This paper adopted a series of related analysis methods to comprehensively analyze post-marketing clinical safety data of Shenmai injection from 4,220 cases of SRS and 32,358 cases of multicenter, prospective, registered hospital centralized monitoring in large data background, calculated ADR incidence rate was 0.93 per 1,000, main symptoms of ADR includes chest pain, chills, skin itching, palpitations, fever, nausea, dizziness, vomiting, flushing, numbness, allergic reaction, cyanosis, rash, low back pain, and "breath", "anaphylactoid reaction" and "flush" were the safety warning signals of Shenmai injection. Primary disease for chronic pulmonary heart disease, thyroid disease, and combined with cerebral vascular disease, prior to the injection and continuous use of alprostadil, cyclic adenosine monophosphate, combined with quinolones, penicillins were suspicious influence factors of ADR of Shenmai injection, these promot the clinical safety.
Drug Combinations ; Drugs, Chinese Herbal ; adverse effects ; Humans ; Injections ; Product Surveillance, Postmarketing ; Prospective Studies

UNLABELLEDTo reduce the huge labor-cost in the screening in traditional monoclonal antibody generation, We established a new system for monoclonal antibody generation integrating with protein array. BALB/c mice were immunized by eight recombinant proteins respectively, and the positive hybridoma cells were obtained by cell fusion and ELISA screening. All the eight kinds of positive hybridoma cells were mixed, cloned, screened by protein array, and definite dilution cloned.

RESULTS175 single cell clones were obtained by complex cloning, and 119 of those were positive clones. Then 8 positive cell lines were generated by the following 2 rounds definite dilution cloning. By comparing with the traditional method, we got 8 monoclonal antibodies using the combined protein array screening and multiplex cloning method in 1 cycle, and fewer amounts of antigens were used. As a result, the combined protein array and multiplex cloning method could be used as an economical, rapid and simple tool applying in high throughput monoclonal antibody generation.

Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; isolation & purification ; Female ; Humans ; Hybridomas ; metabolism ; Mice ; Mice, Inbred BALB C ; Protein Array Analysis

OBJECTIVETo observe the homing and differentiation of marrow-derived mesenchymal stem cells (MSC) transplanted intravenously in smoke inhalation injured rabbits.

METHODSThirty-two New Zealand big ear rabbits were divided into normal control group (NC), inhalation injury group (II), normal control + MSC treatment group (NM), and MSC treatment group (MT) according to the random number table, with 8 rabbits in each group. Rabbits in NC group were injected with 10 mL phosphate buffered saline (PBS) via ear marginal vein. Rabbits in NM group were injected with 10 mL PBS containing the third generation MSC labeled by BrdU (1 × 10(7) per 10 mL PBS) via ear marginal vein. Severe smoke inhalation injury model was reproduced in the other two groups, among them rabbits in II group were treated as rabbits in NC group, rabbits in MT group treated as rabbits in NM group. On the 7th and 28th day post treatment (PTD), lung tissue and trachea tissue were harvested from four groups for observation on injury with HE staining. Homing of MSC in injured tissue was observed with immunohistochemistry staining. The differentiation of MSC into functional cells was observed with immunohistochemical double staining of combining nuclear marker BrdU with lung (trachea) membrane-specific marker aquaporin-5 (AQP-5), alkaline phosphatase (AKP), CD34, and cytokeratin respectively.

RESULTS(1) MSC homing in lung and trachea tissue was observed in MT group on PTD 7, which was not observed in NM group. (2) AQP-5, AKP, and CD34 positive MSC were observed in lung tissue in MT group on PTD 28, while cytokeratin positive MSC was not observed in trachea tissue. No positively marked MSC was observed in NM group. (3) Injury in lung and trachea was less severe in MT group than in II group; and the proliferation of fibroblasts was less in MT group.

CONCLUSIONSIntravenous injection of MSC to rabbits with smoke inhalation injury can migrate to lung and trachea tissue at obviously inflammatory site, and differentiate into alveolar epithelial cells typeI and II, and pulmonary vascular endothelial cells, which may participate in the process of tissue repair in smoke inhalation injury.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Endothelial Cells ; cytology ; Epithelial Cells ; cytology ; Lung ; cytology ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; cytology ; Pulmonary Alveoli ; cytology ; Rabbits ; Smoke Inhalation Injury ; pathology ; Trachea ; cytology

