅳ IBS (40.5％,34/84 vs.46.2％,42/91) (x2 =1.379 and 0.573,P =0.240 and 0.449).Compared with the non-IBS patients,the degree of satisfaction of medicine was lower in IBS patients (71.4％,30/42 vs.47.5％,19/40).Compared with non-Rome Ⅳ IBS patients,Rome type Ⅳ IBS patients were more likely to receive colonoscopy (35.7％,15/42 vs.58.8％,20/34),and the differences were statistically significant (x2 =4.878 and 4.039,P =0.027 and 0.044).After six months of treatment,symptoms improved in both Rome Ⅳ IBS patients and non-Rome Ⅳ IBS patients (both P ＜ 0.05),however,the symptoms improved more significantly in Rome Ⅳ IBS patients and the total score of IBS-SSS was lower than that of non-Rome Ⅳ IBS patients (-130,-185 to 60 vs.-70,-100 to 28),and the difference was statistically significant (Z =-3.065,P =0.002).The difference was mainly showed the symptom of abdominal pain,and the IBS-SSS abdominal pain score of Rome Ⅳ IBS patients was lower than that of non-Rome Ⅳ IBS patients (-80,-100 to-40 vs.0,-40 to 0),and the difference was statistically significant (Z =-4.631,P ＜ 0.01).Conclusions IBS symptoms influence a lot on the satisfaction degree of treatment in outpatients.Even with similar good therapeutic effects,the Rome Ⅳ IBS symptoms have a more severe impact on patients than non-Rome Ⅳ IBS symptoms.

Objective To explore the impact factors of postpartum depression.Methods A total of 257 patients from October 201 4 to April 201 5 were investigated with postpartum depression screening scale and self-designed questionnaire.The results were analyzed by statistical software.Results The total rate of postpartum depression was 1 4.4%.The education level,family relationship,newborn sex,perinatal knowledge learning,spousal relationship and delivery mode were independent factors for postpartum depression,which were statistically different between depression group and normal group (P <0.05 ).Conclusions Poor family relationship especially poor spousal relationship,low education level,lack of perinatal knowledge,female newborn and caesarean section increase the risk of postpartum depression.

Objective To investigate the distribution of alpha2-HS glycoprotein (AHSG) gene polymorphisms and the relationships of AHSG gene polymorphisms with atherosclerosis as well as serum bone related biochemical mark-era. Methods Blood samples of 344 hospitalized female patients, aged 20 ～ 80 years, were sampled for serum bone alkaline phosphatase, cross-linked N-telopeptide of collagen type Ⅰ, cross-linked C-telopeptide of collagen type Ⅰ , osteoprotegrin and leptin were determined by ELISA. Serum TC,TG and calcium content were detected. Poly-morphism of AHSG gene was detected by polymerase chain reaction fragment length polymorphisms (PCR-RFLP) of restriction enzyme Sac Ⅰ. BMD (Norland XR-36) of the anteroposterior spine (AP), supine lateral spine (Lat) and femoral neck (FN) were measured. Morphological changes in the aorta and bone of type GG patient were detected by pathological microscopy. IMT were measured by color doppler ultrasound equipment(SEQUOIAS12). Results (1) The genotype frequency of CC, CG, and GG were 59.7%, 25.1% and 15.2% respectively in all elderly female patients. There were significant difference in blood lipids, Ca~(2+) and serum bone relative biochemical markers to different AHSG genotypes. (2)There were significant differences in the BMD of the AP, Lat, FN and IMT and the serum biochemical markers among the CC, CG and GG genotypes. (3)GG-female patients bone tissue pathology section verify the AHSG polymorphism genetic mutation and atherosclerosis, osteoporosis development of the relationship. Conclusion There was close relationship among AHSG polymorphism variation and the incidence of arteriosclerosis and osteoporosis in elderly female.

Objective To investigate the distribution of alpha2-HS glycoprotein(AHSG) gene polymorphisms and the relationships of AHSG gene polymorphisms with atherosclerosis as well as serum bone related biochemical markers.Methods Blood samples of 344 hospitalized female patients,aged 20～80 years,were sampled for serum bone alkaline phosphatase,cross-linked N-telopeptide of collagen typeⅠ,cross-linked C-telopeptide of collagen type Ⅰ,osteoprotegrin and leptin were determined by ELISA.Serum TC,TG and calcium content were detected.Polymorphism of AHSG gene was detected by polymerase chain reaction fragment length polymorphisms(PCR-RFLP) of restriction enzyme Sac Ⅰ.BMD(Norland XR-36) of the anteroposterior spine(AP),supine lateral spine(Lat) and femoral neck(FN) were measured.Morphological changes in the aorta and bone of type GG patient were detected by pathological microscopy.IMT were measured by color doppler ultrasound equipment(SEQUOIA512).Results(1) The genotype frequency of CC,CG,and GG were 59.7%,25.1% and 15.2% respectively in all elderly female patients.There were significant difference in blood lipids,Ca2+ and serum bone relative biochemical markers to different AHSG genotypes.(2)There were significant differences in the BMD of the AP,Lat,FN and IMT and the serum biochemical markers among the CC,CG and GG genotypes.(3)GG-female patients bone tissue pathology section verify the AHSG polymorphism genetic mutation and atherosclerosis,osteoporosis development of the relationship.Conclusion There was close relationship among AHSG polymorphism variation and the incidence of arteriosclerosis and osteoporosis in elderly female.

