Adolescent ; Antibody Formation ; drug effects ; Asthma ; drug therapy ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Powders ; Th1 Cells ; drug effects ; immunology ; Th2 Cells ; drug effects ; immunology

Objective To discuss the significance of the application of real-time quantitative PCR(RQ PCR)for diagnosis and guidance therapy of cytomegalovirus(CMV)infection after allo- hematopoietic stem cell transplantation(allo-HSCT).Methods Thirty-three patients were undergo- ing allo-HSCT.After hematopoietic reconstitution,patients'peripheral blood samples were detected for CMV DNA by RQ-PCR periodically.Anti-viral therapy was begun,attenuated and stopped ac- cording to the results of CMV DNA detection.The effects of anti-viral therapy and patients' clinical results were observed.Results CMV DNA was detected in blood samples from 13 of 33 patients. There were 21 episodes in these patients.Only 1 episode wasn't controlled hecause the patient gave up the therapy.CMV DNA copies were disappeared soon or decreased and then disappeared during anti-viral therapy in the others'.Patients which had symptoms and/or dysfunction of organs were cured too.Each course of anti-viral therapy was shorter than ordinary course.Conclusions CMV in- fection can he diagnosed as early as possible by RQ PCR.To begin,attenuate and stop anti-viral ther- apy according to the results of RQ-PCR is safe.The course is shortened and the side effects of anti vi- ral drugs were attenuated.

BACKGROUND:Studies have reported that taurine has a certain therapeutic effect on the disease of various systems, such as nervous system, cardiovascular system, immune system and digestive system. The liver is the main place, also the important target organ, of taurine metabolism. Therefore, the relationship between taurine and hepatopathy has become a hot topic in recent years. OBJECTIVE:To investigate the influence of taurine on superoxide dismutase and malondialdehyde expression in the liver tissue of rat models of liver fibrosis induced by carbon tetrachloride. METHODS:Thirty male C57B/L rats of SPF grade were randomly and evenly divided into blank control, model and taurine groups. Rats in the blank control group were intraperitonealy injected with 100% peanut oil of 1 mL/kg, twice a week, in total 10 weeks. Rats in the model group were intraperitonealy injected with peanut oil of 1 mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks. Rats in the taurine group were intraperitonealy injected with peanut oil of 1mL/kg containing 20% carbon tetrachloride, twice a week, in total 10 weeks, and were intragastricaly administered taurine of 500 mg/kg per day starting from the 3rd week til the 10th week. RESULTS AND CONCLUSION:Compared with the blank control group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly increased (P < 0.05), the level of superoxide dismutase in the liver tissue was lowered (P < 0.05), the level of malondialdehyde in liver tissue was significantly increased (P < 0.05), and liver index was increased (P < 0.05) in the model group. Pathological examination showed that there were necrosis of liver cels, fat vacuoles, fibrous tissue hyperplasia and inflammatory cel infiltration in the rats of the model group. Compared with the model group, the serum levels of hyaluronic acid, laminin, typeⅢ procolagen, typeⅣ colagen, alanine aminotransferase, aspartate aminotransferase were significantly lowered (P < 0.05), the level of superoxide dismutase in the liver tissue was significantly increased (P < 0.05), the level of malondialdehyde in the liver tissue was significantly lowered (P < 0.05), and liver index was significantly decreased (P < 0.05) in the taurine group. Pathological examination showed that there were no inflammatory cel infiltration, fat vacuoles, and fibrous tissue deposition in the liver tissue. The results indicate that taurine can decrease the contents of superoxide dismutase and malondialdehyde, and relieve the degree of liver fibrosis induced by carbon tetrachloridevia exerting its antioxidative effects.

Objective The aim of this study was to evaluate the efficacy and safety of the approach of the Nuss procedure for the correction of pectus excavatum in children without thoracoscopy.Methods From Oct 2007 and May 2009,48 patients with pectus excavatum underwent Nuss procedure.Among them 22 were done under the thoracoscopic guidance,and the other 26 in a non-thoracoscopic way,in which,a bilateral extrapleural tunnel to the edge of sternum was created using a blunt dissection via a bilateral thoracic skin incision.Without introducing the thoracoscopy into the thoracic cavity,a steel bar was inserted in the entirely extrapleural tunnel and turned as the standard Nuss procedure.Results All 48 patients recovered uneventfully.There were no postoperative deaths and serious complications.A single alloy steel bar(23-40 cm)was used in all patients.In the non-video-assisted extrapleural group(n=26),no pneumothorax occurred,the operating time(after anesthesia)ranged from 24～38 minutes[mean(25.4±2.6)mins],blood loss was minimal(range,5-10 ml),and the hospital stay was ranged from 3-6 days[mean(4.5±1.1)days].In the thoracoscopic group(n=22),the corresponding figures were 40 to 60 minutes[mean(53.5±3.4)mins)],10 to 15ml,5-8days[mean(7.0±2.2)days],respectively.No recurrent of the funnel chest occurred during the 3-18 months(median 10.4 monthes)of follow-up.The bar displacement occurred in 1 case 2 months after operation,which was replaced with satisfied result.Conclusion The non-thoracoscopic approach of the Nuss procedure is a safe and less traumatic procedure for the correction of pectus excavatum.

