Objective To construct a recombinant lentiviral vector for human thymosin ?4 (TMSB4) and to test the mRNA and protein expression of TMSB4 in 293T cells after being infected by shRNA lentivirus. Methods We designed a specific sequence of small hair RNA targeting TMSB4 gene; the complementary DNA containing both sense and antisense oligo DNA of the targeting sequence was cloned into the pGCSIL-GFP vector to construct a lentiviral vector. The vector was converted into the competent DH5a coli,which confirmed by PCR and sequencing. Then the viral vector and the packed systemic vector were cotransfected 293T cells to get the lentivirus. The virus titer was determined. Afterwards,in the 293T cells infected with the lentivirus,the expression of TMSB4 was detected by real time PCR and immunocytochemistry. Similarly,a primary fibroblast was also infected with the lentivirus. Results Compared with negative control cells,the mRNA and protein levels of TMSB4 in 293T cells infected with the lentivirus were reduced by 44% (0.56?0.11 vs 1.00?0.06,F=89.673,P

OBJECTIVE:To study the mechanism of improvement effects of exenatide on mitochondrial function of H9c2 cells under hypoxia/reoxygenation (H/R) condition. METHODS:H9c2 cells were cultured in vitro and were divided into blank control group,drug control group(exenatide 200 nmol/L),model group(H/R),pretreatment group(exenatide 200 nmol/L+H/R),gluca-gon-like peptide-1 receptor (GLP-1R) inhibitor [Exendin-(9-39) 100 nmol/L+exenatide 200 nmol/L+H/R], cAMP inhibitor (Rp-cAMPS 1 μmol/L+exenatide 200 nmol/L+H/R)group and PKA inhibitor(H-895 μmol/L+exenatide 200 nmol/L+H/R)group. Except for first 2 groups,H/R model was established in other groups,and they were given exenatide 30 min before modeling and relevant inhibitor 10 min before giving exenatide. Morphology of mitochondria was observed by TEM,and mitochondrial calcium (Ca2+m)and the mitochondrial membrane potential(ΔΨm)were determined by flow cytometry. Cellular ATP content was measured by microplate reader. RESULTS:Compared with blank control group, mitochondrial cristae swelling was enhanced in model group,while density decreased,showing vacuolization;Ca2+m level increased while ΔΨm and ATP decreased (P<0.05). Com-pared with model group,mitochondrial cristae swelling relieved in pretreatment group,while density increased,showing vacuoliza-tion relieved;Ca2+m level decreased,while ΔΨm and ATP increased(P<0.05). Compared with pretreatment group,the levels of Ca2+m increased in 3 kinds of inhibitors group,while ΔΨm and ATP decreased(P<0.05). CONCLUSIONS:Exenatide attenuates H9c2 cell mitochondria Ca2+m accumulation,increases ΔΨm and ATP production. Which indicate its mechanism may be associated with activating GLP-1R/cAMP/PKA pathway.

Objective To explore extracellular signal-regulated kinase ( ERK1/2 ) expression in the role of curcumin inhibited staurosporine (STS)-mediated neurons toxic injury through added PD098059, and to clarify ERK1/2 mediated inhibitory role of curcumin on STS-induced neurons toxic injury.Methods The neurons toxic injury model of primary cultured hippocampal neurons was established by STS.The experiment was divided into six groups:normal control group, STS model group, PD098059 +STS model group, curcumin +STS pretreatment group, curcumin+PD098059+STS treatment group and curcumin treatment group.The cell viability were determined by MTT method, lactate dehydrogenase (LDH) release rate, cell toxicity were detected, nuclear shape were observed by DAPI staining, and ERK1/2 expression were detected by Western blot method.Results The cell viability of curcumin +STS pretreatment group was significantly higher than STS model group ( P <0.001 ); the cell viability had no significant difference between PD098059 +STS model group and curcumin +PD098059 +STS treatment group;compared with curcumin +STS model group , the cell viability of curcumin +PD098059 +STS treatment group was significantly decreased ( P<0.001 ).LDH results showed that the nerve cell toxicity of curcumin +STS pretreatment group was obviously less than STS model group (P<0.001).The cell nuclear shape showed typical apoptosis morphological characteristics in STS model group, and curcumin inhibited the effect of STS mediated-neuronal apoptosis.ERK1/2 protein expression in curcumin +STS pretreatment group significantly increased compared with STS model group ( P <0.001 ) .Conclusion Curcumin inhibited STS-mediated neurons toxicity injury by up-regulating ERK1/2 expression.After PD098059 blocking the nerve cells ERK1/2 synthesis, the inhibitory action of curcumin failed to implemented, which illustrated that ERK1/2 mediated curcumin to inhibit STS-induced neuronal toxic injury.