Taking the genome DNA of Infectious Bovine Rhinotracheitis Virus (IBRV) as the template, the gG gene was amplified with PCR and cloned into the T cloning vector pMD18-T. After being identified by restriction digestion and DNA sequencing, the insert was subcloned into the expression vector pGEX-KG. Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that this gene was expressed as both soluble form and inclusion body by the transformed E. coli BL21 strain (DE3). The fusion protein was purified and used as the coating antigen to develop the indirect Enzyme-Linked Immunosorbent Assay (ELISA). Comparison between this gG-ELISA and commercial IBRV gB-ELISA Kit (IDEXX) was made in the detection of 380 cow serum samples. The results demonstrated an agreement of 92%. By using this novel gG-ELISA, 1248 cow serum samples were tested and the average positive rate of IBRV antibodies for imported cows is 21.7%, while the positive rate ranged greatly from 0.0%-41.5% for Hubei local Chinese Black and White Dairy Cows.
Animals ; Antibodies, Viral ; blood ; immunology ; Antigens, Viral ; genetics ; immunology ; Cattle ; Cloning, Molecular ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; genetics ; metabolism ; Female ; Herpesvirus 1, Bovine ; genetics ; immunology ; Male ; Recombinant Proteins ; biosynthesis ; genetics ; immunology ; Sensitivity and Specificity ; Viral Proteins ; biosynthesis ; genetics ; immunology

OBJECTIVETo study the effect of BAM bone grafting combined with inactivated autologous porous bone flap in repairing skull defect in rats.

METHODSSeventy-two Wistar rats with skull defect were randomly divided into control group, inactivated autologous bone flap group (AB group), BAM bone-induced artificial bone material group (BAM group), and inactivated autologous bone flap with BAM bone-induced artificial bone group (BAM+AB group). The bone healing was evaluated with micro-CT and the new bone formation was assessed with histological staining at 1, 2, and 3 months after modeling.

RESULTSInactivated porous bone flap combined with BAM bone-induced artificial bone effectively induced vascular and fibrous tissue regeneration and osteogenesis in the cranial defects. With the inactivated porous bone flap as the scaffold, BAM bone-induced artificial bone obviously promoted the restoration of the skull appearance in the rats with cranial defects.

CONCLUSIONInactivated autologous bone flap group and BAM bone-induced artificial bone material can promote skull healing and restoration of the original skull appearance, and can be used for reconstruction of the local anatomy of the skull surface.

Objective To investigate the employers' and employees' satisfaction of Zhejiang Province on occupation health examination and diagnosis of occupational diseases, and to guide and to standardize the occupation health examination and occupational disease diagnosis. Methods A random sample of 953 employers, 1791 workers with health examination and 135 workers with diagnosis of occupational diseases were selected in the survey, and the questionnaire about the Satisfuction on occupation health examination and occupation disease diagnosis were used in this survey. Results A total of 2879 questionnaires were sent out, in which 2841 valid questionnaires were returned, and the effective recovery rate was 98.68%. The recognition rates on comfortable environment, clear instructions process, workflow notification, and attention notification were all above 98%. The satisfaction rates for all items were above 86%, and the total satisfaction rate was 89.27% . The total satisfaction rates of workers with health examination, workers with diagnosis of occupational diseases and employers were 89.28%, 82.03%, and 90.22%, respectively. The recognition rates on clear instructions process and attention notification, and the satisfaction rates on service attitude, result information and overall satisfaction were significantly different between different types of respondents (P<0.05) . The results of pair wise comparison showed that the satisfaction rates of workers with diseases diagnosis on service attitude, results information and overall satisfaction were significantly lower than those of employers (P<0.05) . The overall satisfaction rate of workers with diagnosis of occupational diseases was lower than that of workers with health examination (P=0.011) . The recognition rates of workers with health examination on clear instructions process and attentions notification were lower than those of employers (P<0.016) . There was a significant difference in the overall satisfaction between respondents in different regions (P<0.01) . Conclusion The service of occupational health examination and occupational disease diagnosis services should be further improved. We should better learn the demands of employees and employers, improve service attitude, optimize service processes, shorten service time, and improve service quality and satisfaction.

Objective To explore the effect of multitargeted kinase inhibitor GNF-7 on the expression profiling of leukemia cell lines.Methods The expression profiles dataset GSE49534 was downloaded from the Gene Expression Omnibus (GEO) database.The BRB-Array Tools software package was employed to screen the differentially expressed genes (DEGs),then the Gene Ontology (GO) function,pathway enrichment,gene interaction network,and pathway relation network analyses were conducted based on these differential genes.Results Totally,847 differential genes were screened out,of which 426 genes were up-regulated and 419 genes were down-regulated.GO enrichment analysis showed that DEGs mainly performed the molecular functions of binding,protein kinase activity,and signal transducer activity,and participated in biological process of signal transduction,small molecule metabolic process,and apoptotic process.The pathway analysis found that DEGs were mostly enriched in ribosome biogenesis in eukaryotes,metabolic pathways,Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway,etc.The network analyses mining identified the hub genes,which included polyribonucleotide nucleotidyltransferase 1 (PNPT1),adenylate kinase 4 (AK4),Janus kinase 2 (JAK2),signal transducer and activator of transcription 2 (STAT2),MYC;and core pathways such as mitogen-activated protein kinase (MAPK) signaling pathway,apoptosis,cell cycle,pathways in cancer.Conclusion The GNF-7 inhibited leukemia cells via induction of apoptosis and cell cycle arrest.

