OBJECTIVE: To evaluate the efficacy of combination therapy of lamivudine (LAM) and adefovir dipivoxyl (ADV) for patients with hepatitis B-induced decompensated cirrhosis. METHODS: A total of 81 patients were randomly divided into a combination group and an ADV group over 48 week treatment course. The combination group were treated with LAM (100 mg/d) plus ADV (10 mg/d), and the ADV group with ADV (10 mg/d ) for 48 weeks. All patients received hepatic function support and symptomatic treatment. The levels of HBV DNA, liver function, Child-Pugh scores and HBV DNA indicators were observed before and after the treatment. RESULTS: At week 4, the mean reduction of HBV DNA was 1.83 lgIU/mL, 17.9% of the patients achieved undetectable HBV DNA and 28.2% showed normal ALT in the combination group. The counterpart in the ADV group was 0.96 lgIU/mL, 5.3% and 10.5%. At week 4, 12, 24 and 48, the differences in the mean reduction of HBV DNA, undetectable HBV DNA and ALT normalization were statistically significant between the 2 groups. The difference in HBeAg negative conversion rates and HBeAg seroconversion at week 24 and 48 was not significant. CONCLUSION The combination therapy results in HBV suppression and improved liver function and Child-Pugh score. The combination treatment has an advantage over ADV due to low drug resistance rate and good tolerance.
Adenine ; administration & dosage ; analogs & derivatives ; Adult ; Antiviral Agents ; therapeutic use ; Drug Therapy, Combination ; End Stage Liver Disease ; drug therapy ; etiology ; Female ; Hepatitis B, Chronic ; complications ; Humans ; Lamivudine ; administration & dosage ; Liver Cirrhosis ; drug therapy ; etiology ; physiopathology ; Male ; Middle Aged ; Organophosphonates ; administration & dosage

OBJECTIVE: To investigate the inhibitive effect of E1A gene carried by PEI-Fe(3)O(4) nanometer particle (NP) on the cell growth of human cervical carcinoma cell in vitro and its mechanism, and to provide the experimental evidence for the feasibility of gene therapy for human cervical carcinoma. METHODS: E1A gene conjugated to PEI-Fe3O4 NP was transfected into human cervical carcinoma cell line Hela. The cell growth curve of Hela was drawn, the doubling time and the number of colony formations on the soft agar were calculated based on the cell count. RT-PCR and Western blot were used to detect the expression of the E1A and HER-2/neu in Hela cells. RESULTS: The cell doubling time of Hela cells transfected with E1A gene (Hela-E1A) was 1.53 times and 1.58 times longer than that of the Hela transfected with blank vector (Hela-vector) and blank Hela control (Hela), respectively. The E1A transfected Hela cells grew slower than those of the control group. The cell colony formation efficiency in the Hela-E1A (6.62%) group was significantly lower than that of Hela (30.48%) and Hela-vector (28.3%) groups (P<0.05). As compared to Hela and Hela-vector, the inhibition rate of Hela-E1A was 78.28% and 76.62% respectively. RT-PCR and Western blot demonstrated that the overexpression of E1A through gene transfection significantly inhibited mRNA and protein expression of HER-2/neu in Hela cells. CONCLUSION E1A gene can suppress the cell growth of human cervical carcinoma cell Hela in vitro. Down-regulated expression of HER-2/neu gene by E1A overexpression in Hela might contribute to the Hela growth inhibitive effect of E1A.
Adenovirus E1A Proteins ; genetics ; Cell Proliferation ; Female ; Gene Expression Regulation, Neoplastic ; Genetic Therapy ; methods ; HeLa Cells ; Humans ; Transfection ; Uterine Cervical Neoplasms ; genetics ; therapy