Objective To analyze the usual death causes of cardiovascular disease and the differences in gender and age .Methods By adopting the retrospective study method ,the clinical data of death cases in cardiovascular disease were collected and analyze on the situation suffering from cardiovascular disease ,direct death causes ,gender and age difference .Results (1) among 181 cases of cardiovascular disease death ,coronary heart disease(115/181 ,64% ) and hypertension(96/181 ,53% ) were the most common dis-ease ,lung infection(104/181 ,57% ) was the most common complication ;(2)There was no significant difference in the situation suf-fering from basic diseases between male and female(P>0 .05);(3)The basic diseases in cardiovascular death cases aged over 60 years old were dominated by coronary heart disease and hypertension ;the proportion of complicating pulmonary infection was grad-ually increased with age increase ;(4) in the direct death causes ,the top 3 places were sudden cardiac death (44/181 ,24 .3% ) ,multi-ple organ dysfunction syndrome(24/181 ,13 .3% ) and cardiogenic shock(24/181 ,13 .3% ) .Conclusion Strengthening the manage-ment of diagnosis and treatment on elderly patients with coronary heart disease ,hypertension ,especially those complicating diabe-tes ,strengthening the treatment intervention of lung infection in cardiovascular disease population and conducting the emphasis pro-tection on the target organ function may reduce the mortality of cardiovascular inpatients .

ABSTRACT:Objective To a rare case of double primary cancer (hepatocellular carcinoma with gallbladder)to guide clinical application.Methods and Results We reported one case of primary hepatocellular carcinoma and gallbladder.A male patient,54 years old,had main complaints of intermittent right upper quadrant pain for 4 days. The abdominal CT of the local county hospital showed gallstones,gallbladder with liver infiltration.And then he went to the First Affiliated Hospital of Xi’an Jiaotong University for further treatment.Laboratory examination revealed:HBsAg(+),HBcAb(+),alpha-fetoprotein (AFP)> 60 500 ng/mL,carcinoembryonic antigen (CEA) 5.25 ng/mL.Abdominal CT showed hilar slightly stronger light echo groups:liver cancer or gallbladder cancer? Hepatic artery and portal vein CT imaging (CTA+CTV)examination showed the malignant tumor shadow in the inside of the left hepatic lobe huge,uneven thickening of the gallbladder wall,suspected liver disease with gallbladder infringement or gallbladder disease with liver infringement. With the preoperative preliminary consideration of primary liver cancer with infiltration of the gallbladder,we chose the operation as the resection of segment Ⅳ b and Ⅴ of the liver,cholecystectomy and T tube drainage.Pathological examination postoperative showed the bulky liver carcinoma grade Ⅲ and the poorly differentiated adenocarcinoma of gallbladder.A month later,abdominal CT showed the tumor spread intrahepatic,prompting the poor prognosis.Conclusion The two which are not continuous,which is the standard of double primary cancer,are not suitable for all double primary cancers.This case provides useful experience for future similar diagnosis and treatment of disease,and also helps us with timely and accurate identification of “metastatic”or “primary”,which is the key point for clinicians to give patients an effective treatment.

