Objective To explore the intervention effects of trichostatin A ( TSA ) on specialization of rat bone marrow mesenchymal stem cells ( MSCs ) .Methods The rat MSCs were isolated, cultured and purified by the whole bone marrow adherent method in vitro, with morphological observation.The third generation of MSCs were selected, directional induced to osteoblasts, and divided into the TSA low dose group (0.1μmol/L), middle dose group (1μmol/L) , the high dose group (10μmol/L) according to different drug concentrations, seting up blank control group at the same time.MSCs proliferation and cell growth curve of each group were drawn by MMT, the activity of alkaliphosphatase ( ALP) was detected, and the levels of corebinding factor α1 (Cbfα1), basic fibroblast growth factor (bFGF) and insulin-like growth factors-1 (IGF-1) mRNA were detected by RT-PCR. Results The trend of MSCs growth curves in each groups were similar, compared with control group, the growth curve of TSA low dose group had no significant change, the TSA middle dose and high dose significantly promoted the proliferation of MSCs (P<0.05).Compared with control group, ALP activity of TSA low-, middle-and high-dose group were significantly higher at 4th,5th,6th(P<0.05).The expression levels of Cbfα1, bFGF and IGF-1mRNA were significantly higher than those of control group, respectively (P<0.05).Conclusion TSA can significantly promote the rat bone marrow mesenchymal stem cells to differentiate into osteoblast, which is possibly associated with up-regulation of Cbfα1, bFGF and IGF-1mRNA level.

The second class is not only a useful supplement to the first class but also an effective way to improve the comprehensive quality and ability of students.Through the deliberate research training and laboratory work,the second class of scientific research facilitates the medical students' developing scientific thinking,expanding professional knowledge,enhancing the ability to do scientific research and cultivating the spirit of teamwork.

AIM: To explore the different inhibitory effect of arsenic trioxide (As_2O_3) on hepatocarcinoma cell growth in SMMC-7721 and BEL-7402 cell lines and its mechanism. METHODS: The cell culture and trypan blue staining were used to study the inhibitory effect of arsenic trioxide on cell growth, and the glutathione (GSH) contents in hepatocarcinoma cells treated with arsenic trioxide were detected. RESULTS: Arsenic trioxide inhibited the growth of BEL-7402 cells in a time and dose-dependent manner. The inhibitory effect was significant at a lower dose of 0.50 ?mol/L for 24 h, however, to SMMC-7721 cells, a higher dose of 2.00 ?mol/L for 96 h was needed. The inhibitory rate of arsenic trioxide (0.25-2.00 ?mol/L) on BEL-7402 cell growth was higher than that on SMMC-7721 cells. The content of GSH in SMMC-7721 cells was much higher than that in BEL-7402 cells [(50.8?5.2) (?mol/g) protein and (18.7?1.4) ?mol/g protein, respectively]. CONCLUSION: There was a significant difference in inhibition of hepatocarcinoma cell growth by arsenic trioxide between BEL-7402 and SMMC-7721 cell lines, the cause of which may be due to the difference in GSH content in BEL-7402 and SMMC-7721 cells. [

Objective To study the effects of methylene blue(MB), a soluble guanylate cyclase inhibitor , on intestinal perfusion and oxygenation in dogs with septic shock.Methods A septic shock model was established by lipopolysaccharide infusion.After resuscitation with normal saline infusion,2mg/kg MB was infused within 15min in seven dogs.Blood flow of the superior mesenteric artery (SMA) was measured respectively at baseline,post shock for 1h, after resuscitation and 30min after MB injection.Blood gas analysis was done , and oxygenation was calculated.Results Blood flow of the SMA was significantly decreased by 61 8%(P0 0 5 ) ,butO2 extrincreasedby 14 8% .Conclusions Septicshockindogsresultsinadecreaseinintestinalperfusionandanincreasein oxygenextraction ,whichcanbeimprovedbyMBinjection .Theintestinalperfusionispartlyincreasedbyvolumetherapy .

