Objective: To study the influence of inhibitor of differentiation 2 (Id2) on cell proliferation and migration in SKOV3 cells, and to investigate the influence of HLH domain deletion in 1d2 gene on SKOV3 cells. Methods: Human ovarian cancer cell line SKOV3 was transfected with pcDNA3.1-Id2, pcDNA3. 1-Id2-DBM and pcDNA3. 1-Id2-DBM-δHLH vectors by SuperFect Transfection Reagent. The cells transfected with blank vector pCDNA3. 1 were used as control. Id2 protein was detected by Western blot, and mRNA transcriptions of Id2, Id2-DBM (D-box mutant), and Id2-DBM-δHLH were measured by RT-PCR. Cell proliferation was assessed by MTT assay. Cell invasion was detected by scratch test and transwell chamber assay. The regulatory effect of Id2 on E-cadherin was evaluated by Western blot. Results: The mRNA and protein levels indicated that the plasmid was successfully transfected. Cell growth curve suggested that the proliferation of cells had no significant difference between each group. Compared with the control group, cell migration ability was significantly enhanced after transfection with pcDNA3. 1-Id2, pcDNA3. 1-Id2-DBM and pcDNA3. 1-Id2-DBM-δHLH vectors. The downregulation of E-cadherin protein was accompanied with increased migration capability in pcDNA3. 1-Id2-DBM and pcDNA3. 1-Id2-DBM-δHLH vectors-transfected cells. Conclusion: Overexpression of Id2 promotes the migration capability of SKOV3 cells, which might be related with the downregulation of E-cadherin. The action still exists after the HLH domain deletion in Id2 gene.

Objective:To investigate the causes, characteristics and treatment of maxillofacial trauma in children. Methods: A retrospective analysis of 470 consecutive maxillofacial records of the patients not older than 14 years was conducted. Data regarding age, gender, cause, anatomic site and treatment were reviewed. Results:Most of the patients were mals(335 cases, 71.3% ), with a male and female ratio of approximately 2. 5;1 of the injuries, 28. 5% were due to accidental falls, 17.0% traffic accidents and 10.0% sharp implementt cutting. Injuries of soft tissue often occurred on gingiva, cheek, lip and chin. Mandibuir fratures were the most common (55.1% ) of all bone fractures. Conservative therapy, such as closed reduction surgery, maxillomandibular fixation, was usually performed. Conclusion: ①Boys are more tendent to be victims of maxillofacial trauma than girls. ②Falls are the first cause of child victims. ③Gingiva, chin, lip, check and mandible are the most commonly injuried sites.

OBJECTIVETo observe the effect of Jianpi Bushen Qingchang Huashi Recipe (JBQHR) on proliferation and migration of bone marrow mesenchymal stem cells (BMSCs).

METHODSBMSCs were isolated and cultured in vitro with adherence screening method to prepare cell suspension. No drug intervention was given to BMSCs in the vehicle control group. JBQHR at 0.39, 0.78, 1.56 µg/mL was added in BMSCs of low, mid, and high dose JBQHR groups for co-incubation. Its effect on the proliferation of BMSCs was detected by CCK-8. BMSCs migration and chemotactic ability was detected using Transwell method. Each dose JBQHR combined ERK kinase inhibitor U0126 was set up as control. The phosphorylation of extracellular regulated protein kinase (ERK) and CAMP responsive element-binding protein (CREB) were detected by Western blot.

RESULTSCompared with the vehicle control group, the proliferation of BMSCs and BMSCs migration number could be promoted in the 3 JBQHR groups (P < 0.05). Besides, the proliferation of BMSCs was better in mid and high dose JBQHR groups than in the low dose JBQHR group (P < 0.05). Compared with the vehicle control group, the phosphorylation of ERK and CREB could be elevated in the 3 JBQHR groups (P < 0.05), and could be inhibited by U0126 (P < 0.01). Compared with the low dose JBQHR group, the phosphorylation of ERK increased in mid and high dose JBQHR groups with statistical difference (P < 0.05).

CONCLUSIONJBQHR could promote the proliferation and migration of BMSCs, and its mechanism might be related to ERK/CREB signaling pathway

Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cyclic AMP Response Element-Binding Protein ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Extracellular Signal-Regulated MAP Kinases ; metabolism ; Humans ; MAP Kinase Signaling System ; Mesenchymal Stromal Cells ; cytology ; drug effects

