A set of specific primers was synthesized according to DNA sequence of HBV found in China, the reverse transcriptase (RT) region in polymerase gene was amplified by PCR method from the serum of 3 patients with chronic HBV infection, and then the PCR products were subcloned into pGEM Teasy vectors. 13 clones were sequenced. Sequence comparison of the selected clones was made to look for the difference. After being compared, 13 sequences of RT were found different. Besides 2 clones with long sequence deletion, the different rate of RT coding nucleic acid sequences, RT and HBsAg amino acid sequences of the 11 clones is 5 1%, 4 9% and 7 5%, respectively. Many mutation types, including point substitution and deletion mutation, were found in this region. There is quasispecies population and defective HBV genome in patients with chronic HBV infection.

Background Researches have demonstrated that ocular hypertension induces the ischemia-reperfusion of retina and further leads to the degeneration of retinal ganglion cells,but its mechanism is beyond understanding.Objective The present study aims to observe the effects of static pressure on the morphology,proliferation activity and viability of cultured retinal microvascular endothelial cells (RMECs) and evaluate the expression of ET-1 and NO in these cells under variant static pressure.Methods RMECs were isolated from 30 healthy Wistar rats and cultured using explant culture method by Ⅷ factor antibody and PECAM-1 antibody.The static pressure of 1.33kPa,2.67kPa,5.33kPa and 10.67kPa was used in culture bottle respectively.The RMECs without static pressure were used as normal control group.The morphology of RMECs under the different static pressure was observed by inverted phase contrast microscopy,and the number of RMECs was counted using the counting plate.Cellular viability was studied by trypan blue staining.The changes of ET-1 and NO_2~-/NO_3~-,two metabolic products of NO,in the medium were detected by radioimmunoassay and Griess's nitrate reductase method.The expression of ET-1,eNOS and iNOS mRNA in RMECs was analyzed by semi-quantitative RT-PCR 24 hours after treatment of variant static pressure.Results Cultured RMECs sticked well at 24 hours and reached to confluence at 48 hours and showed the red fluorescence for Ⅷ factor antibody and PECAM-1 antibody.Enlargement of nuclei,extenders of cell bodies and suspension of RMECs in medium were observed.The number of RMECs was gradually increased.The cell viability was reduced with the raise of static pressure among these four groups(F=12.205,P＜0.01;F=11.180,P＜0.01).The static pressure increased the content of ET-1 released by RMECs in 2.67kPa,5.33kPa and 10.67kPa of static pressure groups,and concentrations of NO_2~-/NO_3~- in the medium showed a significant increase in 5.33kPa and 10.67kPa of static pressure groups compared with normal and 1.33kPa of static pressure groups(P＜0.01).The expressions of ET-1 mRNA,eNOS mRNA and iNOS mRNA were considerably enhanced in 5.33kPa and 10.67kPa of static pressure groups compared with normal control group(P＜0.01).Conclusion Raised static pressure causes the alteration of RMGCs structure and morphology.Static pressure could upregulate the expressions of ET-1,eNOS and iNOS mRNA in RMECs and increase the release of ET-1 and NO.This pathway might be one of pathologic mechanisms of retinal injury induced by high intraocular pressure.

Objective To explore the changes of cardiomyocytes apoptosis in the effect of irhesartan and imidapril on regressing cardiac hypertrophy in spontaneous hypertensive rat (SHR), and its mechanism. Methods Thirty 13-week-old SHR were randomly divided into three groups: SHR positive control group, irhesartan treated group(50 mg ?kg-1 ?day -1 ) , imidapril treated group(3 mg ?kg -1 ?day-1 ), and normotensive Wistar-Kyoto rats(WKY) as controls. Blood pressure of rats were monitored periodically. After 15 weeks, all rats were killed and left ventricle weight(LVW) were measured, plasma and myocardium angiotensin Ⅱ concentrations were examined by radioimmunoassay. Then the changes of cardiomyocytes apoptosis using in situ TDT-mediated dUTP nick end labeling(Tunel) were studied. Results Blood pressure, LVW, AngⅡ level of plasma and left ventricle were higher in SHR than those in WKY(P

Purpose To investigate the role of S3I-201 on tubular interstitial lesion in lupus nephritis.Methods MRt/MpJ mice were designated as the control group.MRL/lpr nice were randomly divided into LN group,S3I-201 group and DMSO group.The serum and 24 h-urine were collected to detect the serum creatinine,blood urea nitrogen and urine protein.Immunohistochemistry was used to detect the expression of FN.Western blotting analysis was used to determine the expression of E-cadherin,α-SMA,MCP-1,ICAM1,STAT3 and p-STAT3.Results Compared with the expression level in control group,the protein level of α-SMA,MCP-1,ICAM1 and FN were increased in renal tissue of MRL/lpr mice,while the expression of E-cadherin was markedly decreased.And the STAT3 was activated in renal tissue of MRL/lpr mice.The administration of S3I-201 could inhibite the activation of STAT3 and ameliorate the expression of E-cadherin,α-SMA,MCP-1,ICAM-1 and FN.Conclusion S3I-201 can relieve the tubular interstitial leison,which maybe concerned with the phosphorylation of STAT3.

