Objective To explore the over-expression of the CCAAT enhancer binding proteinα(C/EBPα)in hepatic stellate cells (HSC-T6) which activated by protease-activated receptor 1(PAR1) ,to explore its possible relationship with cell proliferation ,cell activation and collagen secretion of HSC-T6 cell .Methods HSC-T6 cells were divided into 4 groups ,control group ,SFLLR group;vehicle plasmid+SFLLR group ,C/EBPαplasmid+SFLLR group .The eukaryotic vector harboring the full length of C/EBPαcD-NA was transfected into PAR1 activated HSC-T6 cell ,then Western blot was applied to detect the expression of α-smooth muscle actin(α-SMA) and collagen I and evaluate the effect of transfection on proliferation of HSC-T6 by MTT .Results The expression of α-SMA and collagen I in C/EBPαplasmid+SFLLR group were dramatically decreased compared with other 3 groups ,as well as proliferation after 48 h(P<0 .05) .Conclusion Over-expression of C/EBPαgene by transfection had inhibitory influence on prolif-eration ,activation and collagen secretion of HSC-T6 which were activated by PAR1 .

This article expounds on the necessity of the introduction of comparative imaging in bone and joint imaging teaching according to the bone and joint imaging teaching situation and modern medical radiographic features.Meanwhile,it discusses the application of comparative imaging in bone and joint imaging teaching from aspects of teaching method,teaching effect evaluation and prospects of the comparative imaging.

Objective To investigate the value of Tei index for estimating right ventricular global function in patients with dissymmetry hypertrophic cardiomyopathy(including obstructive and nonobstructive).Methods Pulsed wave Doppler of transtricuspid flow and transpulmonic valve flow were performed in hypertrophic obstructive cardiomyopathy(HOCM,n=28),hypertrophic nonobstructive cardiomyopathy(HNCM,n=34), and healthy control subjects(n=30) at apical four-chamber view and short-axis view of aortic root,and Tei index was calculated.Results~①Accelerating contraction time/right ventricular ejection time(ACT/RVET),isovolumetric contraction time(ICT),isovolumetric relaxation time(IRT),isovolumetric contraction time+isovolumetric relaxation time(ICT+IRT),right ventricular ejection time(RVET),ACT/RVET and Tei index[(ICT+IRT)/RVET] showed no significant changes between the groups of HOCM and HNCM.②Tei index was distinctly different between patients with HCM and control subjects(P

Background and purpose:Radiotherapy (RT) is one of the most important therapeutic tools for esophageal cancer. Because tumors are heterogeneous, including for18F-FDG uptake and, most likely, for radioresistance, selective boosting of high FDG uptake zones within the tumor has been suggested. Therefore, it is critical to know whether the location of these high FDG uptake patterns within the tumor remains stable during RT.Methods:Twenty-two patients with esophageal squamous cell carcinoma treated with concurrent chemo-radiation underwent repeated18F-FDG PET-CT scans before RT and after 20 fractions of RT. On all scans, the high and low FDG uptake regions were auto-delineated using several standard uptake value (SUV) thresholds, varying from 40% to 70% of SUVmax on the pretreatment scan [gross tumor volume (GTV)40%pre, GTV50%pre, GTV60%pre, GTV70%pre] and from 70% to 90% of SUVmax on the dur-treatment scan (GTV70%dur, GTV80%dur, GTV90%dur) and ifxed thresholds of 2.5 and 5 (GTV2.5pre, GTV5pre). The volumes and overlap fractions (OF) of these delineations were calculated to demonstrate the stability of the high FDG uptake regions during RT.Results:The high uptake regions within the tumor during RT largely corresponded (OF>70%) with the 50% SUVmax high FDG uptake area (GTV50%pre) of the pretreatment scan. The hotspot within the residual area (GTV90%dur) was completely within the GTV and pre-radiotherapy high uptake regions (OF=100%). Although the location of the high FDG uptake patterns within the tumor during RT remained stable, the delineated volumes varied markedly.Conclusion:The location of the high FDG uptake areas within the tumor remained stable during RT. This knowledge may enable selective boosting of high FDG uptake areas within the tumor.

