Objective:To observe the effect of 5-aza-2'-deoxycytidine on proliferation and apoptosis of ovarian cancer cell lines(SKOV3 and 3AO)and on expression of mismatch repair(MMR)genes(hMLH1 and hMSH2)in SKOV3 and 3AO cells.Methods:Human ovarian cancer cell lines SKOV3 and 3AO were treated for 3 d with 5-aza-CdR(0.5,5,50 ?mol/L),a specific demethylation agent,and then cultured in RPMI 1640 medium for another 7 d.The cells growth was observed by MTT assay before and after 5-aza-CdR treatment and the cell apoptosis was analyzed by flow cytometry.The expression of hMLH1 and hMSH2 mRNA was examined by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR).Results:5-aza-CdR(0.5,5,50 ?mol/L)obviously inhibited the growth of SKOV3 and 3AO cells compared in a concentration dependent manner.The apoptosis rates of SKOV3 cells were(10.59?1.57)%,(17.52?1.72)%,(34.10?1.45)% after treated with 0.5,5,50 ?mol/L 5-aza-CdR,respectively;and the apoptosis rates of 3AO cells were(11.11?2.21)%,(17.24?1.11)%,and(26.53?2.00)%,respectively,which were all markedly higher than those of control group(P

Objective To understand the clinical features and the differences of age on children with asthma exacerbation.Methods Totally 395 inpatients with asthma exacerbation of our hospital were enrolled from January 2013 to December 2015.All the inpatients were divided into 2 groups:＜ 5 years of age group and ≥5 years of age group.The general data,epidemiology,predisposing factors,respiratory tract infection and pathogenic spectrum of these children were compared and analyzed.Results Average age of all inpatients was 4.25 ±2.51 years old,the proportion of male and female was 2.06:1,and autumn was the epidemical season.367 inpatients(92.91％)were complicated with respiratory tract infection.113 inpatients (28.61％) had been diagnosed with asthma before hospitalization.31 inpatients(7.89％) were followed up with long term standardized treatment.Compared with ≥ 5 years of age group,inpatients of ＜ 5 years of age group were more easily to be complicated with respiratory tract infection (x2 =9.856,P =0.007),especially with pneumonia (x2 =9.846,P =0.002).The detection rate of respiratory syncytial virus in ＜ 5 years of age group were higher than ≥ 5 years of age group (x2 =10.870,P =0.001).But there were no significant difference between the two groups in the detection rate of mycoplasma.(x2 =3.090,P =0.079).The diagnostic rate of asthma before hospitalization and rate of long term standardized follow-up treatment in ＜ 5 years of age group were lower than ≥5 years of age group (x2 =16.742,P =0.000;x2 =12.948,P =0.000).Conclusion Respiratory tract infection,non-timely diagnosis and standard treatment were major predisposing factors for asthma exacerbation of children.＜ 5years of age group were easily to be complicated with respiratory tract infection,especially with pneumonia and respiratory syncytial virus infection.The diagnostic rate of asthma before hospitalization and rate of long term standardized follow-up treatment were lower.

ObjectiveTo observe the effect of reverse acupuncture by stages combined with facilitation techniques on motor function of stroke patients.Methods114 stroke patient were randomly divided into the treatment group (n=68) and control group (n=46). The patients of the treatment group were treated with reverse acupuncture by stages combined with facilitation techniques. Those of the control group were treated with general acupuncture combined with facilitation techniques. The period of treatment of two group was 30 days.ResultsAfter treatment, the scores of motor function and activities of daily living of the treatment group were significantly higher than that of the control group ( P<0.01).ConclusionThe reverse acupuncture by stages combined with facilitation techniques can obviously improve motor function of stroke patients.

