Long-term prognosis and quality of life of survivors with hepatitis B virus-related acute-on-chronic liver failure / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the long-term prognosis and health-related quality of life of patients surviving hepatitis B virus-related acute-on-chronic liver failure (HBV-ACLF).</p><p><b>METHODS</b>The clinical data were collected from patients with HBV-ACLF, who were hospitalized in our department between November, 2011 and October, 2016 and survived for more than 90 days. The patients were followed for occurrence of newly diagnosed cirrhosis, decompensation events, hepatocellular carcinoma and death. The quality of life of the patients was evaluated using SF-36 score, and the patients with chronic hepatitis B (CHB) and cirrhosis treated during the same period served as controls.</p><p><b>RESULTS</b>A total of 223 ACLF survivors were included in this study. According to the presence of cirrhosis on admission, the enrolled patients were divided into chronic hepatitis B-related ACLF (CHB-ACLF) group (n=130) and liver cirrhosis ACLF (CIR-ACLF) group (n=93). The 12-, 24- and 50-month survival rates in CHB-ACLF group were 97%, 95.7% and 93.9%, respectively, significantly higher than the rates in CIR-ACLF group (91%, 86% and 74%, respectively; P=0.007). In patients with CHB-ACLF, the 12-, 24- and 36-month progression rates of cirrhosis were 37.9%, 58.4% and 68.7% respectively. Multivariate Cox regression identified the peak value of serum creatinine (HR=1.015, P=0.026) and INR (HR=2.032, P=0.006) within 28 days as independent risk factors and serum sodium at baseline (HR=0.84, P=0.035) as an independent protective factor of occurrence of cirrhosis. The score of mental health on SF-36 in ACLF group was significantly lower than the national norms, and the scores for general health and body pain of ACLF patients were significantly higher than those in patients with CHB or cirrhosis.</p><p><b>CONCLUSION</b>The long-term prognosis of ACLF survivors with and without cirrhosis can be different. Acute attacks are associated with an increased rate of cirrhosis progression in CHB patients who recovered from ACLF, possibly in relation with the severity of extra-hepatic organ injuries. The physical and social functions of long-term survivors of ACLF do not significantly decline, but their psychological status can be affected.</p>

Long-term exposure to PM2.5 from automobile exhaust results in reproductive dysfunction in male rats / 中华男科学杂志

<p><b>OBJECTIVE</b>To explore the effects of long-term exposure to particulate matter 2.5 (PM2.5) from automobile exhaust on the reproductive function of Sprague Dawley (SD) rats.</p><p><b>METHODS</b>Forty-five male SD rats, weighing 80 - 94 g and aged 28 days, were randomly assigned to receive intra-tracheal administration of 0.9% normal saline (control group, n = 15), PM2. 5 at 2 μg per 100 g body weight per day (low-dose PM2.5 group, n = 15), and PM2.5 at 16 μg per 100 g body weight per day (high-dose PM2.5 group, n = 15), qd, for 60 successive days. After the last 24-hour exposure, 10 rats were taken from each group for copulation with normal female ones, while the others were sacrificed, their testes removed for sperm count and deformity, pathological examination, and determination of the Connexin43 expression.</p><p><b>RESULTS</b>The conception rate was significantly decreased in the low- and high-dose PM2.5 groups as compared with that of the control (70% and 50% vs 100%), and so were the sperm count and quality. The rats in the PM2.5-exposed groups showed significantly disordered histological structure of the seminiferous tubules, reduced sperm count in the testicular lumen, some exfoliated secondary spermatocytes, downregulated Connexin43 expression in the testis, and damaged blood-testis barrier.</p><p><b>CONCLUSION</b>Long-term exposure to PM2.5 from automobile exhaust damages the reproductive function of male SD rats.</p>

