AIM:To investigate the expression of TLR2 and TLR4 in hepatocellular carcinoma(HCC),and to analyze their correlations to clinicopathologic features of HCC.METHODS:The protein and mRNA levels of TLR2 and TLR4 in HCC and para-tumor tissue were determined by immunohistochemistry and real-time fluorescence quantitative PCR(RFQ-PCR).RESULTS:The protein and mRNA levels of TLR2 and TLR4 in HCC were lower than those in para-tumor tissue(P

Objective To study and analyze the cause, prevention and treatment for complications in patients with serious hypospadias repaired by one-stage urethroplasty. Methods From 1987 to 2002,275 patients with serious hypospadias were repaired by one-stage urethroplasty, there were 35 cases had complications. The classifications were penoscrotal 148 cases, scrotal 95 cases and perineal 32 cases. The lengths of new urethras were from 3.0 to 8.4 cm, the mean was (4.1?0.7) cm. Thirty-two cases were received endocrinotherapy before urethroplasty. Results The rate of complications was 12.7% for 1～3 years following survey. There were urethral fistulas 24 cases (8.7%),urethral strictures 6 (2.2%), diverticulums 3 (1.1%), chordees 2 (0.7%).The rate of urethral fistulas was the first and urethral stricture was the second, they were higher than those of other complications (P

【Objective】 To study the relationship of pathologic changes and serologic markers of fibrosis in patients with viral hepatitis. 【Me thods】 Liver s pecimens were obtained by percutaneous needle biopsy under color Doppler ultras ound guidance in 299 patients with viral hepatitis. The specimens were stained b y hematoxylin and eosin (HE), Gordon and Sweet's reticulum methods (RT), in orde r to determine the degree and the stage of pathologic changes with microscopy. H yaluronic acid(HA), collagen type Ⅳ(Ⅳ-C) and human precollagen type Ⅲ（HPCⅢ ）as serum fibrous markers were detected by radioimmunoassay. 【Results】 The se rum levels serologic markers were slightly increased in 97 patients with mild ch ronic hepatitis, moderately increased in 126 patients with moderate chronic hepa titis, and significantly increased in 29 severe cases and 47 subjects with cirrh osis. Both the grade of inflammatory activity and the stage of fibrosis were clo sely related to the levels of serum fibrous markers. 【Conclusion】 Chronic vira l hepatitis pathologic feature and levels of serum markers of fibrosis change al ong with clinical process of patients. The combination of liver biopsy and detec tion of serum markers of fibrosis might be highly valuable for the diagnosis.

Objective To observe the expression of TLR4 in hepatocellular carcinoma cell line HepG2 after transient and stable HBV genome transfection. Methods Immunofluorescence flow cytometry was used to detect mean fluorescence intensity (MFI) of TLR4 and TLR4 positive cell percentage in hepetocellular carcinoma cell lines HepG2 and HepG2. 2.15. Various doses of HBV DNA plasmid were transfected into HepG2 cells with lipefectamine 2000. Immunofluoroscenee flow cytometry was used to detect MFI of TLR4 and TLR4 positive ceil rate of infected HepG2 ceils. Trypan blue staining was used to examine the sum of living cells. Results MFI of TLR4 and TLR4 positive cell rate of HepG2.2.15 cells were significantly higher than those in HepG2 cells (both P' <0. Ol). MFI of TLR4 and TLR4 positive cell rate of HepG2 cells transfected by various doses of HBV DNA were significantly higher than those in control group (all P' < 0. 01). MFI of TLR4 and TLR4 positive cell rate of infected HepG2 cells were positively correlated with the doses of HBV DNA (both P' <0. 01) and negatively correlated with the sum of living cells (both P' <0. 01). Conclusions Enhanced expression of TLR4 appeared in HepG2 cells with both transient and stable HBV infection, along with reduction of living cells.

Objective To investigate HBV genotypes and gene mutations in chrenic hepatitis B (CHB) patients with liver failure after lamivudine withdrawal. Methods Twenty four patients with relapsing CHB after lamivudine withdrawal were divided into liver failure group ( n = 12 ) and chronic hepatitis group ( n = 12 ). HBV DNA from these patients was amplified by PCR. The PCR products were cloned into PGEM-T vector and HBV DNA sequences were analyzed. Results In liver failure group, there were 6 sequences detected, in which 3 were of genotype B and 3 were of genotype C. In chronic hepatitis group, there were 9 sequences detected, in which 2 were of genotype B and 7 were of genotype C. Compared with the wild type HBV sequences, there were multiples mutations in S, P, C, X regions. Gene mutations in high conservative sequences of BCP and P regions were detected in liver failure patients after lamivudine withdrawal. Conclusions In HBV patients with liver failure after lamivudine withdrawal, half of them were of genotype B and the others were of genotype C. Some mutations in high conservative sequences of BCP and P regions may be related to the liver failure in these patients.

98%. In the pDOR-myb infected HSCs, c-myb expression levels, the cell proliferation, and ? 1-Ⅰ collagen mRNA expression were repressed significantly. CONCLUSIONS: c-myb plays a key role in the activation and proliferation of HSC. c-myb antisense RNA can inhibit cell proliferation and ? 1-Ⅰ collagen mRNA expression in the infected HSC. These data suggest that inhibition of c-myb gene expression would be a potential way for the treatment of liver fibrosis.