Objective To investigate the application effect of Kangfuxin Liquid on pain relief saliva SigA and IgG levels in patients with oral ulcer.Methods A total of 98 patients with oral ulcer treated in our hospital from January 2013 to July 2015 were randomly divided into two groups,with 49 cases in each group.The observation group was treated with Kangfuxin oral liquid,the control group was treated with stomatitis spray.The treatment effect, relieve pain, ulcer healing time and saliva SIgA, IgG levels in the two groups were observed,and whether the record of concurrent side effects.Results The effective rate of the treatment group was 89.79% higher than the control group 65.30%(P<0.05).Compared with before treatment, the observation group after treatment, the level of saliva SigA increased, IgG level decreased(P<0.05), the control group after treatment of saliva SIgA, IgG level of improvement is greater than that of the control group, the difference was statistically significant(P<0.05).The pain relief and healing time of the observation group were significantly lower than the control group(P<0.05).Conclusion The clinical effect of Kangfuxin liquid in treatment of oral ulcer in patients with significant clinical effect,reduce local ulcer pain, promote ulcer necrotic cell shedding and granulation tissue of newborn,increase the SIgA, IgG content, strengthen the local immunity response, promote oral ulcer healing, worthy of promotion.

AIM: To establish the quality standard for Chanhua Junsitifen Capsule(Paecilomyces cicadae(Miquel) samson). METHODS: Amino acids,adenine,adenosine,uridnine in Chanhua Junsitifen Capsule were identified by TLC and adenosine was determined by HPLC.Chromatographic conditions were adopted as follows:co-lumn: Inertsil C_18(4.6 mm?250 mm,5 ?m),mobile phase: MeOH-0.05 mol/L KH_2PO_4 solution(15∶85),wavelength: 260 nm,flow rate: 1.0 mL/min. RESULTS: The average recovery was 99.60 % and RSD was 0.62 % and the linear range of adenosine was in the range 0.007 475-2.99 ?g. CONCLUSION: This method is simple,rapid with a good reproducibility.This method can be used for the quality control of Chanhua Junsitifen Capsule.

We constructed plasmid pMTac to overexpress 3-ketosteroid-Δ1-dehydrogenase (KSDD) in Mycobacterium neoaurum JC-12 for improving androst-1,4-diene-3,17-dione (ADD) production. To construct pMTac, pACE promoter on pMF41 was replaced by tac promoter, and then four recombinants were constructed, which were M. neoaurum JC-12/pMF41-gfp, M. neoaurum JC-12/pMTac-gfp, M. neoaurum JC-12/pMF41-ksdd and M. neoaurum JC-12/pMTac-ksdd. Fluorescence detection results show that much more green fluorescent protein (GFP) was expressed in M. neoaurum JC-12/pMTac-ksdd than M. neoaurum JC-12/pMF41-ksdd. The activity of KSDD was 2.41 U/mg in M. neoaurum JC-12/pMTac-ksdd, 6.53-fold as that of M. neoaurum JC-12 and 4.36-fold as that of M. neoaurum JC-12/pMF41-ksdd. In shake flask fermentation, ADD production of M. neoaurum JC-12/pMTac-ksdd was 5.94 g/L, increased about 22.2% compared to the original strain M. neoaurum JC-12 and 12.7% to M. neoaurum JC-12/pMF41-ksdd. AD (4-androstene-3,17-dione) production of JC-12/pMTac-ksdd was 0.17 g/L, decreased 81.5% compared to M. neoaurum JC-12 and 71.2% to M neoaurum JC-12/pMF41-ksdd. In the 5 L fermenter, 20 g/L phytosterols was used as substrate, ADD production of M. neoaurum JC-12/pMTac-ksdd was improved to 10.28 g/L. pMTac is favorable for expressing KSDD in M. neoaurum JC-12, and overexpression of KSDD has beneficial effect on ADD producing, and it is the highest level ever reported using fermentation method in M. neoaurum.
Androstadienes ; metabolism ; Fermentation ; Industrial Microbiology ; Mycobacterium ; Oxidoreductases ; genetics ; metabolism ; Phytosterols ; metabolism ; Plasmids

For newly diagnosed multiple myeloma (MM) patients, bortizomib based on chemotherapy has achieved desirable complete remission rate, good progression-free survival and overall survival, which is now a first-line choice in both transplant-eligible and transplant-ineligible patients. In recent years, other new treatment options such as proteasome inhibitors , immunomodulators , monoclonal antibody and cellular immunotherapy have posed a challenge to the first-line therapeutic status of bortizomib because of their favorable efficacy and safety. Oral proteasome inhibitor or monoclonal antibody combined with immunomodulators and dexamethasone might become the first-line induction therapy of newly-diagnosed MM patients. Moreover, chimeric antigen receptor T-cell immunotherapy (CAR-T) may also play a role in the first-line treatment, especially for the induction therapy of high-risk patients and the consolidation treatment of the elimination in minimal residual disease.

