Objective To evaluate the effect of oxycodone postconditioning on myocardial ischemia-reperfusion (I/R) injury in rats.Methods Forty pathogen-free healthy adult male Sprague-Dawley rats,weighing 200-300 g,were randomly divided into 4 groups (n =10 each) using a random number table:sham operation group (group S),myocardial I/R group (group Ⅰ),oxycodone postconditioning group (group O),and selective protein kinase C inhibitor chelerythrine group (group CH).Myocardial ischemia was induced by 30 min occlusion of the left anterior descending branch of the coronary artery,followed by 120 min reperfusion.In group S,the left anterior descending branch of the coronary artery was only exposed but not ligated.In group CH,chelerythrine 5 mg/kg was injected intravenously and slowly via the jugular vein before ligation which was performed immediately after administration.In O and CH groups,oxycodone 0.5 mg/kg was injected intravenously and slowly via the jugular vein at 2 min before reperfusion.Arterial blood samples were taken at 120 min of reperfusion to detect the levels of cardiac troponin Ⅰ (cTnI) and creatine kinase-MB (CK-MB) in serum.The hearts were removed after the animals were sacrificed to measure the myocardial infarct size by TTC staining.Results Compared with group S,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly increased in Ⅰ,O and CH groups (P＜0.05).Compared with group Ⅰ,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly decreased in O and CH groups (P＜0.05).Compared with group O,the levels of cTnI and CK-MB in serum and myocardial infarct size were significantly increased in group CH (P＜0.05).Conclusion Oxycodone postconditioning can mitigate myocardial I/R injury in rats,and the mechanism is partially related to activation of protein kinase C signaling pathway.

Objective To investigate the effects of sinomenine on hind limb ischemia?reperfusion ( I∕R) injury and expression of Bcl?2 and Bax in skeletal muscle cells of rats. Methods Fifty?four healthy adult male Wistar rats, aged 6-8 weeks, weighing 180-220 g, were divided into 3 groups ( n=18 each) using a random number table: sham operation group ( group S) , group I∕R and sinomenine group ( group SIN) . The rats were subjected to 4 h of ischemia on the proximal part of the right hind limb using elastic rubber bands followed by reperfusion in I∕R and SIN groups. Sinomenine 60 mg∕kg was injected intraperito?neally at 30 min before reperfusion in group SIN, and the equal volume of normal saline was given instead of sinomenine at 30 min before reperfusion in S and I∕R groups. Immediately after onset of reperfusion and at 4 and 24 h of reperfusion, blood samples were collected from the cardiac apex to measure the concentra?tions of serum lactate dehydrogenase ( LDH) and creatine kinase ( CK) . The animals were sacrificed imme?diately after blood sampling, and the gastrocnemius specimens of the hind limb were immediately removed for determination of the wet to dry weight ratio ( W∕D ratio) and expression of Bcl?2 and Bax in gastrocnemi?us cells ( by immunohistochemistry) and for examination of the pathological changes after haematoxylin and eosin staining. The Bcl?2∕Bax ratio was calculated. Results Compared with group S, the gastrocnemius W∕D ratio and concentrations of serum LDH and CK were significantly increased, the expression of Bcl?2 was significantly down?regulated, the expression of Bax was significantly up?regulated, and the Bcl?2∕Bax
ratio was significantly decreased in I∕R and SIN groups ( P<0?05) . Compared with group I∕R, the gastroc?nemius W∕D ratio and concentrations of serum LDH and CK were significantly decreased, the expression of Bcl?2 was significantly up?regulated, the expression of Bax was significantly down?regulated, and the Bcl?2∕Bax ratio was significantly increased in group SIN ( P<0?05) . The pathological changes of the gastrocne?mius were significantly attenuated in group SIN as compared with group I∕R. Conclusion Sinomenine can attenuate hind limb I∕R injury, and the mechanism may be related to maintenance of the balance between Bcl?2 and Bax and to inhibition of apoptosis in skeletal muscle cells of rats.