Objective To investigate the prognostic value of CD 4+CD25+/high CD127 low/-and CD14+HLA-DRlow/-for evaluating the severity of acute pancreatitis .Methods The percentages of CD4+CD25+/highCD127low/-and CD14+HLA-DRlow/-and the CD64 index were measured by flow cytometry in pa-tients with acute pancreatitis ( including 43 cases of mild acute pancreatitis and 24 cases of severe acute pan-creatitis).Moreover, the levels of C-reactive protein (CRP), acute physiology and chronic health evaluationⅡ( APACHEⅡ) score and CT severity index ( CTSI ) were detected for a correlation analysis .Results The percentages of CD4+CD25+/highCD127low/-and CD14+HLA-DRlow/-and the CD64 index in patients with severe and mild acute pancreatitis were significantly higher than those in healthy subjects .Patients with se-vere acute pancreatitis showed higher percentages of CD 14+HLA-DRlow/-than patients with mild acute pan-creatitis.With the disease progression, the CD64 index and the levels of CD4+CD25+/highCD127low/-, CD14+HLA-DRlow/-and CRP were significantly dropped after an initial increase in patients with mild acute pancrea -titis, while these indexes were continuously elevated in patients with severe acute pancreatitis .The percent-age of CD14+HLA-DRlow/-was positively correlated with CD64 index, CRP level, APACHEⅡ score and CTSI.Conclusion CD14+HLA-DRlow/-level was closely related to the severity of acute pancreatitis , which could be used as immune parameter for the estimation of the clinical severity of acute pancreatitis .

Objective To study the correlation between the post-transplant renal allograft function and the variation of serum cystatin C (CyC) concentration in renal allograft recipients. Methods One hundred and ninety-three renal allograft recipients accepted the same combination immunosuppressive regimen of tacrolimus, mycophenolate and prednisone were enrolled into the study. Patient's serum and urine samples were collected on day 5 post-transplant to detect serum cystatin C, serum and urine creatinine (SCr). Correlation analysis was used to analyze correlation between CyC concentration and SCr concentration or the calculated creatinine clearance rate (CkCCr) by using the Cockcroft-Gault equation and urine creatinine clearance rate (CCr). Specificity and sensitivity of using the CyC concentration to evaluate glomerular filtration rate (GFR) were calculated as well.Results The mean concentrations of serum CyC and SCr on day 5 post-transplant were (1.91±1.2)mg/L and (174.0±129.1)μmol/L, respectively. While the CCr and CkCCr were (67.9±27.3)ml/min and (68.1±27.8)ml/min, respectively. Forty-two patients had a CyC concentration below 1.25 mg/L, 102 patients'CyC concentrations were between 1.25 and 2.0 mg/L and 49 patients'CyC concentrations were above 2. 0 mg/L. As for SCr, 62 patients had a concentration below 125 μmol/L, 83 patients'concentrations were between 125 and 200 μmol/L and 48 patients'concentrations were above 200 μmol/L. For CkCCr, there were 52 cases with a concentration above 80 ml/min, 96 cases with a concentration between 80 and 60 ml/min and 45 cases with a concentration below 60 ml/min. Serum CyC concentration had a negative correlation with CkCCr (r=-0. 907, P＜0. 001) and had a significantly positive correlation with SCr concentration (r=0. 886, P＜0. 001). SCr had a significantly negative relationship with CkCCr (r=-0. 889 ,P＜0. 001). Serum CyC had higher correlation with CkCCr than the correlation between SCr and CkCCr. The ROC curves showed that areas under curve of CyC, SCr, CCr and CkCCr were 0. 877, 0. 771, 0. 832 and 0. 909, respectively. Specificity and sensitivity of CyC, SCr,CCr and CkCCr were 69.3%, 96.1%, 77.1%, 71.3%and 91.6%, 52.2%, 67.5%, 84.6%, respectively.Conclusions Serum CyC concentration elevates earlier than SCr concentration when there is slight renal function impairment. Serum CyC concentration might become a more sensitive marker to evaluate the post-transplant renal allograft function in renal transplant recipients.

OBJECTIVETo investigate the feasibility and accuracy of length measurement of in vivo teeth by using cone beam CT (CBCT).

METHODSBefore orthodontic extraction, 109 vital premolars from 40 participants were scanned by using CBCT and reconstructed by using InVivoDental software. Buccal-lingual sectional images along the long axis of teeth were then acquired, and the crown, root, and tooth length were measured separately. After careful extraction and fixation, the corresponding length of the same tooth was measured by using a digital caliper. CBCT measurement accuracy was then verified by using physical measurements as reference.