Objective To investigate the dynamic expressions of exchange protein directly activated by cyclic adenosine monophosphate (cAMP) (Epac) in rat model of hepatic fibrosis(HF).Methods Forty-two male SD rats were divided into control group (n = 6) and model group (n = 36)which was divided into six subgroups of day 4, week 1, week 2, week 4,week 6 and week 8 with six rats in each subgroup. The rat model of HF was established by intraperitoneal injection of dimethylnitrosamine (DMN). The pathological changes of liver were observed by Hematoxylin-Eosin and Masson staining. Reverse transcription-polymerase chain reaction (RT-PCR),immunohistochemistry and Western blot were employed to detect the mRNA and protein expressions of Epac1, Epac2 and transforming gronth factor (TGF)β1 during the process of modeling and localization in the liver. The statistical analysis was done using one-factor ANOVA, LSD-t test,Dunnett T3 test and Pearson linear correlation analysis. Results Rat model of liver fibrosis was established successfully. In control group, Epac1 (0. 031 28±0. 008 96) and Epac2 protein (0.034 43±0. 002 45) mainly expressed in the cytoplasm of hepatocytes. In model group, the level of Epac1 decreased at day 4 (0. 023 97±0. 003 81) and week 1 (0. 015 81±0. 002 48) ,then began to increase at week 2 of modeling and peaked at week 6 (0. 039 54±0. 001 43), which had statistical significance compared to the control group (t= 5.47,11.58 and - 6.18, respectively; all P＜0.05). Epac2 protein expression declined after modeling, reached the lowest level at week 4 (0. 011 21 ±0. 001 32), which had statistical significance compared to the control group (t= 24. 50, P＜0. 05). TGFβ1 protein expression increased after modeling and peaked at week 4 (0. 011 30±0.001 03) which had statistical significance (t= -23. 36, P＜0. 05) compared to the control group (0. 002 08 ±0. 000 18). The expressions of Epac1, Epac2 and TGFβ1 mRNA were consistent with the trend of protein levels.Correlation analysis showed that Epac1 protein was positively correlated with the course of HF (r =0. 703, P＜0.01 ), while Epac2 protein was negatively correlated (r = - 0. 409, P＜0.05). Conclusions During the progression of HF, Epac1 expression tends to decrease firstly and increase afterwards,while Epac2 expression declines continually. Epac may be involved in the pathogenesis of HF.

Objective To evaluate the relationship between histone deacetylase 3 (HDAC3) expression and the mechanism underlying mitigation of remifentanil postconditioning-induced protection of diabetic cardiomyocytes.Methods H9c2 cells were cultured in DMEM/F12 culture medium supplemented with 10％ fetal bovine serum.The cells were seeded in 6-well plates (2 ml/well) at a density of 105 cells/ml.After the cells were cultured for 12 h,the cells were attached to the wall and cultured for 48 h in the normoglycemic (5.5 mmol/L) or hyperglycemic (25 mmol/L) DMEM culture medium.The cells were then randomly divided into 6 groups (n =18 each) using a random number table:control group (group CON),hypoxia/reoxygenation group (group H/R),remifentanil postconditioning group (group RPC),hyperglycemia group (group HG),hyperglycemia plus hypoxia/reoxygenation group (group HG-H/R),and hyperglycemia plus remifentanil postconditioning group (group HG-RPC).In H/R,RPC,HG-H/R and HG-RPC groups,the cells were exposed to 95％ N2-5％ CO2 in an incubator for 5 h after changing the culture medium for Tyrode solution.In H/R and HG-H/R groups,the culture medium was changed to the DMEM/F12 culture medium supplemented with 10％ fetal bovine serum and glucose at the corresponding concentration,and the cells were then incubated for 1 h.In RPC and HG-RPC groups,the cells were incubated in the DMEM culture medium containing remifentanil at the final concentration of 1 μmol/L,and the cells were then incubated for 1 h.At 1 h of reoxygenation,the cell viability was measured by CCK-8 assay,the cell apoptosis was detected by AnnexinV-FITC/PI flow cytometry,and the expression of HDAC3 and caspase-3 in cells was detected by Western blot.The apoptotic rate was calculated.Results Compared with group CON,the cell viability was significantly decreased,the cell apoptotic rate was significantly increased,and the expression of caspase-3 and HDAC3 was significantly up-regulated in group H/R (P＜ 0.05).Compared with group H/R,the cell viability was significantly increased,the apoptotic rate was significantly decreased,and the expression of caspase-3 and HDAC3 was significantly down-regulated in group RPC (P＜0.05).Compared with group HG,the cell viability was significantly decreased,the apoptotic rate was significantly increased,and the expression of cspase-3 and HDAC3 was significantly up-regulated in group HG-H/R (P＜0.05).There was no significant difference in the parameters mentioned above between group HG-RPC and group HG-H/R (P＞0.05).Conclusion The mechanism underlying mitigation of remifentanil postconditioning-induced protection of diabetic cardiomyocytes is associated with hyperglycemia-induced up-regulation of HDAC3 expression.