Construction of a duck hepatitis B virus YMDD mutant and identification of its resistance phenotype / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a lamivudine-resistant plasmid containing 1.2 unit genome of duck hepatitis B virus and identify its replication and drug-resistance in avian LMH hepatica cells.</p><p><b>METHODS</b>The recombinant plasmid PBS-DHBV1.2 was constructed using the 1.2-genome length DHBV DNA sequence from a dimer DHBV genome with pcDNA3.1 as the template. With site-directed mutagenesis, we obtained PBS-DHBV1.2-M512V plasmids with polymerase gene mutation from PBS-DHBV1.2. Two constructed plasmids were transiently transfected into LMH cells using FuGENETM6 transfection reagent and cultured in the medium containing different concentrations of lamivudine. Southern blot hybridization was performed to detect DHBV replication intermediates.</p><p><b>RESULTS</b>PCR amplification, restriction digestion and plasmid sequencing all confirmed successful construction of PBS-DHBV1.2-M512V recombinant plasmid. Southern blot analysis identified the presence of all the expected DHBV replication intermediates in LMH cells. The replication capacity of the mutant plasmid was decreased by 2.7 times compared with that of the wild plasmid. The IC(50) of lamivudine was 37.12∓8.81 ng/ml for the mutant, greater than that of the wild plasmid (10.90∓4.80 ng/ml).</p><p><b>CONCLUSION</b>Compared with the wild plasmid, the mutant plasmid has a lower replication capacity and sensitivity to lamivudine in vitro.</p>

The effect of childbirth on carcinogenesis of DMBA-induced breast cancer in female SD rats / 癌症

Many epidemiologic and clinical studies have indicated that the frequency of breast cancer was lower in parous women than in nulliparous women. Moreover, the incidence of breast cancer has been reported to be lower in women with early childbirth than in women with late childbirth. To verify the effect of childbirth and the age at first childbirth on carcinogenesis and progression of breast cancer, we induced breast cancer by 7,12-dimethylbenanthracene (DMBA) in 120 female Sprague-Dawley (SD) rats, and divided them into control or experimental (DMBA-treated) nulliparous, early childbirth, and late childbirth groups to observe the incidence, latency, and size of breast cancer. Argyrophilic nucleolar organizer regions (AgNOR) count and the expression of C-erbB-2, proliferating cell nuclear antigen (PCNA), Ki-67, and minichromosome maintenance protein 2 (MCM2) in breast cancer tissues were detected by immunohistochemistry. The breast cancer incidences were 95.0%, 16.7%, and 58.8% in the experimental nulliparous, early childbirth, and late childbirth groups, respectively (all P < 0.05). Between any two of these groups, the latency was significantly different, but tumor size was similar. AgNOR count and the expression of C-erbB-2, PCNA, Ki-67, and MCM2 were significantly higher in the experimental nulliparous group than in the experimental early or late childbirth groups (P < 0.05), but no significant differences were observed between the latter two groups. Taken together, the results suggest that childbirth, especially early childbirth, can reduce the incidence and postpone the onset of DMBA-induced breast cancer.

Alteration of thromboxane A2 and prostacyclin levels in rats with obstructive jaundice and the effect of salviae miltiorrhizae on them / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the alteration of plasma levels of thromboxane A2 (TXA2) and prostacyclin (PGI2) as well as changes of microcirculation in renal cortex of obstructive jaundice model rats, and to study the effect of Radix Salviae miltiorrhizae (SM) on them.</p><p><b>METHODS</b>SD rats were randomly divided into three groups: the sham operation group (A), the common bile duct ligation model group (B), and the SM treated model group (C). Their blood plasma TXA2/PGI2 ratio (T/P), blood levels of urea nitrogen (BUN) and creatinine (Cr) were determined respectively in batches (8 rats from each group) on the 3 rd, 7th and 10th day, their capillary caliber (CC) in renal cortex was measured at the same time points using WX-9 type microcirculation microscope.</p><p><b>RESULTS</b>Compared with Group A, T/P was higher and CC was smaller in Group B at all the time points. Levels of BUN and Cr increased on day 7 and day 10 after modeling (P<0.05), and they were increasing markedly along with the elongation of the obstructive time (P<0.05). As compared with Group B, T/P was lower and CC was expanded in Group C, with levels of BUN and Cr lowered on day 10 (P<0.05).</p><p><b>CONCLUSION</b>T/P elevation and renal microcirculation obstacle are the important factors for inducing renal injury in obstructive jaundice, and SM shows a protective effect on kidney against the injury.</p>