[ ABSTRACT] AIM:To investigate the changes of migration of lung adenocarcinoma cells promoted by IL-8 and the inner and outer mitochondrial membrane dynamic changes during this process.METHODS:Human lung adenocarci-noma cell line A549 was divided into control group and IL-8 group.Cell migration was analyzed by scratch detection and Transwell assay.The secretion of endogenous IL-8 was detected by ELISA.The protein levels of mitochondrial cytochrome C ( Cyt C) and mitochondrial outer membrane protein Tom20 was detected by Western blot.The mRNA expression of mito-chondrial fusion genes Mfn1, Mfn2 and OPA1 and fission genes Fis1, Drp1 and MTP18 was detected by RT-PCR.The morphological changes of mitochondria were observed by MitoTracker Red CMXRos dye staining and confocal microscopy. RESULTS:The migratory rate of A549 cells and endogenous secretion of IL-8 in A549 cells were higher than those in SPC-A-1 cells.The migratory rate of A549 cells was improved by IL-8 in a time-dependent manner.Compared with control group, the Tom20 protein expression was increased ( P<0.05 ) , and the Cyt C protein expression was decreased ( P<0.05).The expression of mitochondrial outer membrane fusion genes Mfn1 and Mfn2 was increased (P<0.05), and the expression of mitochondrial inner membrane fusion gene OPA1 was decreased (P<0.05).The expression of fission genes Drp1 and MTP18 were decreased (P<0.05), while the expression of Fis1 was no change (P>0.05).Under confocal microscope, the punctate aggregates in the mitochondria of the A549 cells treated with IL-8 were observed.CONCLU-SION:The migratory rate of A549 cells is increased by IL-8, which is related to the changes of mitochondrial fusion genes and the fission genes.

Background:The prevalence rate of diabetes mellitus ( DM ) associated with gastroesophageal reflux disease ( GERD)is increasing recently,and the relationship between DM and GERD has become a research hotspot. Aims:To study clinical features and influential factors of therapeutic efficacy of reflux esophagitis( RE)in elderly patients with type 2 DM(T2DM). Methods:Two hundred elderly patients with RE diagnosed by gastroscopy from March 2011 to October 2013 were enrolled,and divided into T2DM associated with RE( T2DM)group and RE group. The clinical features, endoscopic findings and therapeutic efficacy of the two groups were analyzed,and the influences of DM course and control of blood glucose on efficacy of RE were also analyzed. Results:The main manifestations of RE were extraesophageal symptoms,typical esophageal symptoms were less common. The overall efficacy rate decreased with the increase of endoscopic grade of RE,however,extraesophageal symptoms improved significantly. With the prolonging of DM course,the endoscopic grade of RE was increased,and the control of blood glucose was worse. The overall therapeutic efficacy was significantly reduced with the prolonging of DM course in patients with well controlled blood glucose. Conclusions:Extraesophageal symptoms are common,endoscopic grade of RE is worse and blood glucose is difficult to control in elderly T2DM patients associated with RE. DM course and status of blood glucose control have influences on the therapeutic efficacy of RE.

AIM: To detect the expression level of telomere regulating factor tankyrase and its clinical significance in the origin and development of gastric adenocarcinoma. METHODS: Quantitative Real-time PCR was used to measure the expression of tankyrase mRNA in cancerous and normal adjacent tissues from 16 patients with gastric adenocarcinoma. Immunohistochemistry was used to detect tankyrase protein expressions in 37 gastric adenocarcinoma samples and 13 normal controls. RESULTS: The expression of tankyrase mRNA in gastric adenocarcinoma [median 1.44?10~ -2, range (3.88?10~ -5)-0.4847] was significantly higher than that in normal adjacent mucosa [median 1.0134?10~ -2, range 0-(4.933?10~ -2)] (P

OBJECTIVETo obtain the cDNA sequence encoding fibrinolytic enzyme from Eupolyphaga sinensis and express it in prokaryotic and eukaryotic expression system.

METHODThe primers were designed according to the cDNA of other animals'fibrinolytic enzyme. The cDNA sequence was cloned by RT-PCR and 3 RACE.

RESULTSequence analysis revealed that the length of the cDNA fragment was 672 bp and encoded a protein of 224 amino acid residues, the N end amino acid sequence residues was IVGG in accordance with other fibrinolytic enzyme. The cDNA sequence was expressed in E. coli, inactive protein was obtained. While expressed in Pichia pastoris, recombinant protein had fibrinolytic activity.

CONCLUSIONThe cDNA sequence of fibrinolytic enzyme from E. sinensis Walker was cloned and expressed for the first time and it proved a good basis for further functional study of the enzyme.

Amino Acid Sequence ; Animals ; Base Sequence ; Cloning, Molecular ; Cockroaches ; chemistry ; enzymology ; genetics ; DNA, Complementary ; chemistry ; genetics ; Fibrinolysin ; chemistry ; genetics ; metabolism ; Gene Expression ; Insect Proteins ; chemistry ; genetics ; metabolism ; Molecular Sequence Data ; Sequence Alignment

OBJECTIVETo establish an accurate, fast and simple screening method for AZF microdeletions using capillary technology and use it for clinical testing.

