Objective To explore the influence of XTP4 on genomic expression profile of hepatocyte through screening and cloning of genes trans-regulated by XTP4-expressing plasmid. Methods The expressive vector pcDNA3.1(-)-XTP4 was constructed by routine molecular biological methods, and suppression subtractive hybridization (SSH) method was employed to detect the mRNA differentially expressed by the HepG2 cells transfected with pcDNA3.1(-)-XTP4 and pcDNA3.1(-), respectively, using lipofectamine. The twice enriched PCR products were subcloned into T/A vectors to set up the subtractive library. Amplification of the library was carried out in E. coli strain JM109. The screened cDNA were sequenced and analyzed in GenBank with Blast search after PCR. Results The amplified subtractive library containd 21 positive clones. Colony PCR analysis showed that there were 16 clones containing 200-1000bp inserts. Sequence analysis was performed and 9 kinds of encoding sequences were achieved. These genes trans-regulated by XTP4 protein involved in hepatic fibrogenesis, tumorgenesis, mitochondrial function, and cell growth regulation. Conclusions The findings obtained by SSH provide significant data for a preliminary understanding of the biological function of a new identified gene-XTP4. These results will pave the way for the study of the molecular mechanism of the transactivating effects of HBxAg and the development of new therapy for chronic hepatitis B.

In order to observe the feature of age-related marrow conversion and maturation of epiphyseal cartilage and analyze the distribution of red and yellow marrow in the proximal femur at STIR MR imaging, STIR and T(1) weighted MR imaging of the proximal femur in 52 subjects, aged 4 months to 25 years old, were retrospectively analyzed for the distribution and appearance of red and yellow marrow. The subjects with no known bone marrow abnormalities were divided into 6 age groups. The signal intensity of the marrow in the proximal epiphysis, proximal metaphysis, proximal diaphysis, distal diaphysis and greater trochanter was compared with the signal intensity and homogeneity of surrounding muscle and fat and graded by two observers. The results showed that the conversion of hematopoietic marrow in the proximal femur followed a well-defined sequence, occurring first in the proximal epiphysis, followed by the distal diaphysis, and then greater trochanter and metaphysis. STIR in combination with T(1)-weighted imaging could display clearly the origin of ossification center and the course of conversion from red to yellow marrow in proximal epiphysis and greater trochanter. STIR imaging showed that the marrow conversion in proximal metaphysic began below epiphyseal plate and intertrochanter. The site of red yellow was distributed in weight-bearing axis by 20 years of age. The marrow conversion of diaphysis was from distal end to proximal end, and the consequence of conversion was that distal diaphysis contained yellow marrow but proximal diaphysis partly red marrow connected with the red marrow of metaphysic. The epiphyseal cartilage had different characters of signal-intensity with age in STIR sequence. The distribution of red marrow in STIR imaging was more close to that of anatomy than T(1)-weighted imaging. It was concluded that STIR could dynamically display the feature of marrow conversion and the development of epiphyseal cartilage and accurately reveal the age-related distribution of red and yellow marrow on STIR imaging in the proximal femur.
Age Factors ; Bone Marrow/*anatomy & histology ; Bone Marrow/physiology ; Epiphyses/anatomy & histology ; Femur/*anatomy & histology ; Image Enhancement/*methods ; Magnetic Resonance Imaging/*methods ; Retrospective Studies ; Young Adult

Objective To evaluate the value of reconstruction of three-dimentional CT in development dislocation of hip(DDH)in children.Methods Twelve cases of DDH concluded 4 bilateral and 8 unilateral cases.To sum up,16 sick hips were operated and 8 normal hips were also obtained by three-dimensional CT(Hip speed Fi/x,GE Co).Results 3D reconstruction were used to show femoral head,(acetabulum) and relationship of acetabulum and femoral head respectively.The difference between FNA measurement of sick hips and those of normal hips were significant(P

Objective To investigate the clinical epidemiologic features of transient synovitis(TS) of hip in children occurred in Shen-zhen district.Methods The medical files were reviewed and a standard questionnaire was filled according to the conditions of 705 cases such as pathogeny,clinical manifestation,therapy and prognosis.Results Transient synovitis occurred in a sporadic form all the year round.The peak age of patients with TS was 3-7 years old.The ratio of boys to girls was 2.9:1.About 19.3% patients were attacked an upper respiratory tract infection and 11.9% patients attributed the symptoms to trauma or severe activities before 1 week.A varying degree of painful limp and restriction of movement at the hip were found clinically.All of cases were cured by skin traction.The incidence of recurrence was 6.95%.Conclusions Male predominance is found in TS.It is characteristic of sporadic form in the 4 seasons and intently relation to an upper respiratory tract infection and trauma or severe activities.TS is recurrent and the prognosis is good by skin traction.