Introduction: Steven-Johnson syndrome and Toxic Epidermal Necrolysis are rare but life threatening severe cutaneous adverse reactions to drugs. To determine the epidemiology of SJS, TEN and SJS/TEN overlap in University Malaya Medical Centre (UMMC). Methods: All patients admitted to UMMC from year 2013-2015 for SJS, SJS/TEN, TEN were recruited. The classification of SJS, SJS/TEN overlap and TEN was made based on the criteria laid down by Bastuji et al.2 Results: A total of 32 patients were recorded to have SJS, SJS/TEN overlap and TEN from 2013 to 2015. Drugs (n=32, 86.49%) remained the most common aetiology of SJS and TEN. The top three commonest drugs are allopurinol (n=6), followed by carbamazepine (n=5) and bactrim (n=3). Conclusion: This study demonstrates that drugs were the most common cause of SJS/TEN. Antibiotics were the most common drug group that caused SJS/TEN. Awareness of the common etiology such as drug is important and high index of suspicion of SJS and TEN is needed if patients were on the above medications.

Objective To investigate the cytotoxicity of vascular endothelial growth factor (VEGF) siRNA combined with fusion suicide gene yCDglyTK on human gastric cancer cell line in vitro.MethodsThe gastric cancer cell line SGC7901 was transfeeted with blank plasmid pcDNA3.1 (-) null [pcDNA3.1 (-) group], or VEGF-siRNA expression plasmid pGenesil-shVEGF (SGC7901/shVEGF group),or fusion suicide gene plasmid pcDNA3.1 (-)CV-yCDglyTK (SGC7901/CDTK group),or combined gene plasmid pcDNA3.1 (-)shVEGF-yCDglyTK (SGC7901//shVEGF-CDTK group) with calcium phosphate nanoparticles (CPNPs).Un-transfected gastric cells were set as control group.The stable transfected cells were selected by G418.The target gene expression was verified by RT-PCR and Western-blot.After given prodrug 5-fluorocytosine (5-FC), the biologic characters variation, apoptotic morphology and apoptotic rate of cells in each group were observed through cell growth curve by MTT assays, by-stander effect, Hoechst 33258 staining and flow cytometry.The data was analyzed with SPSS 13.0 software and multiple groups’ comparison was analyzed with LSD test.ResultsFour gastric cancer cells lines transfected with different plasmids were successfully established.The expression of gene yCDglyTK was detected both in SGC7901/CDTK cells and SGC7901/shVEGF-CDTK cells.By MTT assays, the cell growth curve indicated that the A570 value of SGC7901/shVEGF cells, SGC7901/CDTK cells and SGC7901/shVEGF-CDTK cells decreased significantly compared with that of SGC7901 and SGC7901/null cells after a 24-hour 5-FC treatment (P＜0.01).When the percentage of stable gene trasfected SGC7901 cells was 60％, 80％and 100％, the cell relative viability was 13.09％±2.40％, 9.74％±2.83％ and 5.68％±1.03％,respectively. A large number of cells in SGC7901/CDTK and SGC7901/shVEGF-CDTK group appeared typical apoptotic morphology under fluorescence microscope.The result of flow cytometry showed that the apoptosis rates in SGC7901/shVEG group、 SGC7901/CDTK group and SGC7901/shVEGF-CDTK group were 16.40％ ±4.68％, 57.63％ ± 4.96％ and 69.07％ ± 4.69％,respectively, and there were significant differences compared with control (P＜0.01).Conclusion VEGF siRNA combined with suicide gene can effectively kill gastric cancer cell line SGC7901.Apoptosis induction may be one of the important mechanisms of killing tumor cells.