Objective To study the effect of neuroepithelial cell transforming gene 1 (Net1) on the cellular radiosensitivity and underlying mechanism.Methods Real-time quantitative PCR was used to measure the variations in Net1 expression level upon irradiation.Radiosensitivity was analyzed by colonyforming assay after Net1-siRNAs.Net1-associated proteins were identified by co-immunoprecipitation.Results The Net1 mRNA level in the cells was increased significantly (t =-10.52,P ＜ 0.05) after irradiation.Compared to the control group,siRNA-mediated silencing of Net1 enhanced cell radiosensitivity (t =15.31,11.65,P ＜0.05).Net1 was found to interact with Ku70,Ku80 and DNA-PKcs under either normal conditions or after irradiation.Conclusions Net1 could protect cells from irradiation by interaction with DNA repair proteins in non-homologous end joining pathway.

ObjectiveTo investigate the clinical efficacy and safety of Tetramethylpyrazine and Buthus martensi on children with cerebral daminfication syndrome.Methods39 cases with cerebral daminfication syndrome were randomly divided into the study group (18 cases, treated with Tetramethylpyrazine and Buthus martensi) and control group (21 cases, treated only with general nerve nutrition therapy). The curative effect of two groups was evaluated.ResultsAfter treatment, in the study group, 11 cases were cured, 4 cases more effective, 1 case efficacy, 2 cases inefficacy; in the control group, the numbers were 6 cases, 9 cases, 3 cases and 3 cases respectively. There was a significant difference between two groups (P<0.05). There was not side effect in all cases.ConclusionTetramethylpyrazine and Buthus martensi is safe and effective to treat children with cerebral daminfication syndrome.

For the purpose of exploring the teaching reform model and method, also promoting the quality of talents in acupuncture and tuina field, the research-based teaching model is applied into the course of Theory of Meridians and Acupoints. This method includes two parts of teaching and learning. For teachers, they bring modern research focus and trend into teaching through questionnaire survey among students, aiming to integrate the education inside and outside class. For students, they are guided to resolve the opening, enlightening and scalable issues through consulting abundant resources of literature and database to achieve autonomous participation in the course of learning. By this teaching method, it is expected to train the student's ability of expanding their thinking, as well as discovering, analyzing and solving the problem.
Acupuncture ; education ; methods ; Acupuncture Points ; Acupuncture Therapy ; methods ; psychology ; Humans ; Learning ; Meridians ; Musculoskeletal Manipulations ; education ; methods ; psychology ; Students, Medical ; psychology

Objective To study the rationality and possibility of 64 slice spiral CT in the examination of the temporal bone at low dose.Methods The same CT technique and temporal bone mode as those for clinical CT were used,two cranium specimens(four ears)were scanned with Somatom Sensation 64-slice spiral CT at different mA(380,300,200,160,120,80),and muhi-planar reformation was performed.The CT dose index at different mA groups were measured by 10 cm pencil ionization chamber and head dose phantom.Four anatomic structures on axial images(subarcuate fossa,tendon of tensor tympani, facial recess,etc),four anatomic structures on coronal images(scute,crista transversa,fenestra cochleae, etc)and six anatomic structures on double oblique images(malleus,incus,stirrup bone,upper bony semicircular,etc)were chosen to evaluate and grade the reformation images among different mA groups,and to determine the minimum mA value.Ten ears of five patients were used to test the validity of the minimum mA value.Results CT radiation dose was significantly reduced from(47.8?2.7)to(20.1?2.0)raCy (P

The contents about the central action mechanisms of needling Zusanli (ST 36) were sorted by summarizing the relevant literatures published in the past 10 years, and it was found that: by comparing acupuncture at Zusanli (ST 36), other acupoints or sham points, most studies showed that Zusanli (ST 36) had relatively specific characteristics in the brain function response; studies on the central action mechanisms of Zusanli (ST 36) were mainly focused on sequelae of cerebral apoplexy, pain, gastrointestinal diseases, weight loss and drug addiction withdrawal and so on; acupuncture manipulations, stimulation methods, individual differences, stimulation quantity, and stimulation duration, etc. could also induce different brain function responses; acupuncture at Zusanli (ST 36) had an after-working effect, also called long-term effect, but with poor repeatability. So, it was suggested that the future studies should focus on the factors that affect the clinical efficacies and experimental results, including the compatibility effects of the acupoints, reproducibility of research results, sample size, acupuncture methods, long-term effects and disease entities.