METHODSFor each pair of primers, the 5' end of either forward or reverse primer was labeled with a FAM, JOE or TAMRA fluorescence dyes to establish multiplex quantitative fluorescence PCR systems for the establishment of a screening method of Y chromosome AZF microdeletions by capillary technology. The detection of Y chromosome AZF microdeletion was carried out on 725 cases of non-obstructive azoospermia, oligospermia or asthenospermia.

RESULTSA screening method for Y chromosome AZF microdeletions using capillary technology was established. Thirty eight cases of AZF microdeletions were found among 725 cases of non-obstructive azoospermia, oligospermia or asthenospermia, which gave a deletion rate of 5.24%. Y chromosomal microdeletions were found in 8.62% of the azoospermia group, 6.75% of the oligozoospermic group, and 2.23% of the asthenospermia group.

CONCLUSIONAn accurate, fast and simple screening method of Y chromosome AZF microdeletions by capillary technology has been established, which may have an important clinical value.

Adult ; Azoospermia ; genetics ; Capillary Action ; Chromosome Deletion ; Chromosomes, Human, Y ; Humans ; Infertility, Male ; Male ; Multiplex Polymerase Chain Reaction ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; diagnosis

Objective To investigate the clinical characteristics and risk factors of secondary infection in patients with anti-neutrophil cytoplasmic antibody-associated vasculitis (AAV).Methods One hundred and eighteen patients newly diagnosed with AAV at the institute of nephrology,Tongji hospital affiliated to Huazhong university of science and technology,from 2012 to 2017,were analyzed retrospectively.Induction therapy included single corticosteroids,combination of corticosteroids with cyclophosphamide and combination of corticosteroids with other immunosuppressive agents.End point was defined as moderate to severe infection which was diagnosed by the clinical and radiological manifestation as well as microbiological evidences.The infection-related survival curve was drawn to reflect the time when the infection occurred.The clinical baseline variables in patients with and without infection were compared.Multivariate Logistic regression model was used to determine the independent predictors of infection.Receiver-operating characteristic curve (ROC) was plotted for evaluating the predictive value of lymphocyte on moderate to severe infection.Results During followup of median 3 months (1-30 months),88 infection episodes were found in 63 (53.4％) patients,of which 54 times (61.4％) occurred within 6 months after treatment,46 times (52.3％) happened within 3 months after treatment.The most common organ of infection was lung (62.5％),and the most common pathogen was bacteria (51.1％).Multivariate Logistic regression model showed that lung involvement (OR=4.44,95％ CI 1.59-12.41),moderate reduction of lymphocyte in follow-up (OR=5.69,95％ CI 2.05-15.85) and severe lymphocyte reduction (OR=36.28,95％CI 3.45-381.17) were independent risk factors of secondary infection in AAV patients (all P ＜ 0.05).ROC curve showed that the area under the curve of lymphocyte as a predictor of severe infection was 0.767 (95％ CI 0.64-0.89,P ＜ 0.05).Based on lymphocyte less than 0.49× 109/L which was the cut-off value for predicting severe infection,the sensitivity and the specificity were 83.9％ and 71.9％,respectively.Conclusions Lung involvement and moderate-severe lymphopenia during follow-up are independent risk factors of secondary infection in AAV patients.Hence,physician should pay more attention to those patients,and adjust treatment in time to avoid the occurrence of infection.