RESULTSCBCT and the physical method did not obtain significantly different measurements of the root, crown, and tooth length of experimental teeth (P=0.790, P=0.621, P=0.657, respectively), and the measurements were found to be consistent. The 95% limits of agreement of root, crown, and tooth length were -1.10 mm to 1.13 mm, -1.00 mm to 0.96 mm, and -1.00 mm to 1.05 mm, respectively.

CONCLUSIONThe difference between CBCT and the physical method was not significant, and good consistency was shown. CBCT could be applied in noninvasive measurement of in vivo teeth.

Objective To investigate the prognostic value of regulatory B cells (Bregs) in patients with acute pancreatitis .Methods Flow cytometry analysis was performed to detected the percentages of CD19+IL-10+and CD19+CD24hiCD27hi Bregs in peripheral blood samples collected from patients with acute pancreatitis (36 cases with mild acute pancreatitis and 15 cases with severe acute pancreatitis ) as well as the surface costimulatory molecules including CD80 and CD86 on CD19+CD24hiCD27hi Bregs.Their correlations with lymphocytes and C-reactive protein ( CRP) were further analyzed .Results The numbers of lympho-cytes, CD19+lymphocytes, CD19+IL-10+and CD19+CD24hi CD27hi Bregs in peripheral blood samples col-lected from patients with severe and mild acute pancreatitis as well as the mean fluorescence intensities ( MFI) of CD80 and CD86 were significantly lower than those from healthy subjects .Compared with patients with mild acute pancreatitis , the numbers of lymphocytes and CD 19+lymphocytes , the absolute numbers of CD19+IL-10+and CD19+CD24hiCD27hi Bregs as well as the mean fluorescence intensities (MFI) of CD80 and CD86 in patients with severe acute pancreatitis were significantly decreased .The percentages of CD19+IL-10+and CD19+CD24hiCD27hi Bregs in patients with mild acute pancreatitis were significantly increased af-ter an initial drop , but in patients with severe acute pancreatitis those values were continuously decreased along with the disease progression .The percentage of CD19+IL-10+Bregs was positively correlated with the percentage of CD19+CD24hiCD27hi Bregs and the absolute number of CD19+lymphocytes, but was negatively correlated with CRP .Conclusion The abnormal number and function of CD 19+IL-10+and CD19+CD24 hi CD27hi Bregs might be one of the important reasons causing immune dysfunction in patients with acute pan -creatitis.

Objective:To establish an HPLC method to simultaneously determine the content of protocatechualdehyde, rosmarinic acid and salvianolic acid A in Salvia miltiorrhiza Bunge. Methods: The chromatographic column was a Linksil-ODS ( 250 mm × 4. 6 mm,5 μm) column,the mobile phase was 1% acetic acid water solution-acetonitrile (7∶3) with the flow rate of 0. 8 ml·min-1 . The detection wavelength was 280nm , the column temperature was 30℃ and the injection volume was 20 μl. Results: The linear range of protocatechualdehyde, rosmarinicacid and salvianolic acid A was 3-300 μg·ml-1(r>0. 999 0), and the recovery was within the range of 95. 22%-99. 68% with RSD below 2% (n=5). Conclusion:The method is accurate, rapid and reproducible, and can be used to control the quality of Salvia miltiorrhiza Bunge.

Objective To observe the serum levels of calcium, phosphorus, alkaline phosphatase (ALP), parathy-roid hormone (PTH) and 25-hydroxyvitamin D, and the influence of biochemical markers of bone turnover in Graves’dis-ease. Methods Sixty-two patients with Graves’disease were enrolled into the Graves’disease group and 91 healthy indi-viduals as a control group. Electrochemical luminescence was used to evaluate the plasma levels of PTH and 25-hydroxyvita-min D in two groups. The serum levels of calcium, phosphorus and ALP were measured with biochemistry methods in two groups. Results The serum levels of calcium, phosphorus, ALP and 25-hydroxyvitamin D were significantly higher in the Graves’disease group compared with those in control group (P<0.01). The serum levels of calcium, phosphorus, ALP and 25-hydroxyvitamin D were significantly higher in female patients than those of control group, and the level of PTH was lower than that of control group. For male patients, the levels of ALP and 25-hydroxyvitamin D were higher than those of control group, and the level of PTH was lower than that of control group. In Graves’disease group, patients with vitamin D deficien-cy were 17 cases (27.4%), insufficiency 20 cases (32.3%) and sufficiency 25 cases (40.3%), respectively. In control group, there were 54 cases with vitamin D deficiency (59.3%), 31 cases with insufficiency vitamin D (34.1%) and 6 cases with suffi-ciency vitamin D (6.6%), respectively. There was no correlation in plasma levels of PTH, 25-hydroxyvitamin D, serum calci-um and serum phosphorus in Graves’disease group. Conclusion The bone turnover is accelerated in Graves’disease. The increased plasma level of 25-hydroxyvitamin D is related with increased calcium level and decreased PTH level in Graves’ disease. The increased serum phosphorus reduces 1-α-hydroxylase activity. Vitamin D deficiency plays a minor role in bone metabolism of Graves’disease.