To establish a cell model in vitro that stable expressing HBV by integrating HBA1.3 DNA into cell chromosome. The HBV1.3 full-length DNA was obtained by digested pGEM-HBV1.3 plasmid with HindIII and then was linked with PU-21 vector digested by HindIII. This was resulted in generation of a recombined plasmid named PU21-HBV plasmid. The recombined plasmid was introduced into HepG2 cells by electroporation. The transfected cells were screened with G418. The insertion and expression of HBV were identified by X-gal staining, RT-PCR and Southern blot. The result of PU21-HBV plasmid sequence demonstrated that HBV1.3 DNA was linked correctly with PU-21 vector. A series of positive cell colonies were obtained with G418 screening followed transfecting PU21-HBV plasmid into HepG2 cells. The results of Southern blot and RT-PCR exhibit that HBV1.3 DNA had successfully integrated into the chromosomes of HepG2 cells and had functional HBV gene transcription. HBV1.3 DNA was inserted into HepG2 genome and could stable transcript HBV RNA. The stable HBV expression cell line was constructed successfully. There are LoxP sites in the trapping vector PU21. With the Cre enzyme, interesting genes could be excganged into the LoxP sites. Therefore, double stable expression of interesting gene and HBV cell lines could be generated. The cell lines will be useful for further research some target gene function on replication of HBV.

OBJECTIVE: To explore the relationship between the expression of EIIIA+ fibronectin in incised wound of rat's skin and injury time. METHODS: The wounding model was established by cutting the dorsal skin of 48 adult SD rats. The rats were sacrificed at the pre-set injury time as immediately, 0.5 h, 1 h, 2 h, 3 h, 4 h, 6 h, and 8 h. The skin samples were taken at the margin of wound. The expression of the EIIIA? fibronectin was detected by immunohistochemistry and Western blotting and the relationship be- tween its expression and injury time was observed. Results The expression of EIIIA+ fibronectin was not observed immediately. The basal cell of skin began to show positive expression 0.5 h after injury. With the extension of injury time, positive staining became stronger. The value of relative optical density was gradually increased with prolonged injury time by the Western blotting analysis. CONCLUSION The expression of EIIIA+ fibronectin could be used for estimation of injury time in the early stage of skin injury.
Animals ; Fibronectins/metabolism* ; Immunohistochemistry ; Proteins/metabolism* ; Rats ; Rats, Sprague-Dawley ; Skin/metabolism* ; Staining and Labeling ; Time Factors

In order to demonstrate PTB bind to HPRE,reverse transcription,PCR-mediated detection,were used.HepG2.2.15 cell line and HBs-HPRE transient expression cells were adopted to identify PTB function in HBV life cycle.The results showed that PTB could directly bind to HPRE RNA.Functional analysis indicated that PTB could inhibit the expression of HBs antigen and this inhibition was in a dose-dependent manner in HepG2.2.15 cells.Higher expression of HBs in cells transfected pcDNA3-HBs-HPRE comparing with pcDNA3-HBs,and this high expression could also be inhibited by PTB.The data demonstrated that PTB inhibits HBs expression by interacting with HPRE.

OBJECTIVETo observe the changes of the cytokines following stereotactic irradiation for hepatocarcinoma with cirrhosis in rabbits.

METHODSSixteen rabbits with liver cirrhosis and hepatocarcinoma (experimental group) were randomized into two equal groups to receive stereotactic irradiation at single dose of 20 and 30 Gy, respectively. Eight rabbits with hepatocarcinoma (control group) were divided into two equal groups and treated in identical manner. All the rabbits were killed 3 weeks after irradiation, and EV two-step method was used to observe the cytokine changes of proliferating cell nuclear antigen (PCNA) and glutathione S-transferase pi (GST-pi) after irradiation.

RESULTSAfter irradiation, PCNA and GST-pi expression showed significant difference in the adjacent liver tissue between the experimental and control rabbits with irradiation at 20 Gy (P=0.010), but not with the irradiation dose of 30 Gy (P=1.000). Irradiation at different doses resulted in significant difference in the cytokine expression in the experimental rabbits (P=0.010). In the liver tissue exposed to irradiation, different irradiation doses resulted in significant difference in PCNA and GST-pi protein expression (P=0.010).