Analysis of clinical features and virological characteristics in patients infected with three different HBV subgenotypes / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the clinical characteristics and the pattern of precore and core promoter mutations of hepatitis B virus (HBV) subgenotypes Ba, C1 and C2.</p><p><b>METHODS</b>A cohort of 151 patients with chronic HBV infection in Guangdong province of China was enrolled in this study. HBV subgenotypes were determined by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP). Precore and core promoter mutations were analysed using nucleotide sequencing.</p><p><b>RESULTS</b>Of the 151 patients, 80, 51 and 20 were infected with subgenotypes Ba, C1 and C2 respectively. No significant differences were found in HBeAg positivity and liver functional indexes among these three subgenotypes when age and sex were matched. Virologically, HBV/Ba showed the highest frequency of A1896 mutation but the lowest frequency of T1762/A1764 mutation. HBV/C1 was associated with the highest tendency to develop T1762/A1764 mutation, but the lowest prevalence of A1896 mutation. HBV/C2 was associated with an intermediate tendency to develop A1896 and T1762/A1764 mutations.</p><p><b>CONCLUSION</b>Different mutation patterns in precore and core promoter regions are responsible for HBeAg-negative HBV infections among different subgenotypes.</p>

Changes of serum HBsAg in HBeAg positive chronic hepatitis patients with sustained viral response to long-term lamivudine treatment / 中华肝脏病杂志

<p><b>OBJECTIVE</b>HBsAg loss is rare in chronic hepatitis B patients, even in the patients with long-term nucleos(t)ide analogue therapy; therefore information about serum HBsAg kinetics will be of value in understanding this unusual occurrence.</p><p><b>METHODS</b>Forty-five consecutive patients were studied, which were all HBeAg positive and never had antiviral therapy prior to lamivudine treatment; they then achieved rapid and good viral responses (defined as undetectable HBV DNA [Roche Lightcycler, less than 1000 copies/ml] at treatment week 24 and they remained so until week 156). Abbott Architect HBsAg assay was used to quantify serum HBsAg and HBV genotypes were determined by direct sequencing.</p><p><b>RESULTS</b>Twenty-six (57.8%) patients had HBeAg loss during the observation and one patient had HBsAg loss following his HBeAg seroconversion. Serum HBsAg levels decreased to 39.5% (median) of their baseline values at week 12, but no further significant reductions of serum HBsAg were found afterwards. Changes of serum HBsAg were comparable between patients with or without HBeAg loss. Serum HBsAg levels at their baselines were higher in HBV genotype B (HBV/B, n = 21) patients than in genotype C (HBV/C, n = 24) patients. HBV/B patients achieved many more HBsAg reductions than HBV/C ones (75.5 vs. 26.0%, median, P less than 0.05) in the first 12 treatment weeks, however HBsAg levels at week 156 were comparable between these two subgroups. HBsAg changes mainly showed two distinct patterns: a biphasic pattern (HBsAg levels were less than 60% of baseline ones at week 12 and 24, n = 25) and a maintaining pattern (HBsAg levels were greater than 80% of the baseline ones at week 12 and 24, n = 14). Logistic regression analysis showed that low serum HBsAg at baseline (odds ratio 0.020, 95% confidence interval 0.002-0.743, P less than 0.05) and HBV/C infection (odds ratio 8.206, 95% confidence interval 1.070-62.948, P less than 0.05) were the determinants of the occurrences of the maintaining pattern.</p><p><b>CONCLUSION</b>In patients we examined, their HBsAg changes were mainly presented as either a biphasic pattern or a maintaining pattern, which were associated with HBV genotypes (B/C) but not with HBeAg loss. This might explain that why HBsAg loss is a rare occurrence even with long-term lamivudine therapy.</p>

