Objective To observe the changes of metallothionein (M T) in the rat flap with ischemia-reperfusion injury. Methods A ri ght lower abdominal island flap was created in Wistar rats . 16 rats were random ly divided into two groups: the control ( n =8) and the ischemia-reperfusion injury group ( n =8). The contents of MT and malondialdehyde (MDA) were assaye d by 109 Cd-hemoglobin saturation and thiobarbituric acid (TBA) methods. Th e activity of myeloperoxidase (MPO) was measured by colorimetry. Results At ischemia 8 h, reperfusion 12 h and 24 h, the level of MDA in rat i schemia-reperfusion injury group increased 41.7 %, 111.4 %, and 135.7 %, the ac tivity of MPO increased 72.1 %, 218.9 % and 296.0 %, the content of MT increased 42.6 %, 52.8 % and 102.3 %, respectively, compared with the control group ( P

Objective To establish the method of determination for rhein, emodin, chrysophanol and physcion in Niuhuang Qinghuo Pills. Methods Sample was hydrolyzed by 20% H2 SO4 , and anthraquinones were extracted by soxhlet extraction with acetone as solvent, and were determined by HPLC with Kromasil C18 as column, methanol-0.5%H3 PO4 (75:25,V/V) as mobile phase at the flow of 1.0 mL/min, 230 nm as the detection wavelength.Results The linear relationship of rhein, emodin, chrysophanol and physcion were 1.50-150, 0.950-95.0, 1.30-130 and 1.15-115 mg/L. The anthraquinones were seperated completely, the recovery of 4 anthraquinones were 98.14% -101.3%.Conclusion This method is simple, accurate, steady, and could be used for the quality control of Niuhuang Qinghuo Pills.

Objective: To extract and determine the flavonoids of hawthorn fruits and their antioxidative effect. Method: The ultrasonic method was used for extracting flavonoids. The content of flavonoids was determined by spectrophotometry, and the antioxidation effect was determined by flow-injection chemiluminescence.. Results and Conclution: The recovery rate was 96%～105% and the coefficient of variation was 0.14 % by spectrophotometry. Hawthorn fruits had high antioxidative effect, showing dose-response relation.

AIM: To study the effects of exogenous metallothionein (MT) and ZnCl 2-induced MT production on biological action of homocysteine(HCY)in vascular fibroblasts METHODS: -TdR, -Pro incorporation and LDH leakage were measured, the cellular viabilities were calculated by trypan blue exclusion test and the intracellular contents of MT were assayed by [ 109 Cd]-hemoglobin saturation method in cultured rat vascular fibroblasts. RESULTS: Proliferation, collagen production of vascular fibroblasts in HCY-treated group were significantly increased compared with control group in a concentration-depedant manner. HCY (500 ?mol/L) increased LDH leakage and decreased the cellular viabilities ( P

BACKGROUND:Primary culture in vitro of neurons plays an important role in the development, regeneration, signal transduction mechanisms, neuropharmacology and gene expressions of the nervous system.
OBJECTIVE:To establish a simple method for primary culture of high-purity cortical neurons in neonatal Sprague-Dawley rats.
METHODS:Cortical tissues were acquired from neonatal Sprague-Dawley rats born 1 day. In traditional experimental group, the whole cortex was removed;in improved experimental group, the cortical tissues, 2-3 mm thick on the brain surface were removed. Single cel suspensions were prepared after papain digestion and centrifugation and were then seeded onto 24-wel culture plates containing neuron solutions for primary culture (1×105 per wel ). Cel s were identified by neuronal specific markers MAP-2 and Tuj1 after 3-day culture. The number of neurons and neurite length were observed under inverted phase contrast microscope and recorded at 6, 24, 48 and 72 hours, 5 and 7 days of culture, resprctively.
RESULTS AND CONCLUSION:The cultured cel s expressing MAP-2 and Tuj1 were neurons that could be used in the fol owing experiments. The purity of neurons in the improved experimental group was 92%at 3 days, while only 51%in the traditional experimental group. Cel s in both two groups had attached to the wal presenting with smal processes at 6 hours, and a simple neural network formed at the 3rd day until dense neural networks could be found at the 5th day. To conclude, our culture method herein is simple and convenient, and can be used to produce neurons with high purity, which wil be helpful for the experimental studies on cortical neurons from Sprague-Dawley rats.

OBJECTIVETo study the pharmacokinetics and biodistribution of pur derivative nanopaticles 4ac in mice.

METHODAfter intravenous administration of 15 mg x kg(-1) of pur derivative 4ac, the plasma and tissue concentration was detected by HPLC. The pharmacokinetics parameters were calculated by the method of 3p97.

RESULTThe concentration time curves were conformed to two compartment model. The Re of 4ac nanopaticles was 21.18 times higher than suspension.

CONCLUSIONOur present study demonstrates that, compared to nanopaticles and suspension, nanopaticles significantly alter the pharmacokinetics in plasma and tissues targeting. Biodistribution of pur derivative 4ac nanopaticles in heart of mice was better than 4ac suspension. It will become a new drug for cardiovascular disease therapy.