CONCLUSIONSFor hepatocarcinoma with cirrhosis in rabbits, radiation at the single dose of 30 Gy produces better response than 20 Gy, and PCNA and GST-pi may serve as good indexes for evaluating the therapeutic effect.

Animals ; Glutathione S-Transferase pi ; biosynthesis ; Immunohistochemistry ; Liver Cirrhosis, Experimental ; metabolism ; radiotherapy ; Liver Neoplasms, Experimental ; metabolism ; radiotherapy ; Male ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Rabbits ; Radiotherapy Planning, Computer-Assisted ; methods ; Radiotherapy, Conformal ; Random Allocation

Objective: Through analyzing the development practice and restricting factors of social capital running medical institutes in ethnic minority areas, it explored the regulation path for social capital running medical institutes in ethnic minority areas. Methods: Through researching the policy practice, hold practice and operation practice, it selected 8 ethnical provinces as research areas and summarized the development practice of social capital running medical institutes in ethnic minority areas. Results: The current constraints of social capital running medical institutes in China’s ethnic areas mainly reflected in the constraints of the economic depth of poverty to restrict the medical treatment of the patients in the society, the lack of policy guarantee restricted the competitiveness of the social medical market and the agglomeration of social medical personnel. Conclusion: Social capital running medical institutes in ethnical areas should regulate the path of institutional construction, the regulatory path of the industry and the regulatory path of PPP (Public Private Partnership) path.

AIM:To investigate the role of autophagy in inhibition of human lung cancer PC 9 cell proliferation by ursolic acid(UA)as well as the underlying mechanism.METHODS:MTT assay and Trypan blue exclusion test were performed to analyze the effect of UA on the proliferation of PC 9 cells.The PC9 cells were treated with UA,and autophagy was observed under fluorescence microscope through acridine orange staining.The expression of autophagy-associated pro-teins LC3 and ATG5 in the PC9 cells were detected by Western blot.The effect of UA,3-methyladenine(3-MA)3-MA or their combination on the cell viability was measured by MTT assay.RESULTS:The viability of PC9 cells was significantly inhibited by UA(P<0.05 or P<0.01).The number of bright red fluorescence positive cells was significantly increased after treatment with UA.The protein expression of LC3-II and ATG5 was significantly up-regulated compared with control group(P<0.01).Furthermore,combination of UA and 3-MA resulted in a substantial decrease in cell viability compared with using UA alone(P<0.01).CONCLUSION: UA inhibits the proliferation and induces the autophagy of the PC 9 cells,in which autophagy plays a protective role.The inhibition of autophagy significantly promotes the death of the PC 9 cells induced by UA.

Objective To explore the self-face recognition and its relationship to empathy in patients with schizophrenia.Methods Sixty-two schizophrenic patients and fifty -four healthy subjects were assessed with the self-face recognition task (SFRT) and the interpersonal reactivity index-C (IRI-C).Results The SFRT reaction time in the patients group( (2188 ± 1138) ms) was significantly longer than that in the control group( ( 1152 ± 326) ms) (P ＜ 0.01 ) ;the accuracy in the patients group ( (80 ± 16) ％ ) was significantly lower than that in the control group ( (88 ± 6) ％ ) (P ＜ 0.01 ).The IRI-C total scores,the subscores in perspective taking,the subscores in fantasy and empathic concern of IRI-C were significantly lower in the patients group(respectively(44.82 ± 10.50),(8.98 ± 3.56),( 11.87 ± 4.38 ),( 14.73 ± 4.00) ) than those in the control group ( respectively (49.85 ± 10.28),( 10.78 ± 3.86),( 14.98 ± 6.12),( 17.39 ± 4.56) ) ; the subscore in personal distress of IRI-C in the patients group(9.37 ± 5.12) was significantly higher than those in the control group(6.52 ± 3.89) ( P＜ 0.01 ).There was significant positive correlation between the accuracy for self-face recognition in SFRT and the subscore in fantasy of IRI-C ( r =0.322,P ＜ 0.05 ),the reaction time of SFRT had significantly positive correlation with the subscore in personal distress.Conclusion Schizophren patients have general impairments of self-face recognition and empathic abilities,and the self-face recognition is related to the empathic abilities.

