In order to detect the expression of RhoA in placenta from normal pregnancy and preeclampsia and evaluate the role of RhoA in preeclampsia, the expression of RhoA in placenta collected from 40 preeclampsia patients and 20 normotensive controls was determined by immunohistochemistry and RT-PCR. RhoA was found in syncytiotrophoblasts and cytotrophoblasts. The mean density of RhoA protein in placental tissues of mild and severe preeclampsia groups was significantly higher than that in normal pregnancy. The expression level of RhoA mRNA in mild and severe preeclampsia groups was significantly higher than that of normal pregnancy. Increased expression of RhoA in placental tissues might play an important role in the pathogenesis of preeclampsia.

Objective In order to study the biological significance of HBV/C gene variant in vitro.Methods Retroviral vector pXT1 was used to construct the HBV/C gene expression vector and the recombinant plasmid pXT1-HBV/C was used to transfect immortalized human peripheral blood B cell lines to express HBcAg steadily in the host cells.Results plasmid pXT1-HBV/C was detected positive by PCR as well as enzyme digestion with Bgl Ⅱ and Xho Ⅰ.Meanwhile.transfected cells was detected to contain HBV/C gene by PCR and HBcAg was expressed in 47.4%of the ceils by means of flow cytometry.Conclusion Retroviral expression vector with HBV/C gene can transfeet into eucaryotic cells effectively and express the goal gene steadily.The recombinant cells may be used in the systematic studies on the biological significance of HBV/C gene variant.

Objective To study the biological significance of the common mutant preC/C gene in clinical HBV in China. Methods Site directed mutagenesis based on the unique enzyme site elimination was used to construct eukaryocyte expression vector with mutant HBV/C gene(V60、G87、L97). Expression vectors with wild and mutant preC/C gene were transferred into HepG2 cell. Culture supernatant was detected by ELISA for HBeAg. Results Result of DNA sequencing showed that the constructed mutant HBV preC/C gene had only one specific site variation compared with the wild type sequence. Goal DNA fragment was detected positive in the HepG2 cells transferred with wild and mutant preC/C gene. A value of HBeAg in the supernatant of the cells harboring L97 variant was higher than that of the wild and other variant strains( P

To study the characteristics of EBV transformed human peripheral blood B cell lines from hepatitis B patients and to provide a basis for further studies on the cellular immune function of hepatitis B patients.High titer of EBV was produced by B95 8 cell by induction with sodium n butyrate,and peripheral mononuclear cells from two different groups of hepatitis B patients were harvested and infected with EBV. The results showed that 4～8 weeks after infection 19 EBV transformed B cell lines from hepatitis B patients were established. It was found that colony formation of cells from A group appeared 2～3 weeks later than B group. Cells grew healthily and had good activity after 4 weeks of freezing.CD19 and CD20 were detected in 88 34% and 37 48% of the immortalized B cell membrane respectively. HBV DNA could not be detected in a11 19 immortalized B cell lines.It suggested that the time needed for the establishment of B cell lines was related with the immune state of the patients.HBV DNA could not exist persistently, and it would disappear finally in the immortalized B cell lines.

In the light of mathematical maximum likily hood method, the conditioned probability of clinical manifestations in TCM syndrome of asthenic heart qi - and blood and control group were calculated to establish a " List of Quantitative Diagnostic Index of TCM Manifestations Differentiation for Qi and Blood in Heart Disease". Based on the scoring of the above list, the rate of coincidence of retro - spective test of the three signs were 92. 70%, 93. 20%, 94. 98% respectively, and that of prospective test were 84. 61%, 81.82%, 87. 50% respectively. Authors' address: Institute of diagnostics, Hunan College of TCM, Changsha, Hunan 410027