Kinetics of HBV mutants conferring adefovir resistance (rtn236t) and a method to detect them rapidly / 中华肝脏病杂志

<p><b>OBJECTIVE</b>The aim was to build a PCR-RFLP method for detecting rtN236T mutants and to observe their kinetics in chronic hepatitis B (CHB) patients.</p><p><b>METHODS</b>Seven CHB patients who had suboptimal viral response or viral breakthrough under adefovir mono-therapy were studied. Part of the HBV reverse transcriptional gene from serial sera samples was sequenced with PCR products or cloned HBV DNA; mutations at rt236 were simultaneously analyzed by a PCR-RFLP assay. Genetic diversity of HBV was observed by calculating Hamming distance within domains B, C and D of RT.</p><p><b>RESULTS</b>Three patients had viral breakthrough and one with suboptimal viral response had adefovir-resistance mutants, one had rtA181V mutation and three had rtN236T mutation. A novel PCR-RFLP assay based on restriction enzyme HpaI or DraI for on the detection of rtN236T mutant was established, which detected 10% minor strains with 100% specificity. Mutants (rtA181V or rtN236T) appeared 0-8 months earlier than the viral breakthrough, then afterwards became the dominant ones. In one patient after stopping the adefovir therapy, 3 months later a wild type virus re-took again the mutant one (rtN236T); in one patient who developed a rt236T mutant after 132 weeks of adefovir treatment, a novel mutant (rtN236V) appeared and then became the dominant one while adefovir treatment continued.</p><p><b>CONCLUSIONS</b>A rapid and easy method was established to detect rtN236T mutants. Mutants for adefovir-resistance accumulated rapidly then became dominant, but they could be taken over again by a wild type or novel mutant HBV.</p>

Mutations of HBV polymerase gene sequence in lamivudine-resistant chronic hepatitis B patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the characteristics of mutation in HBV polymerase (P) gene reverse transcriptase region (RT region) in lamivudine-resistant chronic hepatitis B (CHB) patients.</p><p><b>METHODS</b>This study involved 115 CHB patients who developed clinical resistance to lamivudine. Direct sequencing of the PCR products was used to detect lamivudine genotypic resistance.</p><p><b>RESULTS</b>Lamivudine resistant mutation was detected in 103 patients, and the major mutations included rtL180M+rtM204V and rtM204I, accounting for 58.3% and 22.3%, respectively. Other resistant substitutions included rtL80V/I, rtT184S, and rtA200V, and combined mutation of triple resistant substitutions was detected in HBV RT region of 5 patients by direct sequencing.</p><p><b>CONCLUSION</b>For lamivudine-treated patients, combined mutation at the sites other than rtL180 and rtM204 in HBV P gene should also be detected for drug resistance evaluation.</p>

Experimental study on constructing small-caliber artery by tissue engineering approach / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the possibility of constructing small-caliber artery by means of tissue engineering.</p><p><b>METHODS</b>Cell-PGA mixtures were made by separately seeding 1 x 10(7) smooth muscle cells and 5 x 10(6) endothelial cells isolated from neonate umbilicus onto PGA scaffold, the cell-PGA constructs were wrapped around a silicone tube before its implantation subcutaneously to nude mice and the mice were sacrificed in 2 and 6 weeks. The tissue engineered artery (TEA) were examined both grossly and immunohistochemically.</p><p><b>RESULTS</b>The gross appearance of TEA was similar to that of the natural counterparts; histologic and immunohistochemical analyses of the neoformed tissues revealed a typical artery structure, including the presence of EC at the luminal surface and the presence of SMC and collagen in the wall.</p><p><b>CONCLUSION</b>TEA with histology similar to natural vessel can be constructed by tissue engineering.</p>