Objective To study the clinical characteristics of hepatitis B virus-related acute-on-chronic liver failure patients with familial aggregation.Methods 275 patients with hepatitis B virus - related acute-on-chronic liver failure were investigated.The patients were divided into familial aggregation and nonfamilial aggregation group basis on their epidemiological features.Clinical data and biochemical indicators between the two groups were analyzed statistically.Results 93 of 275 patients( 33.82％ ) case were family aggregation.There was no significant difference compared with chronic hepatitis B patients( 38.3％ ).The mean age of the two groups was 45.98 and 43.61 years old,respectively (P ＞ 0.05 ).The rates of liver cirrhosis in family aggregation group were significant higher than non - familial aggregation group( 73.91％ vs 58.24％,p ＜ 0.05 ).Serum total (TBil) and prothrombin activities ( PTA ) were no significant difference between the two groups,but ALT level in familial aggregation group was much higher (407.80U/L vs 256.45 U/L,P0.05 ).Conclusion Familial aggregation were not related to acute-on-chronic liver failure in chronic HBV hepatitis patients. But the rate of liver cirrhosis were higher in patients with familial aggregation.

OBJECTIVETo explore the association between XAGE-1b gene expression and the clinical characteristics of non-small cell lung cancer (NSCLC).

METHODSTumor tissue and adjacent normal lung tissue specimens were obtained surgically from 30 patients with resectable NSCLC, from which the total RNA was extracted for RT-PCR to amplify full-length XAGE-1b gene. The products of RT-PCR were identified by electrophoresis and sequencing. The expression of XAGE-1b gene and its association with the clinical characteristics of the patients were analyzed.

RESULTSIn the 30 tumor tissue specimens, the expression rate of XAGE-1b gene was 40%, but none of the normal lung tissues expressed this gene. The gene expression was not related to the patients' age, gender, tumor differentiation or clinical stages, but showed significant correlation to their pathological classification. The expression rate of XAGE-1b gene in adenocarcinoma was much higher than that in tumors of other pathological types (61.1% vs 8.3%, P=0.015). XAGE-1b gene expression tended to increase with the TNM stages, which, however, failed to find statistical data support (P>0.05).

CONCLUSIONSXAGE-1b gene is highly expressed in lung adenocarcinoma, and can be an ideal target for tumor immunotherapy.

Antigens, Neoplasm ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lung Neoplasms ; genetics ; pathology ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo investigate the therapeutic effect of recombinant human parathyroid hormone(1-34) [rhPTH(1-34)] on osteoporosis of ovariectomized rats.

METHODSThe model of osteoporosis was formed after 3 months of ovariectomy with 6-month age of 80 rats. Another 20 rats was control of sham operation. rhPTH(1-34) was subcutaneously injected once daily with 5, 10, 20, 40 micrograms/kg for 3 months. There were 10 rats in each group. The control of therapy included Salmon Calcitonin to 10 rats and Alendronate sodium to 10 rats. The bone weight of dry and ash, bone mineral density, bone biomechanical property, trabecular area, bone mineral deposition and serum alkaline phosphatase, Ca, P and urinary Pyridinoline/creatin (Pyd/Cr) were measured after the end of therapy.

RESULTSWhen administered to animals as a single subcutaneous injection once daily, rhPTH(1-34) increased obviously bone mass, bone biomechanical property and trabecular area, as well as bone deposition compared with the animals of control group. The bone architecture was ultimately improved by rhPTH(1-34) therapy.

CONCLUSIONSRats of ovariectomized-induced osteoporosis possess obvious effect of treatment with low dose of rhPTH(1-34) administered once daily.

Animals ; Female ; Osteoporosis ; drug therapy ; etiology ; Ovariectomy ; Rats ; Recombinant Proteins ; therapeutic use ; Teriparatide ; therapeutic use