Objective To investigate clinical outcomes and complications of isolated polyethylene liner exchange for revision total hip arthroplasty.Methods From April 1995 to December 2007,80 patients (93 hips) underwent revision total hip arthroplasty during which only polyethylene liner was exchanged with reservation of acetabular cup.There were 41 males and 39 females,aged from 27 to 82 years (average,53.3 years).The duration from the primary THA to the revision surgery ranged from 0.3 to 18.4 years (average,10.9 years).The reasons for liner exchange included:polyethylene wear with osteolysis (78 hips),polyethylene wear without osteolysis (5 hips),polyethylene wear with stem loosening (4 hips),recurrent dislocation (3hips),infection (1 hip),periprosthetic fracture (1 hip) and liner dislodgement (1 hip).Forty-seven liners were fixed into the old cup using cement,and 46 were fixed with the original locking mechanism.Sixty cross-linked polyethylene liners and 33 conventional polyethylene liners were used.Results All patients were followed up for 5 to 15 years (average,7 years).The average Harris hip score improved from preoperative 86.0±16.9 to 89.4±11.6 at final follow-up.Complications included dislocation (10 hips),infection (2 hips),periprosthetic fracture (1 hip) and liner dislodgement (1 hip).Ten hips underwent rerevision due to different reasons:cup exchange (5 hips),conventional polyethylene wear (2 hips),infection (2 hips) and liner dislodgement (1 hip).Using component loosening as the end point,the 10-year survival rate was 100％ in the cement fixation group and 84.8％ in the original locking group.Using rerevision as the end point,the 10-year survival rate was 90.4％ in the cement fixation group and 65.0％ in the original locking group.Conclusion Liner exchange either with cement or original locking mechanism is a safe and successful method.Highly cross-linked polyethylene has a higher wear resistance,which can reduce incidence of osteolysis and improve survival rate of prosthesis.

This study examined the association of expression of vascular endothelial growth factor (VEGF), a promoter of angiogenesis, with endothelium dysfunction in preeclampsia. The level of VEGF protein and mRNA in the placenta and peripheral blood samples of 30 preeclampsia patients and 30 normotensive pregnant women was measured by immunohistochemistry, real-time reverse transcriptase-polymerase chain reaction (RT-PCR) and enzyme-linked immunosorbent assay (ELISA), respectively. VEGF expression in the human umbilical vein endothelial cells (HUVECs) was blocked by small interfering RNAs (siRNAs). The monolayer barrier function of HUVECs was determined by measuring the fluorescence intensity of BSA that crossed the HUVEC monolayers. The cell proliferation and cell-secreted nitric oxide (NO) level were detected by MTT method and nitrate reductase assay, respectively. The results showed that VEGF was expressed in the syncytiotrophoblasts and endothelial cells of vessels and capillaries in the placenta tissue. The serum level of VEGF in the preeclampsia patients was significantly decreased as compared with that in normal pregnant subjects, although VEGF mRNA expression in the placenta tissue of preeclampsia patients remained still high. Moreover, VEGF deficit could lead to endothelium cell dysfunction, and the administration of VEGF could protect endothelium cells from injury. It was concluded that lack of VEGF contributes to endothelium dysfunction, which may lead to the occurrence and development of preeclampsia.

In order to prevent the teaching reform of organ system curriculum from fall of teaching quality, the teaching effect of organ system curriculum and subject curriculum system in teaching the basic knowledge of pathophysiology was compared. In organ system curriculum teaching, there was no difference between the grades of students in Batch 2011 and Batch 2012 who conformed to the teaching principle of "gradual improvement" and those of the students taught with subject curriculum system. On the contrary, the students of Batch 2013 and Batch 2014 with insufficient curriculum content integration had a decreasing trend or a significant reduction in the teaching effect of organ system curriculum compared with that of subject curriculum system. After the supplementary for curriculum knowledge was made, the teaching effect of the organ system was significantly improved, which was better than that of the subject curriculum system. In conclusion, we have summarized and reflected on the effectiveness of the teaching reform of organ system curriculum, once again proved that basic medical teaching must also follow the inherent law of medical education, which is the teaching principle of "gradual improvement."