Objective To explore how to reduce the incidence of pelvic mass after hysterectomy,and to evaluate clinical characteristics and the risks.Methods A retrospective study was carried out in 85 patients who returned for surgery due to a pelvic mass after prior hysterectomy for benign disease at Peking Union Medical College Hospital from January 2011 to June 2016.Results The majority of pelvic masses arising after hysterectomy and requiring surgery were benign (74％,63/85),while 19％ (16/85) were malignant and 7％ (6/85) were borderline.The most common type was ovarian endometrioma (24％,20/85) which usually occurs within the 5 years (16 cases),however,ovarian tumors (18 cases) were more likely to occur ≥10 years after hysterectomy.Characteristics associated with significantly increased likelihood of ovarian endometrioma were mainly ascribed to younger age [(47±5) years old],prior presence of endometriosis or adenomyosis (65％,13/20) and shorter time to pelvic mass onset [(3 ±3) years],as opposed to ovarian tumors (all P＜0.01).Additionally,higher number of prior abdominal surgeries significantly intensified the risk (RR=9.410,95％ CI:1.099-80.564,P=0.041).Conclusions The occurrence of pelvic mass after hysterectomy is tightly related to prior histologic findings,and particularly for ovarian endometrioma.Higher number of prior abdominal surgery will exacerbated the risk.It is effective to prevent the pelvic mass in women after hysterectomy if treat patients for the purpose of the risk factors.

Objective To evaluate different postoperative medications as maintenance treatment for rectovaginal endometriosis (RVE) patients after conservative surgery. Methods RVE patients who underwent transvaginal partial excision from January 2007 to September 2016 with regular outpatient follow-up were retrospectively screened. Those followed by a levonorgestrel-releasing intrauterine system (LNG-IUS) insertion or oral contraceptive drospirenone/ethinylestradiol (DRSP/EE) 3 mg/30μg administration were enrolled. Variations in endometriosis-related pain, sexual function and quality of life were measured by visual analogue scale (VAS), female sexual function index (FSFI) and short form 36-item health survey (SF-36) respectively. Results There were a total of 102 RVE patients with 48 (47.1%, 48/102) in LNG-IUS group and 54 (52.9%, 54/102) in DRSP/EE group included. A rapid and marked improvement was observed after 3 months postoperative medical treatment compared to preoperative in both groups (P＜0.01). In dysmenorrhea, for LNG-IUS group (2.5±0.8) versus (7.6±1.3;P＜0.01), for DRSP/EE group (2.7±0.6) versus (7.7 ± 1.4;P＜0.01);in FSFI, for LNG-IUS group (23.5 ± 2.0) versus (21.0 ± 2.7;P＜0.01), for DRSP/EE group (23.4 ± 1.2) versus (21.5 ± 2.2; P＜0.01); in SF-36, both groups had obvious improvements in physical component summary and mental component summary (P＜0.01), for LNG-IUS group (74±13) versus (56±19), (75±13) versus (55±17), for DRSP/EE group (73±11) versus (59±15), (75±9) versus (54±14). These effects were maintained stably and progressively during postoperative medication at 6-, 12-, 24-month follow up. Conclusion Transvaginal partial excision combined postoperative LNG-IUS or DRSP/EE treatment is a safe and viable technique to alleviate pain, improve sexual function and quality of life.

Chickenpox may lead to several neurological complications. Optic neuritis is one of the complications which has rarely been described, especially in immunocompetent individuals. We report a case of an 11-year-old immunocompetent girl who presented with sudden onset bilateral vision loss three weeks after varicella eruption. Ophthalmic examination revealed bilateral optic disc edema. Diagnosis of bilateral optic neuritis secondary to varicella was established based upon the preceding medical history, supported with clinical and radiological fndings.

To study the chemical constituents from the the deep-sea fungus Alternaria sp. F49. Seven compounds were isolated from the EtOAc extract by using silica gel, Sephadex LH-20, ODS and HPLC methods. Based on the spectroscopic analysis, their structures were identified as (8R)-5-O-methyl-orcinotriol (1), orcinotriol (2), α-acetylorcinol (3), 3'-hydroxyalternariol 5-O-methyl ether (4), altenusiol (5), altenusin (6), and 5'-methoxy-6-methyl-biphenyl-3,4,3'-triol (7). (8R)-5-O-Methyl-orcinotriol (1) is a new phenolic compound which has never been reported in the literature. Compounds 4-7 showed strong DPPH free radical scavenging activity; whereas compounds 1-7 showed strong ABTS free radical scavenging activity.