Objective:To observe the natural course of patients with simple traumatic intraparenchymal contusions and hematomas (TIPHs), and analyze the risk factors for TIPHs progression.Methods:Using a prospective observational study, 69 patients with TIPHs in Affiliated Xiaolan Hospital, Southern Medical University from July 2018 to January 2020 were selected. The gender, age, cause of injury, compound injury status, combined injury and Glasgow coma score (GCS) were recorded. The plasma levels of prothrombin time (PT), activated partial thromboplastin time (APTT), thrombin time (TT), fibrinogen (Fbg), international normalized ratio (INR), D-dimer, platelet, hemoglobin at admission were detected. The occurrence of hypoxemia and high intracranial pressure were observed. The time of the first CT examination, volume of the first CT hematoma, time of the control CT examination, volume of the control CT hematoma, and lesion morphology, multifocality, subarachnoid hemorrhage (SAH), edema zone and cortical distance showed in the first CT examination were recorded. The risk factors of progression in patients with TIPHs were analyzed.Results:Among 69 patients with TIPHs, TIPHs progression was in 28 cases (progression group), and the progression rate was 40.58%; TIPHs progression was not in 41 cases (non-progression group). There were no statistical differences in gender composition, age, PT, APTT, INR, hemoglobin, cause of injury, compound injury, incidence of hypoxemia, incidence of high intracranial pressure, incidence of SAH, incidence of edema zone, incidence of irregular lesions, time of the first CT examination, time of the control CT examination and volume of the first CT hematoma between 2 groups ( P>0.05). The GCS, cortical distance and Fbg in progression group were significantly lower than those in non-progression group, the TT, platelet, multifocality rate and volume of the control CT hematoma were significantly higher than those in non-progression group, and there were statistical differences ( P<0.01 or <0.05). Multivariate Logistic regression analysis result showed that cortical distance <1 cm, Fbg<2 g/L and multifocality were independent risk factors affecting the progression in patients with TIPHs ( OR = 6.723, 5.515 and 4.827; P<0.05). The model had a sensitivity of 71.43% (20/28), a specificity of 92.68% (38/41), and an accuracy of 84.06% (58/69) in judging the progression of TIPHs. Conclusions:Based on the risk factors for the progression of TIPHs, predicting these patients in advance can provide necessary intervention measures for high-risk patients, which will help to reduce the rate of progression and improve the prognosis of patients.

Objective To explore the mechanism of the therapy of promoting blood circulation and nourishing yin(PBCNY)in improving spermatogenesis in rats with varicocele based on Bcl-2/Bax pathway.Methods Thirty 8-week-old male SD rats were randomly divided into six groups(five rats per group):sham,model,levocarnitine(1.0 g/kg),and PBCNY low-dose(5.2 g/kg),PBCNY medium-dose(10.4 g/kg),and PBCNY high-dose(20.8 g/kg)groups.Apart from the sham group,all the groups were built a varicocele model by the classical Turner method.After 4 weeks,the sham group and model group were gavaged with normal saline,and the remaining groups were given with levocarnitine or different doses of PBCNY decoction once a day for 4 weeks.At the end of gavage,the sperm quality of rats in each group was detected by a computer-assisted sperm analyzer,and HE staining was used to observe the testicular tissue morphology of rats in each group and performed with Johnsen score.TUNEL method was used to detect apoptosis,RT-qPCR was used to detect Bcl-2 and Bax mRNA expression,and immunohistochemical method was used to detect cytochrome C protein expression.Results Compared to the sham group,sperm total count,motility rate and percentage of sperm with grade(a+b)motility were lower in the model group of rats(P<0.05).HE staining showed disturbed arrangement of seminiferous tubules in the testicular tissue with significant changes and decreased Johnsen score(P<0.05).The result of TUNEL assay showed that the apoptosis rate was increased in the model group(P<0.05).Bax mRNA expression was increased in testicular tissue(P<0.05).The result of immunohistochemistry showed that the positive expression of cytochrome C was increased in the model group(P<0.05).Compared with the model group,the rat sperm total count in the PBCNY medium-dose group,motility rate in the PBCNY low-and medium-dose groups,and percentage of sperm with grade(a+b)motility in the PBCNY low-dose group increased(P<0.05);the pathological structure of rat testis had different degrees of improvement,and Johnsen score increased in the PBCNY medium-dose group(P<0.05);the apoptosis rate of testicular cells decreased in the levocarnitine group and all doses of PBCNY groups(P<0.05);the expression of Bcl-2 mRNA increased in the PBCNY low-dose and high-dose groups(P<0.05),and the expression of Bax mRNA decreased in the levocarnitine group and the PBCNY medium-dose group(P<0.05);the positive expression of cytochrome C decreased in the PBCNY medium-dose group(P<0.05).Conclusion The therapy of PBCNY can improve sperm quality,repair damaged testicular tissue structure and improve spermatogenic function in rats with varicocele,and its mechanism of action may be related to the activation of Bcl-2/Bax pathway and inhibition of cell apoptosis.