ObjectiveTo investigate the characteristic parameters and variation of acoustic startle reflex (ASR) and conditioning response (CR)during the acquisition of delay-trace eyeblink conditioning.Methods15healthy Kunming mice were randomly divided into 3 groups:single-task DEBC group ( n =5 ) ; single-task TEBC group ( n =5) ; dual delay/trace group ( n =5).Three groups received paired training of tone conditioned stimulus(CS)in different audio frequency (TEBC groups:2KHz; DEBC group:1KHz) and a 100 ms comeal oxygenpuff unconditioned stimulus(US).Then observed the characteristic parameters of ASR and CR.ResultsAfter pairing training for 10 days,ASR rate( ASR％ ) of single-task groups ( 10th day (38.00 ± 8.64) ％ and (38.55 ±12.41 ) ％ respectively,time effect:F =4.574,P =0.008 ; group effect:F =2.021,P =0.193 ) and dual delaytrace group ( 10th day (47.95 ± 14.23 ) ％ and (62.01 ± 9.03 ) ％ respectively,time effect:F =5,547 P =0.013 ;group effect:F =0.738,P =0.415) changed significantly with the following training but between single-task groups or dual delay-trace group.CR rate ( CR％ ) of single-task groups ( 10th day were (45.4 ± 5.39) ％ and (65.6 ± 6.77) ％ respectively,time effect:F =9.558,P =0.000 ; group effect:F =5.117,P =0.054 ) and dual delay/trace group ( 10th day (57.66 ± 4.34) ％ and (77.35 ± 7.36) ％ respectively,time effect:F =7.750,P =0.002 ;group effect:F=1.449,P=0.263 ) showed obvious change with the following training but between singletask groups or dual delay-trace group.Peak amplitude of single-task DEBC group ( time effect F =2.679,P =0.017 ) and dual delay-trace group ( F=3.452,P=0.034 ) had increased significantly with the following training.Peak latency showed no obvious change.ConclusionDual delay-trace conditioning procedure of classical eyeblink conditioning and single-task can be successfully established in Kunming mice.Although there are some variations,those can be controled and be weaken effect of them by taking targeted measures.

【Objective】To construct the c-myb antisense RNA recombinant retroviral vector and its packaging cell line.【Methods】The segment of c-myb gene was cloned into pUC19 with TA cloning method after amplication by RT-PCR,and then was subcloned into retroviral vector pDOR.The recombinant retroviral vector named pDOR-myb was transfected into retroviral package cell line PA317 after selection with G418.【Results】Sequencing data indicated that the c-myb gene was exactly identical to the sequence in the GenBank.The segment of c-myb gene was inserted directionally into pDOR.Resistant colonies were obtained and the titers of pDOR-myb were 5.2×104～9.5×104 CFU/mL.【Conclusion】The recombinant retroviral vector containing c-myb gene is successfully constructed and its packaging cell line PA317/pDOR-myb was established.

【Objective】 To investigate the penile appearance, sexual function, psychological status and related influencing factors of adult patients who underwent hypospadias repair surgery in their minors, so as to provide reference for the diagnosis, treatment and prognosis of hypospadias. 【Methods】 This study included 50 adult hypospadias patients who underwent urethroplasty in our hospital during May 2005 and Aug. 2018. The present appearance, sexual function and psychological status were evaluated. The correlation and consistency between hypospadias objective scoring evaluation (HOSE) and pediatric penile perception score (PPPS) were analyzed. Factors affecting the results were determined with univariate and multivariate regression analysis. 【Results】 The satisfaction rate of HOSE was significantly correlated with the urethral length and complications (P=0.024, P=0.033). The satisfaction rate of PPPS was significantly correlated with the number of urethral operations and postoperative complications (P=0.041, P=0.023). There was a weak correlation between HOSE and PPPS (r=0.291, P=0.040), but almost no consistency (Kappa=0.2, P=0.107). Sixty percent of the patients paid attention to the ventral appearance of penis, whose dissatisfaction rate of PPPS was higher than those who did not pay attention to the ventral appearance of the penis (19/30 vs. 6/20, P=0.021). Patients with multiple operations (>1), postoperative complications or unsatisfactory penile appearance were more likely to have inferiority complex (52.6% vs. 22.6%, P=0.029; 59.1% vs.14.3%, P=0.001;61.5% vs. 24.3%, P=0.015). Multivariate regression analysis showed that dissatisfaction with the appearance of the penis was an independent risk factor for patients’ perception of their own physical defects. Among the patients who believed that they had physical defects, the percentage of patients with grade IV penile erectile hardness was significantly lower than that of those who denied they had physical defects (9/17 vs. 27/33, P=0.047). 【Conclusion】 The undesirable postoperative penis appearance is likely to have a negative impact on patients’ long-term psychological state, which might further damage the sexual function in adulthood. Surgeons should pay attention to the penile appearance during the conduction of hypospadias repair.