Objective To study the effects of Qilongtian on interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)in hypoxic pulmonary hypertension (HPH)rats.Methods All male rats were ran-domly divided into six groups:normal group,model group,bosentan group,Qilongtian high-,mid-and low-group.Except the rats in normal group,those in other five groups were under atmospheric hypoxia to establish HPH model.From the first day of modeling,all medication groups were i.g corresponding drugs for consecutive three weeks.Pulmonary hemodynamic parameters were detected by right heart catheteriza-tion under direct vision,mRNA expression of IL-1βand TNF-αin lung tissue were measured using ELISA,protein expression of IL-1βand TNFαin lung tissue were measured using immunohistochemistri-cal methods.Results Qilongtian and bosentan significantly reduced the level of mean pulmonary arterial pressure (MPAP),IL-1βand TNF-αexpression in HPH rats,and the effect of Qilongtian high dosage was stronger with the dose-dependence.Compared with normal group,expression of IL-1βand TNF-αmRNA and protein increased,and those in model group and bosentan group did significantly(P <0.01). Compared with the model group,there was no significant difference in bosentan group (P >0.05),the expression level of IL-1βand TNF-αwere significantly downregulated in all Qilongtian groups (P <0.01 or P <0.05).Compared with the bosentan group and all the Qilongtian groups,there were significant differences,and the Qilongtian high-,mid-dose group had more impact (P <0.01 or P <0.05).Con-clusion High,middle and low dosage of Qilongtian reduced pulmonary hypertension,with high dosage prior to bosentan.The mechanism of lower blood pressure may be via downregulating the expression of IL-1βand TNF-α,but it would be influenced by other vascular tension factors and angiogenic factors.

OBJECTIVETo investigate the effectiveness of human insulin-like growth factor I (hIGF-I) gene transferred into the cultured goat bone marrow mesenchymal stem cells with liposome, and find a new method of cell-mediated gene therapy.

METHODSBone marrow was extracted from adult goats and cultured in vitro by monolayer. Then the recombinant eukaryotic expression plasmid pIRES2-EGFP-hIGF was transfected into cells by FuGene 6. After transfection, the marker gene coding enhanced green fluorescent protein (EGFP) was observed at different time points. The hIGF concentration in the supernatant fluids was measured by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry stain of hIGF was performed to detect the target protein. Besides, reverse transcription polymerase chain reaction and flow cytometry were also adopted in order to find out the changes of cells after transfection.

RESULTSBone marrow stem cells were all in the long shuttle-like shape and adhered to the disk. The expression of EGFP was first found at 4 h after transfection. The amount and intensity of EGFP increased gradually during the period of detection and got to the peak degree at 72 h, after that decreased slowly. EGFP was also seen in the second generation cells, but the intensity was relatively faint. The IGF-I concentration secreted into the supernatant was in accordance with the EGFP observed with the peak concentration at 34.75 ng/ml. The outcome of RT-PCR and immunohistochemistry was positive. The morphology of many stem cells was changed into triangular or irregular forms under the circumstance of the secreted hIGF-I. Percentage of stem cells in the S stage increased after transfection.

CONCLUSIONThe hIGF-I gene can be transfected efficiently and safely into BMSCs by FuGene 6, and the hIGF-I protein can be secreted into the supernatant in a relatively high level during a long period, therefore accelerate the proliferation and differentiation of the transfected cells.

Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Genetic Vectors ; Goats ; Humans ; In Vitro Techniques ; Insulin-Like Growth Factor I ; biosynthesis ; genetics ; Male ; Mesenchymal Stromal Cells ; cytology ; Plasmids ; genetics ; Transfection