Animals ; Female ; Isoflavones ; chemistry ; pharmacokinetics ; Male ; Mice ; Nanoparticles ; chemistry ; Plant Extracts ; chemistry ; pharmacokinetics ; Pueraria ; chemistry ; Random Allocation ; Tissue Distribution

Objective To investigate the value of the immunohistochemistry and Western blot in the diagnosis of the benign muscular dystrophy with abnormal dystrophin expression.Methods The medical histories and clinical manifestations of 4 patients were collected.In addition to routine histological and histochemical studies,expression of dystrophin in muscle fibets was observed by immunohistochemical reaction(dys-N,dys-R and dys-C)and Western blot to anti-dystrophin antibody.Results Two patients had muscular weakness while another 2 patients had only muscular pain and elevated creatine kinase blood levels without muscular weakness.Histochemical stains showed atrophy,hypertrophy and fiber splitting in 2 patients,while only variation in fiber size was presented in anothor 2 patients.One patient had no reaction for dys-N,but had immunostains for dys-C and dys-R in the sarcolemma of muscle fibers.Western blot confirmed that the band of dys-C and dys-R was partly deficient,and the band of dys-N was absent compared with control.Three patients had no reaction for dys-R,but had immunostains for dys-C and dys-N.Compared with control,Western blot confirmed that the band of dys-R was absent,and the band of dys-C and dys-N were truncated.Conclusion The immunohistochemistry is stained with three anti-dystrophin antibodies to avoid diagnostic errors.Western blot is essential to further determine the type of dystrophin protein.

Objective To analyze the clinical characteristics and assess the outcome of treatment for cervical cancer during pregnancy.Methods A cohort of 13 patients with cervical cancer diagnosed during pregnancy from January 2001 to September 2011 in Peking Union Medical College Hospital (PUMCH) was retrospectively studied.Clinical information,gestational age at diagnosis,treatment options and maternal and child outcomes were collected and analyzed.Results Thirteen patients out of 2030 cases of invasive cervical cancer were diagnosed during pregnancy with an incidence of 0.64％ (13/2030).The Mean gestational age at diagnosis of 13 patients is 21+6 weeks.Two cases were diagnosed during the first trimester,8 cases at second trimester and 3 cases at third trimester respectively.Vaginal bleeding during the pregnancy was main clinical manifestation presented in 8 patients and all thirteen cases were diagnosed by biopsy with pathological types of squamous cell carcinoma in 10 cases.The International Federation of Gynecology and Obstetrics (FIGO) stage was Ⅰ in eleven cases and stage Ⅱ in two cases.Six patients of them received treatment promptly after diagnosis.The other 7 patients had delayed treatment with mean diagnosis-treatment interval time of 65 days due to fertility reasons,who ended pregnancy by cesarean section at mean gestational age of 34+6 weeks,two of them received chemotherapy with cisplatin + fiuorouracil (PF)or cisplatin respectively before the end of the pregnancy,while the one with PF chemotherapy experienced neonatal death.The rest 6 neonatal outcomes were good.As follow-up of 13 cases:11 cases in stage Ⅰ received surgical treatment,and two of which had recurrence respectively,15 months and 7 months post surgery,and one case had died.One case of Stage Ⅱ patients died and one had recurrence after 53 months after radiotherapy.The recurrence rate in 13 cases was 3/13 and the mortality rate was 2/13.Conclusions Most cases of cervical cancer diagnosed during pregnancy were in early FIGO stage.For those patients diagnosed in late pregnancy with strong fertility demand,considering delayed treatment according to FIGO stage of the disease and fetus maturity is appropriate.Chemotherapy during pregnancy may cause neonatal complications.

OBJECTIVETo investigate the expression of microRNA-128a (miR-128a) and its role in hepatocellular carcinoma (HCC).

METHODSNineteen pairs of fresh surgical specimens of HCC and adjacent tissues were examined for miR-128a expression using qRT-PCR. A miR-128a mimics or inhibitor was transfected into HCC cells, and the cell viability was analyzed by MTT assay. RND3, one of the potential targets of miR-128a, was predicted by bioinformatics software and demonstrated by dual luciferase reporter assay. The expression of RND3 after transfection was detected using qRT-PCR and Western blotting, and the cell cycle-related proteins were determined with Western blotting.

RESULTSThe expression of miR-128a were significantly up-regulated in HCC tissues as compared to the adjacent tissues (P<0.05). In cultured HCC cells, miR-128a promoted the cell proliferation and resulted in down-regulated RND3 mRNA and protein expressions by targeting RND3' 3'UTR (P<0.05) and also in the down-regulation of cyclin B1, cyclin D1 and CDK4 protein expressions.

CONCLUSIONmiR-128a is up-regulated in HCC and promotes HCC cell proliferation by targeting RND3.

Carcinoma, Hepatocellular ; metabolism ; Cell Proliferation ; Cell Survival ; Down-Regulation ; Humans ; Liver Neoplasms ; metabolism ; MicroRNAs ; genetics ; metabolism ; RNA, Messenger ; Transcriptional Activation ; Tumor Cells, Cultured ; Up-Regulation ; rho GTP-Binding Proteins ; metabolism