Objective To study the expression of GRHL-3 in diffuse large B-cell lymphoma (DLBCL) tissues and its clinical significance.Methods One hundred and sixty-eight pathology paraffin-embedded diffuse large B-cell lymphomas tissues were collected from January 2006 to September 2011.Immunohistochemistry was used to detect the expression of GRHL-3 protein in the above tissues.Results The positive expression rate of GRHL-3 protein in the GCB type tissues was higher than that in the non-GCB type tissues [84.87 ％(101/119) vs 14.29 ％ (7/49), P ＜ 0.01].Further analysis indicated that in the non-GCB type tissues,the positive expression rate of GRHL-3 in the latter stage group was significantly higher than that in the early stage group [90.00 ％ (63/70) vs 77.56 ％ (38/49), P ＜ 0.01].The positive expression rate of GRHL-3 in the lactatede hydrogenase increased group was significantly higher than that in the normal lactated hydrogenase [91.67 ％ (77/84) vs 68.57 ％ (24/35), P ＜ 0.01].The positive expression rate of GRHL-3 in the extranodal involvement status ≥ 2 group was significantly higher than that in the extranodal involvement status 0-1 group [96.29 ％ (26/27) vs 81.52 ％ (75/92), P ＜ 0.05].The positive expression rate of GRHL-3 in the IPI score 4-5 group was significantly higher than that in the IPI score 0-1 group [91.30 ％ (65/69) vs 66.67 ％ (18/27), P ＜ 0.01] and IPI score 2-3 group [91.30 ％ (65/69) vs 79.96 ％ (18/23), P ＜ 0.05].However, the expression of GRHL-3 had no correlation with the gender, age, and performance status of DLBCL.Conclusion The positive expression rate of GRHL-3 protein in the GCB type tissues is higher than that in the non-GCB type tissues.The positive expression rate of GRHL-3 in the DLBCL is correlated with the Ann Arbor stage, lactate dehydrogenase, extranodal involvement status and IPI score.

With the rise of the incidence of Crohn's disease,clinicians began to recognize and pay attention to this disease.There is no effective method to cure the Crohn's disease currently,and the complications of the Crohn's disease are numerous and complex.Therefore,early diagnosis and treatment are very important.Due to many factors,the problen of diagnostic delay is widespread.Female patients diagnosed younger than 40 years old,the presence of ileal disease,use of non-steroidal anti-inflammatory agents,smoking history,parental symptoms are risk factors of diagnostic delay of Crohn's disease.Delayed diagnosis might increase the risk of complications,especially intestinal stenosis and operation-related complications.Crohn's disease needs to be early diagnosed and treated to improve the prognosis of patients.

Objective To investigate the effect of salmeterol/fluticasone (SM/FP) on serum inflammatory factors in the treatment of bronchial asthma in children.Methods 80 children with bronchial asthma from January 2015 to October 2015 in department of pediatrics of first affiliated hospital of Zhejiang Chinese medicine university were selected and randomly divided into two groups.The control group were given routine clinical treatment, the experimental group were treated on the basis of the control group with salmeterol/fluticasone (SM/FP), for 4 weeks.The serum IL-2, IL-4, IFN-α, T-lymphocyte subsets and clinical efficacy between the two groups were compared.Results Compared with control group, the serum levels of IL-2 and IFN-γin experimental group were higher, IL-4 in experimental group was lower ( P <0.05 ); the serum CD3 +T, CD4 +T and CD4 +T /CD8 +T levels in experimental group were higher, the serum CD8 +T was lower than those in control group (P<0.05); the total efficiency of the experimental group (92.5%) was significantly higher than that of control group (75.0%) (P<0.05).Conclusion The salmeterol/fluticasone (SM/FP) has the good efficacy in the treatment of bronchial asthma in children, which could effectively regulate T lymphocyte subsets proportion and the level of cytokines.