Objective:To explore the effects of optimal nutritional guidance based on chronic kidney disease (CKD) integrative outpatient service in improving adherence to low protein diet and the nutritional status of non‐dialysis CKD patients . Methods :The 196 stage 3B to 4 CKD patients were randomly assigned to the optimized‐nutrition group and the traditional group .All participants received dietary counseling aimed at achieving a daily protein intake of 0 .6 g/kg and a daily energy intake of 30‐35 kcal/kg by two different modes of nutrition management . By using MDRD to calculate the eGFR , the nutritional status including human body measurement ,biochemical indexes and subjective global nutritional assessment (SGA ) were assessed every three months in one‐year follow‐up .Results:There was no significant difference in change of the eGFR between the two groups .Compared with the traditional group ,the optimized‐nutrition group had better low protein diet adherence ,lower dietary protein intake ,higher dietary calorie ,higher serum albumin level and weight ,lower serum urea nitrogen and parathyroid hormone ,and the nutritional status assessed by SGA improved significantly (P<0 .05) .Conclusions :The optimal nutritional guidance is effective in increasing patient adherence ,improving the nutrition status and with good safety .

Objective:To explore the mechanism of hypertension inducing erectile dysfunction(ED)using transcriptomics and network pharmacology.Methods:We randomly divided 12 male rats with spontaneous hypertension(SHT)into an L-arginine(LA)group(n=6)and an SHT model control(MC)group(n=6),took another 6 Wistar Kyoto male rats as normal controls(NC),and treated the animals in the LA group by intraperitoneal injection of LA at 400 mg/kg and those in the latter two groups with physio-logical saline,once a day,all for 7 days.Then we observed the blood pressure and penile erection of the rats,and determined the ex-pressions of the cGMP/PKG signaling pathway-related proteins and mRNAs in different groups using ELISA,Western blot and RT-qPCR.Results:Transcriptomics combined with network pharmacology showed that the cGMP/PKG signaling pathway played a key role in hypertension-induced ED.In vivo animal experiments revealed a significantly lower frequency of penile erections in the MC than in the NC group(1.33±0.52 vs 2.67±0.51,P＜0.05).The protein expressions of eNOS,PKG and sGC were markedly de-creased in the model controls compared with those the normal controls(P＜0.05),but remarkably upregulated in the LA group com-pared with those in the MC group(P＜0.05).Conclusion:Hypertension decreases the expressions of eNOS,NO,sGC,cGMP and PKG proteins and the level of testosterone by inhibiting the cGMP/PKG signaling pathway,which consequently suppresses the relaxa-tion of the penile vascular smooth muscle and reduces erectile function.

Background:Chronic kidney disease (CKD) with moderate-to-severe renal dysfunction usually exhibits an irreversible course,and available treatments for delaying the progression to end-stage renal disease are limited.This study aimed to assess the efficacy and safety of the traditional Chinese medicine,Niaoduqing particles,for delaying renal dysfunction in patients with stage 3b-4 CKD.Methods:The present study was a prospective,randomized,double-blind,placebo-controlled,multicenter clinical trial.From May 2013 to December 2013,300 CKD patients with an estimated glomerular filtration rate (eGFR) between 20 and 45 ml,min-1· 1.73 m-2,aged 18-70 years were recruited from 22 hospitals in 11 Chinese provinces.Patients were randomized in a 1∶1 ratio to either a test group,which was administered Niaoduqing particles 5 g thrice daily and 10 g before bedtime for 24 weeks,or a control group,which was administered a placebo using the same methods.The primary endpoints were changes in baseline serum creatinine (Scr) and eGFR after completion of treatment.The primary endpoints were analyzed using Student's t-test or Wilcoxon's rank-sum test.The present study reported results based on an intention-to-treat (ITT) analysis.Results:A total of 292 participants underwent the ITT analysis.At 24 weeks,the median (interquartile range) change in Scr was 1.1 (-13.0-24.1) and 11.7 (-2.6-42.9) μmol/L for the test and control groups,respectively (Z =2.642,P =0.008),and the median change in eGFR was-0.2 (-4.3-2.7) and-2.2 (-5.7-0.8) ml·min-1.1.73 m-2,respectively (Z =-2.408,P =0.016).There were no significant differences in adverse events between the groups.Conclusions:Niaoduqing particles safely and effectively delayed CKD progression in patients with stage 3b-4 CKD.This traditional Chinese medicine may be a promising alternative medication for patients with moderate-to-severe renal dysfunction.