AIM:To investigate the effect of AG 490 on the expression of VEGF and HIF-1α, and the capacity of invasion in human erythroleukemia (HEL) cells.METHODS:The HEL cells were treated with AG490 at different con-centrations .The cell viability was detected by CCK-8 assay.The apoptosis was detected by Hoechst staining .The apoptosis and the cell cycle were analyzed by flow cytometry .The capacity of migration was evaluated by Transwell assay .The mRNA expression level of JAK2 was measured by RT-PCR.The protein levels of p-JAK2, VEGF and HIF-1αwere determined by Western blot.RESULTS:The HEL cell viabilities were 88%, 75%, 48%, 10%and 0.12%after treated with AG490 at 20, 40, 60, 80 and 100 μmol/L for 48 h, respectively.The results of Hoechst staining showed that brilliant blue cells in 80 μmol/L AG490 group was significantly increased compared with control group for 48 h.The apoptosis rate of 80μmol/L AG490 group was significantly increased compared with control group at 48 h after AG490 treatment.The number of membrane-permeating HEL cells in 20μmol/L AG490 group at 24 h after AG490 treatment was significantly lower than that in control group (P<0.05).The mRNA level of JAK2 decreased in a concentration-dependent manner after the HEL cells were treated with different concentrations of AG 490 for 48 h.The protein levels of p-JAK2, VEGF and HIF-1αwere lower in AG490 treatment groups than those in control group (P<0.05).CONCLUSION: AG490 inhibits the expression of VEGF and HIF-1αin HEL cells by inhibiting JAK2 pathway.

Objective To evaluate the effect of dexmedetomidine on the postoperative cognitive function in the elderly patients with fragile brain.Methods One hundred and twenty elderly patients with fragile brain,aged 65-85 yr,weighing 50-80 kg,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ,with preoperative Mini-Mental State Examination score≥ 20,scheduled for elective gastroenteric surgical procedures,were randomly assigned to one of 2 groups (n =60 each) using a random number table:control group (group C) and dexmedetomidine group (group D).In group D,dexmedetomidine was intravenously infused in a loading dose of 0.4 μg/kg over 10 min before anesthesia induction,followed by an infusion of 0.4 μg · kg-1 · h 1until 30 min before the end of surgery.While the equal volume of 0.9％ nomal saline was given instead of dexmedetomidine in group C.Postoperative delirium was assessed within 3 days after operation using Confusion Assessment Method.Postoperative cognitive dysfunction was assessed at 7 days after operation using Mini-Mental State Examination.Results Compared with group C,the incidence of postoperative delirium was significantly decreased within 3 days after operation (P＜ 0.05),and no significant change was found in postoperative cognitive dysfunction at 7 days after operation in group D (P＞0.05).Conclusion Dexmedetomidine can decrease the occurrence of postoperative delirium in the elderly patients with fragile brain.

Objective To conduct a comprehensive evaluation on safety of Honghua Injection through adopting the method of the evidence-based method;To provide reference for clinical reasonable application of Honghua Injection. Methods Computers were used to retrieve some Chinese databases, such as China Biology Medicine, China National Knowledge Infrastructure, Wangfang Database and VIP database. At the same time, other search methods were employed, up to July 2013, including all research types about Honghua Injection. The adverse reactions in the reports of published literature were analyzed by description and statistical analysis. Results Sixty-nine researches on Honghua Injection were included. The total cases of adverse drug reaction (ADR) were 1111, among which male cases were 568 (51%), and female cases were 543 (49%). Thirty-six (52%) papers described ADR of Honghua Injection in detail, and thirty-three (48%) papers just mentioned ADR or did not describe ADR in detail. Skin, skin accessories damage and pathological changes in circulatory system were main contents of ADR. In terms of original diseases, diseases of circulatory system play an important role. Solvent medium was largely in line with its product specification requirements. Most ADR appeared when the drug was used for the first time, from 5 minutes to 5 days. Conclusion The current published literature data show that severe ADR does not happen after the intervention of Honghua Injection.

Objective To study the value of CT combined with immune tests for the diagnosis of cerebral schistosomiasis.Methods The data of 24 patients with cerebral schistosomiasis were collected and analyzed retrospectively,and all the patients were examined with CT and serum IHA and ELISA and,in addition,18 patients had the data of cerebrospinal fluid(CSF)IHA and ELISA.Results Twenty-one patients were diagnosed through CT and immune tests,one patients was diagnosed by postoperative pathology,and other two patients were proved by diagnostic therapy with praziquantel.Conclusion CT combined with immune tests has an important value for the diagnosis of cerebral schistosomiasis,and serum immune tests are more simple and more practical than CSF immune tests.

Citri Fructus identification by fingerprint and chemometrics was investigated in this paper. Twenty-three Citri Fructus samples were collected which referred to two varieties as Cirtus wilsonii and C. medica recorded in Chinese Pharmacopoeia. HPLC chromatograms were obtained. The components were partly identified by reference substances, and then common pattern was established for chemometrics analysis. Similarity analysis, principal component analysis (PCA) , partial least squares-discriminant analysis (PLS-DA) and hierarchical cluster analysis heatmap were applied. The results indicated that C. wilsonii and C. medica could be ideally classified with common pattern contained twenty-five characteristic peaks. Besides, preliminary pattern recognition had verified the chemometrics analytical results. Absolute peak area (APA) was used for relevant quantitative analysis, results showed the differences between two varieties and it was valuable for further quality control as selection of characteristic components.
Chromatography, High Pressure Liquid ; Citrus ; chemistry ; classification ; Discriminant Analysis ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; classification ; Mass Spectrometry ; Principal Component Analysis