The characteristic fingerprint of conventional dairy Nanhanshuishi was established by X-ray diffraction (XRD), based on similarity of caculation on public peaks by MATLAB software, and the feasibility of new dairy technology of microwave method was explored between XRD and the dissolution rate in artificial simulation gastric juices. The result showed that similarity of shared peak in XRD of conventional dairy Nanhanshuishi was > 95%, This XRD characteristic fingerprint of conventional dairy Nanhanshuishi had strong specificity, could be used to provide a reference for identification and quality evaluation. This study also showed that the similarity of microware dairy products and conventional dairy products was good, and the sample of microwave 15 min was the best, and new dairy method by the microwave could replace the traditional method.
Animals ; Chemistry, Pharmaceutical ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Medicine, Tibetan Traditional ; Microwaves ; Milk ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; X-Ray Diffraction

We amplified genes encoding UDP-glucose dehydrogenase, ecohasB from Escherichia coli and spyhasB from Streptococcus pyogenes. Both ecohasB and spyhasB were inserted into T7 expression vector pRX2 to construct recombinant plasmids pRXEB and pRXSB, and to express in E. coli BL21(DE3). After nickel column purification of UDP-glucose dehydrogenases, the enzymes were characterized. The optimum reaction condition of spyHasB was at 30 °C and pH 10. The specific activity reached 12.2 U/mg under optimum condition. The optimum reaction condition of ecoHasB was at 30 °C and pH 9. Its specific activity reached 5.55 U/mg under optimum condition. The pmuhasA gene encoding hyaluronic acid synthase was amplified from Pasteurella multocida and ligated with ecohasB and spyhasB to construct the coexpression vectors pBPAEB and pBPASB, respectively. The co-expression vectors were transformed into E. coli BW25113. Hyaluronic acid (HA) was produced by biotransformation and the conditions were optimized. When recombinant strains were used to produce hyaluronic acid, the higher the activity of UDP-glucose dehydrogenase was, the better its stability was, and the higher the HA production could reach. Under the optimal conditions, the yields of HA produced by pBPAEB/BW25113 and pBPASB/BW25113 in shake flasks were 1.52 and 1.70 g/L, respectively, and the production increased more than 2-3 folds as previously reported.
Biotransformation ; Escherichia coli ; enzymology ; Genetic Vectors ; Glucuronosyltransferase ; genetics ; Hyaluronan Synthases ; Hyaluronic Acid ; metabolism ; Pasteurella multocida ; enzymology ; Streptococcus pyogenes ; enzymology ; Uridine Diphosphate Glucose Dehydrogenase ; metabolism

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Objective To evaluate the diagnostic value of ~(99)Tc~m-methoxyisobutylisonitrile (MIBI) SPECT scintigraphy combined Iocalizable CT in the localization of ectopic parathyroid glands in hyperparathyroidism.Methods Retrospective data of surgery,pathology and imaging were collected from 28 patients with hyperfunctioning ectopic parathyroid glands.All cases underwent CT studies.Twenty-five patients had ~(99)Tc~m-MIBI planar imaging first:SPECT scintigraphy combined localizable CT was performed for the patients with abnormal radionuclide foci immediately.The fusion images obtained after reconstruction showed the exact location of the ectopic foci.Operative histopathologic results were regarded as "gold standards".Presuming 4 parathyroid glands as normal findings,findings confirmed by operation and pathology were regarded as positive,otherwise negative.The results of CT and radionuclide imaging were compared by X~2-test of four-foId table.Results Twenty-eight ectopic parathyroid glands were found in 28 patients,all pathologically confirmed as adenomss.CT found 22 foci,of which 17 were true positive,5 false positive,11 false negative,and 79 true negative.~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT found 23 foci,no false positive,2 false negative,and 75 true negative.The results showed that the sensitivities were 61% (17/28),92%(23/25),specificities 94%(79/84),100%(75/75),accuracies 86%(96/112),98% (98/100),positive predictive values 77%(17/22),100%(23/23),and negative predictive values 88% (79190),97%(75/77),respectively,for CT and radionuclide imaging.~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT was therefore significantly higher than CT in sensitivity(X~2=6.98,P＜0.01),specificity (X~2=4.61,P＜0.05),accuracy (X~2=10.30,P＜0.01),positive predictive value(X~2=5.88,P＜0.05) and negative predictive value (X~2=5.36,P＜0.05).Conclusion ~(99)Tc~m-MIBI SPECT scintigraphy combined localizable CT is superior to CT alone in the localization of ectopic parathyroid glands in hyperparathyroidism,but false negative can be found in some patients.