Objective:To investigate the purification of antibacterial effective fraction of Syringae folium by macroporous resins. Methods:Static adsorption and desorption tests were carried out to screen the macroporous resins. The desorption experiment was per-formed on the selected D101 resin to optimize the separation process. The effects of resin amount, diameter length ratio, elution flow rate, elution solution concentration and volume were studied. Results:The optimal conditions were as follows:the elution solution was 55% ethanol, the adsorption flow rate was 1 BV·h-1 , the elution flow rate was 5 BV·h-1 , 6 BV 25% ethanol was used to eliminate impurity and 8 BV 55% ethanol was used to elute to obtain the effective fraction. Conclusion: The content of antibacterial effective component is above 65% after purified by D101 resin, indicating that the present method is suitable for large-scale preparation of anti-bacterial effective fraction of Syringae folium.

Objective To investigate the clinical characteristics and risk factors of bacterial liver abscess (BLA) complicated with septicemia.Methods Fifty two BLA patients complicated with septicemia admitted in our hospital from January 2011 to December 2015 were retrospectively reviewed;and 52 cases of BLA without septicemia admitted at the same period were randomly selected as control group.The clinical manifestations, laboratory and radiographic findings, clinical outcome of these patients were analyzed.Logistic regression analysis was used to study the clinical features and risk factors of BLA complicated with septicemia.Results Compared to the control group, the BLA with septicemia group had higher prevalence rates in diabetes mellitus, malignant tumors, jaundice, albumin <35 g/L, BUN≥8.2 mmol/L, hyperglycemia, multiple abscesses and abscesses size ≥10 cm(P<0.05 or <0.01).The blood culture showed that K.pneumoniae(63.3%) was the most commonly isolated pathogen, followed by E.coli(16.7%).Univariate analysis revealed that diabetes mellitus(OR=2.200,95%CI 1.042-4.646), malignant tumors (OR=3.667,95%CI 1.023-13.143), albumin <35 g/ L(OR=2.800,95%CI 1.009-7.774), BUN≥8.2 mmol/L(OR=3.167,95%CI 1.265-7.929), hyperglycemia(OR=3.400,95%CI 1.254-9.216), multiple abscesses(OR=2.667,95%CI 1.043-6.815), abscesses size≥10 cm (OR=5.000,95%CI 1.096-22.820) were positively associated with bacterial liver abscess complicated with septicemia.Multivariate Logistic regression showed that abscesses size≥10 cm (OR=14.016,95%CI 1.354-145.070) was an independent risk factor for complication of with septicemia.Conclusion septicemia is a common complication for bacterial liver abscess, clinically effective measures shauld be taken to prevent and control risk factors associated with septicemia.

Objective To establish a method for detecting HBV cccDNA in hepatocytes of chronic hepatitis B patients.Method 21 liver biopsies from the hepatic operation patients in the hospital of jiangsu province,concluding 19 HBV chronic infected patients (10 HBeAg positive patients and 9 HBeAg negative patients) and 4 uninfected patients,HBV DNA(+) serum of hepatitis B patients was thought as rcDNA.To use proteinase K to release HBV cccDNA and genomic DNA,then divide the cell lysis solution into two parts,one for detecting HBV cccDNA,the other for detecting the number of ?-Globin as internal control. Nucleic acid for detecting HBV cccDNA extracted by phenol-chloroform was digested by plasmid-safe ATP dependent DNase which was applied to digest the single strand DNA in rcDNA and ssDNA,then was quantitated by the primers spanning across the nick and SYBR Green Ⅰ dye.The specifity of PCR production was confirmed by the sequence analysis and rcDNA comparison.The significance of the difference of HBV cccDNA level between HBeAg(+) and HBeAg(-) group was analyzed by two group t test.Results The agarose gelelectrophoresis showed the molecular weight of the PCR production was about 350bp.The coincidence rate of PCR production and goal fragement was nearly 99% by sequence analysis.The result of PCR detection of rcDNA group was negative.The positive rate of HBV cccDNA of liver biopsies of HBeAg (+) patients detected by this method was 100%,the level of HBV cccDNA in the liver biopsies of HBeAg (+) patients was higher than HBeAb(+) patients.Conclusions The specificity of the method is proved by agarose electrophoresis,gene sequencing of the PCR product and rcDNA comparison.The quantitative method that use SYBR Green Ⅰ dye and ?-Globin as internal control is more specific,sensitive and economical,and more suitable for clinical purpose.