Aim To investigate the effects of dopamine(DA)on insulin secretion from rat islets and the possible mechanism.Methods Pancreatic islets were obtained from the pancreatic of male SD rats by collagenase P digestion and histopaque-1077 density gradient separation.Insulin secretion experiment was used to observe the change of insulin release after DA treatments.As to study the potential mechanisms of the effects of DA,patch-clamp experiment and calcuim image technique were applied to test the depolarization-evoked Ca2+ currents,action potential duration and intracellular Ca2+ concentration.Results In 2.8 mmol·L-1 glucose,DA had no effect on insulin secretion;in 16.7 mmol·L-1 glucose,dopamine inhibited insulin secretion in a dose-dependent manner.DA inhibited the inward calcium current,shorten the action potential duration,and reduced the intracellular Ca2+ concentration.Conclusion DA inhibits insulin secretion maybe by decreasing the inward calcium current leading to shorten the action potential duration and reduce the intracellular Ca2+ concentration.

Objective To study the immune tolerance effect after renal transplantation,an animal model of tolerant dentritic cell adaptive transfer renal transplantation was constructed.Methods Tolerant Dentritic cells from SD rats were separated and purification in vitro,after a series of procedures,the Dentritic cells were administrated in the portal vein of Wistar rats,then conduct the operation of renal transplantation for Wistar rats.Renal tissue pathology and splenic lymphocyte reaction were test in 3、5、7、9、11、和13d after renal transplantation.Results After the renal transplantation,time of acute rejection happened in control group and experimental group is 4.69?1.26d and 10.69?1.63d,respectively.The mean survive time between two group has significant difference.Conclusion Renal transplantation immune tolerance could be induced by the tolerant dentritic cell of donor,by the mean of administrating the cell into host portal vein.The immune tolerance has specificity.

Objective To investigate the clinicopathological features and prognosis in patients with thymoma and patients with thymoma and myasthenia gravis ( MG) . Methods A retrospective analysis was performed on the clinicopathological data of 161 patients pathologically diagnosed with thymoma alone or thymoma and MG from 2008 to 2014. In those patients, 128 had thymoma with MG and 33 had thymoma alone. The survival rates were calculated using the Kaplan?Meier method and analyzed using χ2 test or Fisher′s exact probability test. Results The mean age of onset was 45. 2 years for patients with thymoma and MG and 48. 5 years for patients with thymoma alone. In patients with thymoma and MG, 74. 2% had a tumor diameter of ≤5 cm, while 75. 8% of patients with thymoma alone had a tumor diameter of ≥5 cm. According to the Masaoka staging system, 78. 1% of patients with thymoma and MG had stage Ⅰ+Ⅱdisease, while 51. 1% of patients with thymoma alone had stage Ⅲ+Ⅳ disease. There was no significant difference in the 3?year overall survival ( OS) rate between the two groups ( 98. 1% vs. 81. 8%, P=1. 000) . The 5?year OS rate was significantly higher in patients with thymoma and MG than in patients with thymoma alone ( 91. 1% vs. 42. 9%, P= 0. 000 ) . In all patients, 140 patients with complete resection had significantly higher 3?and 5?year OS rates than 21 patients with incomplete resection ( 97. 2% vs. 58. 8%, P=0. 000;92. 7% vs. 25. 0%, P=0. 000). In patients with stage Ⅱ disease, there were no significant differences in the 3?or 5?year OS rates between patients with complete resection alone ( n=25) and patients with complete resection and postoperative radiotherapy ( n=25) ( 95% vs. 100%, P=1. 000;86% vs. 100%, P=0. 467). Conclusions Compared with patients with thymoma alone, patients with thymoma and MG have an earlier age of onset, substantially smaller tumor diameters, and earlier Masaoka stages. MG and complete resection are positive prognostic factors for patients with thymoma. Radiotherapy after complete resection can reduce the recurrence rate in patients with stage Ⅱ disease.