OBJECTIVETo explore attenuation and mechanism of endoplasmic reticulum stress (ERS)-mediated hepatocyte apoptosis in rats with alcohol-induced liver injury by Qinggan Huoxue Recipe (QGHXR) and its disassembled formulas (Qinggan Recipe and Huoxue Recipe respectively).

METHODSA rat model of chronic alcoholic liver injury was successfully established using a compound reagent of alcohol, corn oil, and pyrazol. The modeled rats were randomly divided into the model group, the QGHXR group, the Qinggan Recipe (QGR) group, and the Huoxue Recipe group (HXR). The CCl4 control group and the normal control group were also set up. There were ten rats in each group. All rats of modeled groups were gastrogavaged with alcohol compound reagent every morning. Rats in the QGHXR group (at the daily dose of 9. 5 g/kg, QGR group (at the daily dose of 3.0 g/kg), and HXR group (at the daily dose of 6.5 g/kg) were administered with corresponding medicines by gastrogavage every afternoon. Equal volume of normal saline was given to rats of the model group by gastrogavage. CCl4 was intraperitoneally injected at the dose of 0.3 mL/kg to rats in the CCl4 control group, once per week. Normal saline was given to rats in the normal control group by gastrogavage. The treatment was lasted for two weeks. Pathological changes of the liver were observed by histopathology. Serum total homocysteine (tHCY) level was detected by ELISA. The hepatocyte apoptosis rate was detected using flow cytometry. The gene and protein expressions of eukaryotic translation initiation factor 2 alpha (elF-2alpha), phosphorylation elF-2alpha (pelF-2alpha), glucose-regulated protein 78 (GRP78), and Caspase-3 in the liver were examined using Real-time PCR and Westen blot respectively.

RESULTSCompared with the normal control group, typical pathological changes of chronic alcoholic liver injury such as steatosis, inflammation, and even fibrosis occurred in model rats. The hepatocyte apoptosis obviously increased, with the apoptosis rate reaching the five-fold of that in normal rats. Besides, early apoptosis dominated. The serum tHCY level significantly increased. The expressions of p-elF-2alpha, GRP78, and Caspase-3 protein obviously increased (P < 0.01). Expressions of GRP78 and Caspase-3 mRNA significantly increased (P < 0.05, P < 0.01). Compared with the model group, the degrees of the liver injury and the hepatocyte apoptosis in the QGHXR group, the QGR group, and the HXR group were significantly alleviated. The serum tHCY level was significantly lowered. The protein expressions of p-elF-2a, GRP78, and Caspase-3 obviously decreased (P < 0.01). mRNA expressions of GRP78 and Caspase-3 obviously decreased in the QGHXR group (P < 0.05, P < 0.01). Only GRP78 mRNA expression obviously decreased in the QGR group (P < 0.05).

CONCLUSIONQGHXR and its disassembled formulas could attenuate ERS-mediated hepatocyte apoptosis in alcohol-induced liver injury rats by lowering the serum tHCY level and expressions of ERS apoptosis correlated factors.

Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Endoplasmic Reticulum Stress ; drug effects ; Eukaryotic Initiation Factor-2 ; metabolism ; Heat-Shock Proteins ; metabolism ; Hepatocytes ; drug effects ; pathology ; Homocysteine ; blood ; Liver Diseases, Alcoholic ; blood ; drug therapy ; pathology ; Rats ; Rats, Sprague-Dawley

Objective To analyze retrospectively the clinico-pathological features that influencing the occurrence of anastomotie leakage after low anterior resection of rectal cancer, as well as its management and outcome of patients. Methods The data of 513 patients underwent anterior resection for low rectal cancer from june 1999 to June 2007 were reviewed. Results The incidence of anastomotic leakage was 4.5 %(23/ 513). 20 patients underwent conservative therapy,while 3 patients underwent reoperation, all patients were cured without ileostomy or colostomy.Conclusion The occurrence rate of anastomotie leakage is closely re- lated to the type of operation,the Dukes staging,Diabetes Mellitus and Diarrhea. Local irrigation via the drainage tube is the main strategic point to manage the leakage.