Objective To explore the effect of Xiaoyao pill combined with low-dose corticosteroid on serum estrogens levels in the treatment of patients with menopause syndrome and its clinical efficacy.Methods 96 patients with menopause syndrome from April 2013 to February 2015 in department of obstetrics and gynecology of Lishui second people’s hospital were selected and divided into 2 groups according to a random number table, the control group (48 cases) were treated with small dose of hormone therapy, the treatment group (48 cases) were treated with Xiaoyao pill combined with small dose of hormone therapy.The serum estrogens levels, Kupperman score, endometrial thickness and clinical efficacy were compared. Results The total effective rate of control group(79.17%) was lower than that of treatment group(93.75%)(P<0.05).Compared with control group post-treatment, the serum follicle stimulating hormone (FSH) and luteinizing hormone (LH) levels were lower, estradiol (E2) level was higher, the Kupperman score was lower, the endometrial thickness was higher than those of control group, there were significant differences ( P <0.05 ) . Conclusion Xiaoyao pill combined with small dose of hormone could improve serum E2, FSH and LH levels in the treatment of patients with menopause syndrome with high security.

Objective To investigate the effect of different doses of protamine neutralizing heparin on perioperation of on-pump coronary artery bypass graftting (CABG).Methods A total of 180 on-pump CABG patients hospitalized from January 2015 to November 2016 were randomly divided into three groups,the protamine group l,protamine group 2 and protamine group 3,60 patients in each group.Heparin (3 mg/kg) was used before extracorporeal circulation.After intracardiac operation was over,protamine was used to neutralize the heparin to adjust the activated clotting time (ACT) in protamine group 1,which was 10％ higher than that of intubation.Meanwhile,protamine group 2 was neutralized to equal to the ACT before intubation,and protamine group 3 was 10％ lower than that before the intubation.The differences of intraoperative and postoperative parameters were compared between the three groups.Results No death was found in the three groups during hospitalization.Comparing with protamine group 1 and protamine group 2,the time of operation,the ACT before the leaving operation room,the ACT of the first hour after returning to ICU,the amount of bleeding during operation,the time of closing and the amount of red blood for transfusion were decreased in protamine group 3 (P ＞ 0.05).The total amount of protamine for neutralizing and the ratio of protamine and heparin were significantly increased in protamine group 3 (P ＜ 0.05).The heart dysfunction after operation,perioperative myocardial infarction,pulmonary edema,pulmonary infection,renal dysfunction,poor wound healing,neurological complications,and time of in hospital stay showed no significant differences between three groups (P＞0.05).Conclusion ACT below 10％ of preoperation is safe,after neutralization of heparin by protamine,which can obviously reduce the bleeding,the time of sternal closure and the amount of red blood cell transfusion,showing a positive clinical significance.

This paper introduced the community service of industrial injury insurance in 3 years in a community in Zhuzhou, Hunan, and the problems and difficulties of the community service. 4 kinds of advice were suggested to improve the community service.

BACKGROUND:As current studies on isolation, culture andcryopreservation of human amniotic epithelial cells (hAECs) are relatively scattered, it is difficult to form a comprehensive and effective solution to meet the clinical needs of stem cells for transplantation in future.OBJECTIVE:To establish the technology of isolation, culture and cryopreservation of hAECs, and to study the biological characteristics of hAECs.METHODS:Orthogonal method was used to study the effects of different factors on the separation, culture and cryopreservation, and range method was adopted to analyze the data to optimize the separation, culture and cryopreservation. We performed cell primary and passage cultures, morphology observed by microscope, drawn cell growth curve and flow cytometry assay, immunofluorescence staining, hepatocyte like cell differentiation to study the biological characteristics of hAECs.RESULTS AND CONCLUSION:(1) The optimal hAECs separation conditions were as follows:trypsin digestions were conducted at a concentration of 0.25%, four times, once for 20 minutes digestion; optimal conditions of culture were 4×108/L cell seeding density, 10 μg/L epidermal growth factor, 5% serum; optimal conditions of cryopreservation were 1×1010/L cell cryopreservation density, 10% dimethyl sulfoxide, 80% serum. (2) The primary cells were adhered to the wall in 2-3 days, exhibiting irregular polygon, paving stone-like growth. Cell adherence and growth rate were accelerated after subculture, and the growth and proliferation ability of passage 2 cells were not significantly decreased after cryopreservation and resuscitation. (3) Immunofluorescence staining showed that the primary cells strongly expressed SSEA-4 and CK19, but did not express Vimentin, CD45 and HLA-DR. The immunophenotype statistics of the primary and passage 4 cells showed the epithelial mesenchymal transition of hAECs in culture process. (4) Immunofluorescence staining showed that the liver cell marker expression of ALB, CK18 was significantly increased after hAECs were induced to differentiate into hepatocyte-like cells. Glycogen staining revealed glycogen synthesis in hAECs after 3 weeks of induction. To conclude, hAECs are easy to obtain and have strong proliferation ability in vitro, and express surface markers for undifferentiated embryonic stem cells.