Objective To evaluate the expression of Toll-like receptor(TLR)on the monocyte- derived dendritic cells(DC)from chronic hepatitis B(CHB)patients and to analyze the expression pro- file and significance of the TLR such as TLR3,TLR4,TLR?,TLR8 and TLRg,which are associat- ed with immune response to viral infection.Methods Peripheral blood mononuclear cell(PBMC) centrifugated by the hydroxyethyl starch(HES)centrifugation were cultured and induced into DC by granulocyte-maerophage colony stimulating factor(GM-CSF)and interleukin-4(IL-4),and their mor- phology and phenotype were detected by the inverted microscope and flow cytometry respectively. Monocyte-derived DC were obtained from 10 chronically hepatitis B virus(HBV)-infected patients and 15 healthy volunteers.TLR3,TLR4,TLR7,TLRS,TLR9 expression on immature and mature DC were analyzed by FACS Calibur.DC was pulsed with HBcAg on day 3 and 5,then DC maturation and ability to process HBcAg and to stimulate autogeneic T cells were evaluated.Results Monocyte- derived DC developed different TLR expression patterns as they went through different maturation stages.TLR7,TLR8 expressions on immature DC and TLR3,TLR7 expressions on mature DC were lower in CHB than in control(for TLR7,TLR8 expression on immature DC:75.9%,1.0%vs 98.4%,15.4%,P

Objective:To investigate the therapeutic efficacy of double-J catheter in treatment of renal tuberculosis(TB)and in rescuing the structure and function of the kidney.Methods:Thirty-four patients with renal TB(22 combined with single side hydronephrosis)were divided into 2 groups randomly.Group A were treated with antituberculous therapy and group B with antituberculous therapy combined with pre-treatment with double-J catheter.All 34 patients were followed up for 3 months and were re-examined.Results:The results of B ultrasound,intravenous urogram(IVU),CT and isotope nephrogram were comparable between the 2 groups before treatment,and the results were significantly different between the two groups after 3 months'drug treatment(P

Polymerase chain reaction was employed to amplify the whole HBV CP region from the serum of patients with chronic hepatitis B virus(HBV) infection, and then the PCR products were subcloned into pGEM Teasy vectors. Clones were randomly selected to be sequenced and the selected clones were compared to look for the difference.The sequencing results suggested that each sequence of selected clones was different and there were HBV quasispecies groups in patients. There were hot deletion region and point mutation near the TATA like box of CP gene. To address whether the mutations were responsible for the transcriptional activity, the wild type(wt) and the mutants of HBV CP genes were subcloned into pcDNA3 1( ) vectors, respectively. The reverse oriented clones were digested with KpnI and XhoI, and cloned into the KpnI and XhoI sites of the chloramphenicol acetyltransferase (CAT) expressing vector (pCAT3 basic).The recombinant CAT plasmids were transfected into HepG2 cells using lipofectamine PLUS reagent, and the CAT expression which indirectly represented the transcriptional activity of HBV CP lying upstream of CAT gene was detected with a CAT ELISA kit. The restriction enzyme digesting results indicated that the recombinant CAT plasmids were successfully constructed, and the transfection tests indicated that the transcriptional activity of the mutants with deletion or substitute point mutation of TATA like box were reduced in comparison with that of CPwt. The HBV CP gene heterogeneity downregulated the transcriptional activity to some extent.