Objective To evaluate the prevalence and risk factors of peripheral arterial obstructive disease(PAD) in type 2 diabetes mellitus(T2DM) patients. Methods A total of 401 patients with T2DM(201 men and 200 women),mean aged was(63.4?8.8)years.PAD was diagnosed by an ankle-brachial index(ABI)≤0.9 on either leg.Results Overall prevalence of PAD was 19.4%,without significant sexual difference.Univariate analyses disclosed age,hypertension,systolic blood pressure(SBP),low dense lipoprotein cholesterol(LDL-C) and T2DM duration as significant risk factors.In multiple Logistic regression,age,SBP,LDL-C and T2DM duration were independent risk factors with respective odds ratios(95% confidence intervals,CI) of 1.94(1.20-2.60),1.12(0.88-1.43),1.32(0.97-1.63) and 1.27(0.96-1.73) respectively.Conclusions Prevalence of PAD in T2DM patients is(19.4)% and the major risk factors were older age,higher SBP,higher LDL-C and T2DM duration.This implies that it is very important to prevent the incidence of PAD for better glucose,lipid and blood pressure control early.

Objective To investigate the changes between type 2 diabetes mellitus(T2DM) with peripheral vessel disease and osteoporosis. Methods A total of 100 cases T2DM patients were divided into non-peripheral vessel disease (A group) and peripheral vessel disease (B group). B-mode ultrasonography to measure the intima-media thickness(IMT) of carotid artery,dual energy X-ray absorptiometry (DEXA) to measured bone mineral density(BMD) and VS-1000 arteriosclerosis diagnometer to measure brachi-ankle pulse wave velocity (baPWV) were used in 100 T2DM patients. The clinical data and BMD were compared between two groups. Results Compared with normal group,the BMD in T2DM patients significantly decreased.Compared with T2DM patients without peripheral vessel disease,the BMD decreased in T2DM patients with peripheral vessel disease. Conclusions Atherosclerosis in T2DM with peripheral vessel disease is probably related to osteoporosis, and the relationship between them is worth further investigation.

Objectives　To characterize the activation of platelet integrin αⅡbβ3 induced by two anti-human platelet tetraspanin monoclonal antibodies (mAbs), HI117 and SJ9A4, and investigate their potential mechanism of action. Methods　Using 125　　I-labeled human fibrinogen (Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured. Results　HI117 and SJ9A4 (10?μg/ml and 20?μg/ml) induced specific Fg binding to human platelets, suggesting that the two mAbs evoked activation of platelet integrin αⅡbβ3. Further study indicated that HI117 and SJ9A4 induced integrin αⅡbβ3 activation independent of platelet Fc-receptors, and that HI117 and SJ9A4-induced integrin αⅡbβ3 activation was inhibited by pretreatment of platelets with sphingosine, aspirin, apyrase, and/or PGI2. Conclusions　Anti-platelet tetraspanin (CD9) mAbs, HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 activation independent of Fc-receptors. Three signaling pathways, namely thromboxane, secreted ADP, and cAMP pathways, may be involved in the process, with protein kinase C activation presumably being the common step of the three pathways.

Objective. This study characterized the activation of platelet integrin aⅡbβ3 induced by two anti-human platelet te-traspanin monoclonal antibodies(mAbs),HI117 and SJ9A4. Methods. Using 125I-labeled human fibdnogen(Fg), specific Fg binding to human platelets induced by HI117 and SJ9A4 was measured as indication of activation of platelet integrin αⅡbβ3 by the two mAbs. Results. H1117 and SJ9A4( 10μg/ml and 20μg/ml) induced evident specific Fg binding to human platelets, sug-gesting that the two mAbs evoked activation of platelet integrin αⅡbβ3. Further study indicated that HI117 and SJ9A4 induced integrin t Ⅱ 1β3 activation independent of platelet Fc-receptors, and that HI117 and SJ9A4-induced integrin αⅡbβ3 activation was inhibited by sphingosing, aspirin, apyrase, and/or PGI2. Conclusion. The anti-platelet tetraspanin(CD9)mAbe,HI117 and SJ9A4, can induce platelet integrin αⅡbβ3 act-vation independent of Fc-receptors. Three signaling pathways,i.e.thromboxane,secreted ADP, and cAMP pathways may be involved in the process, with protein kimse C activation presumably being the comtmon step of the three pathways.