Objective To construct the genetic recombinant adenovirus vector carrying the human growth and differentiation fac-tor-5(GDF-5) gene by using AdEasy-1 adenovirus vector system and to amplify and prepare the recombinant adenovirus in HEK 293 cells .Methods Human GDF-5 gene obtained by PCR was inserted into plasmid pMD19-T and the 1 .7 kb GDF-5 cDNA sequence was cloned into the adenoviral shuttle plasmid pShuttle-cytomegalovirus(CMV) of the AdEasy-1 system .It was identified by DNA sequencing and a digestion with Hind Ⅲ restriction enzyme .The resultant pShuttle-CMV-GDF-5 was used to generate the adenovi-ral GDF-5 vector through homologous recombination with the adenoviral backbone plasmid ,pAdEasy-1 in BJ5183 bacterial cells .It was selected by kanamycin and identified by a digestion with Hind Ⅲ restriction enzyme and amplified in XL10-Gold competent bac-teria .The DNA of recombinant adenovirus vector was finally linearized by Pac Ⅰ and the adenoviral recombinants were used to pro-duce adenoviruses in HEK293 packaging cells ,resulting in an Ad-GDF-5 identified by Western blot .The virus titer was assayed by TCID50 .Results GDF-5 cDNA sequence obtained by PCR was 1 .7 kb .Gene sequencing results indicated that the sequence was i-dentical with the one in GENBANK .Cloned sequence 1 .7 kb(GDF-5) was obtained by a digestion with Hind Ⅲ restriction enzyme after GDF-5 cDNA segment was cloned into pShuttle-CMV and AdEasy-1 .Western blot showed that two bands migrating at ap-proximately 15 and 55 kDa were clearly observed in PVDF membrane .These data confirmed that HEK293 cells expressed a large number of mature GDF-5 protein after infected with Ad-GDF-5 .Our research results demonstrated that recombinant adenovirus vector GDF-5 was successfully constructed .The virus titer was 5 .6 × 109 PFU/mL .Conclusion Recombinant adenovirus vector carrying the human GDF-5 gene is successfully constructed by using the AdEasy-1 adenovirus vector system .Moreover ,the Ad-GDF-5 with high titer is prepared .These provide the basis for further study of the biological function of GDF-5 and the gene thera-py of its related diseases .

OBJECTIVE:To investigate clinical efficacy and safety of meglumine adenosine cyclophosphate in the treatment of acute myocardium infarction. METHODS:A total of 80 patients with acute myocardium infarction in our hospital during May 2015-Jan. 2016 were selected and divided into control group and observation group according to random number table,40 cases in each group. Control group was given conventional treatment. Observation group was additionally given Meglumine adenosine cyclo-phosphate injection 120 mg added into 5% Glucose injection 250 mL,ivgtt,qd,on the basis of control group. Both groups re-ceived treatment for 7 d. Clinical efficacies as well as the levels of serum hs-CRP and NT-proBNP before and after treatment were observed in 2 groups,and the occurrence of ADR was compared between 2 groups. RESULTS:Total response rate of observation group(92.50%)was significantly higher than that of control group(75.00%),with statistical significance(P<0.05). Before treat-ment,there was no statistical significance in the serum levels of hs-CRP or NT-proBNP between 2 groups(P>0.05). After treat-ment,the serum levels of hs-CRP and NT-proBNP in 2 groups were decreased significantly,and the observation group was signifi-cantly lower than the control group,with statistical significance(P<0.05). There was no statistical significance in the incidence of ADR between 2 groups(P>0.05). CONCLUSIONS:Meglumine adenosine cyclophosphate shows significant therapeutic efficacy for acute myocardium infarction,reduces serum levels of hs-CRP and NT-proBNP significantly with good safety.