OBJECTIVETo investigate the role of the 25 kD hepatitis B e antigen (HBeAg) precursor that only exist inside hepatocytes and study its effect on the pathopoiesis of hepatitis B and QIAGEN expression and purification system.

METHODSHepatitis B virus (HBV) preC/C gene for the 25 kD HBeAg precursor was cloned into the expression vector pQE30 and the 25 kD HBeAg precursor was expressed in Escherichia coli (E. coli) and purified. Its antigenicity for 21 kD mature HBeAg was tested by western blot analysis.

RESULTSCloned fragments in the expression vector were sequenced and verified to be homogeneous with that of HBV (ayw subtype). Expression of the HBeAg precursor in E. coli under the transcriptional regulation of T5 promoter yielded a soluble cytosolic protein with an apparent molecular mass of 25 kD. Recombinant HBeAg precursor exhibited identical potencies with 21 kD mature HBeAg that reacted with anti-HBeAg antibodies. The purification rate of the expressed HBeAg precursor was up to 89.6% and the yield of purified HBeAg precursor from this procedure was 2.4 mg/L.

CONCLUSION25 kD HBeAg precursor exhibited biological activity and might play an important role in pathopoiesis of hepatitis B.

Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Escherichia coli ; genetics ; Gene Expression ; Hepatitis B e Antigens ; genetics ; isolation & purification ; metabolism ; Plasmids ; genetics ; Protein Precursors ; genetics ; isolation & purification ; metabolism ; Recombinant Proteins ; isolation & purification ; metabolism

OBJECTIVETo analyze the predictive factors of the therapeutic effects of interferons (IFNs) and entecavir (ETV) treatments for 48 weeks in patients with chronic hepatitis B (CHB) positive for HBeAg.

METHODSThis retrospective analysis compared the treatment efficacy of IFNs and ETV in 129 CHB patients positive for HBeAg. Twenty-seven of the patients were treated with PEG-IFNα-2a (180 µg once a week, PEG-IFN group), 51 patients with conventional IFNα (5 MIU three times a week, IFN group), and 51 with ETV (0.5 mg once daily, ETV group) for 48 weeks.

RESULTSAfter completion of the treatment cycles, the patients in ETV group showed a significantly higher HBV DNA undetectable rate and a significantly lower HBeAg seroconversion rate than those in PEG-IFN and IFN groups (P<0.05); HBeAg seroconversion rates were similar between PEG-IFN group and IFN group (Χ(2)=0.709, P=0.400). In PEG-IFN and ETV groups, HBeAg seroconversion rates were not associated with age, gender, baseline HBeAg, baseline HBV DNA and baseline ALT. In IFN group, HBeAg seroconversion rates were associated with baseline HBeAg (P=0.048) but not with age, gender, baseline HBV DNA and baseline ALT. In PEG-IFNα-2a group, ROC analysis showed that the sensitivity and specificity of HBeAg seroconversion at 48 weeks were 0.778 and 0.889, respectively, when the decline rate of HBeAg between baseline and week 24 exceeded 97.81%, with the corresponding positive and negative predictive values (PPV and NPV) of 0.778 and 0.889, respectively; the sensitivity and specificity of HBeAg seroconversion at 48 weeks were 0.889 and 0.722, respectively, when the decline rate of HBeAg between week 12 and week 24 was over 42.75%, with the corresponding PPV and NPV of 0.615 and 0.929, respectively.

CONCLUSIONTreatments with PEG-IFNα-2a and conventional IFNα for 48 weeks can achieve a higher HBeAg seroconversion rate than ETV, but the latter produces a higher HBV DNA undetectable rate. For PEG-IFNα-2a treatment, the decline rate of HBeAg between baseline and week 24 over 97.81% is the best predicting factor for HBeAg seroconversion at week 48 in CHB patients positive for HBeAg.

Adult ; DNA, Viral ; blood ; Female ; Guanine ; analogs & derivatives ; therapeutic use ; Hepatitis B e Antigens ; blood ; Hepatitis B, Chronic ; blood ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; therapeutic use ; Retrospective Studies ; Young Adult