Objective To investigate the effects of Tubastatin A Hcl,a selective HDAC6 inhibitor,on the development of chronic asthmatic mice with airway inflammation,airway remodeling and airway hyperresponsiveness. Methods BALB/C mice were randomly divided into control group, asthma group,dexamethasone group and Tubastatin A Hcl group. The airway resistance,total cells and different cells in BALF,IL?4,IL?5,TGF?β1 were detected by ELISA. HE、AB?PAS and Masson trichrome staining were carried out to assess the airway inflammation and remodeling. Immuno?histochemical staining and western blotting were adopted to determine the expression ofα?SMA and TGF?β1. Results After drugs treatment,air?way resistance decreased,and levels of IL?4,IL?5,TGF?β1,total inflammatory cells and eosinophils in BALF were relieved. Meanwhile,inflamma?tory cells infiltration,goblet cells metaplasia and collagen deposition in lung tissue were also reduced,but all of above the effects of dexamethasone were better than Tubastatin A Hcl. The expression ofα?SMA and TGF?β1 in the lung tissue decreased significantly after treatment ,in which Tu?bastatin A Hcl were slightly better than dexamethasone treatment. Conclusion Tubastatin A Hcl can effectively relieve airway inflammation ,air?way remodeling and airway hyperresponsiveness in chronic asthmatic mice ,but its effect of anti?inflammatory is worse than dexamethasone treat?ment,while it is better than dexamethasone in the effect of relief airway remodeling.

Objective:To analyze the utilization and tendency of oral hypoglycemic agents in some hospitals of Wuhan from 2010 to 2012. Methods:The relative data of oral hypoglycemic agents used in 32 hospitals of Wuhan area during 2010-2012 were analyzed using consumption sum, DDDs and defined daily dose as the indices. Results:The consumption sum of oral hypoglycemic agents was increased year after year;acarbose and gliclazide were widely used and occupied front places among the drugs. However, the percent-age of traditional Chinese medicine used for decreasing blood sugar was declined. Conclusion:During 2010-2012, the DDDs value and the consumption sum of oral hypoglycemic agents in Wuhan area are stable and normal, and the application will be further developed.

Objective To investigate the relationship between the expression of Toll-like receptor 4 (TLR4) on mononuclear cells in peripheral blood and changes of serum vitamin D level in children with bronchiolitis.Methods The children who were diagnosed as bronchiolitis and received treatment in the Second Affiliated Hospital of Harbin Medical University from October 2013 to January 2014 were chosen as the pre-treatment group,and then divided them into moderate group and severe group according to the clinical symptoms,20 cases for each group.Then the cases in pre-treatment group who recovered after treatment were recruited as the after-treatment group,and the children who were healthy and medical examination in the Second Affiliated Hospital of Harbin Medical University in the same period were recruited as the healthy control group.The expressions of TLR4 on CD14 labeled mononuclear cells in the periphera were measured by flow cytometry.The level of 25 (OH) D in serum was detected by enzyme linked immunosorbent assay (ELISA).Results (1) The expression level in children with bronchiolitis of TLR4:the mode-rate group [(18.98 ±2.29)％] and severe group [(30.13 ±2.13)％] increased significantly (P ＜0.05) compared with control group [(1.17 ± 0.57) ％].And the expression level of moderate group [(2.02 ± 0.48) ％] and severe group [(11.43 ± 1.52) ％] decreased significantly after treatment (P ＜0.05).(2) Serum vitamin D level in children with bronchiolitis of the moderate group[(17.16 ± 3.34) μg/L] and severe group [(6.56 ± 2.28) μg/L] were lower than healthy control group [(53.69 ± 20.18) μg/L] before treatment (P ＜ 0.05),especially the severe group [(6.56 ±2.28) μg/L].The level of moderate group [(9.59 ± 2.31) μg/L] and severe group [(4.70 ± 0.67) μg/L] became lower after treatment (P ＜ 0.05).(3) Both severe group (r =-0.491,P ＜ 0.05) before treatment and moderate group (r =-0.436,P ＜ 0.05) after treatment showed negative correlation between TLR4 on mononuclear cells in peripheral blood and serum 25 (OH)D level in children with bronchiolitis.And no correlation was found among healthy control group,moderate group before treatment and severe group after treatment (P ＞ 0.05).Conclusions The conditions of children with bronchiolitis was positively correlated with the expression level of TLR4,and negatively correlated with the vitamin D level.The serum 25 (OH) D decreased steadily during the treatment.The expression of TLR4 in monocytes has a certain correlation with the level of vitamin D in children with bronchiolitis.