Objective To observe the effect of rutin on growth and proliferation of human hepatic cancer line(HepG2).Methods HepG2 cells were cultured in vitro,then cocultured with 50 to 250 ?mol/L rutin for 24 h.The inhibition rate of rutin on growth and proliferation of HepG2 was determined by MTT,~(3)H-TdR,and apoptotic cells were observed in fluorescent staining by Olympus fluorescent microscopy,and cell cycle was analysed by flow cytometry.Results Rutin inhibited HepG2 cells from growth and proliferation,and evoked apoptosis.Flow cytometry showed that 50 to 250 ?mol/L rutin caused an increase at G_(0)/G_(1) phase and a decrease at G_(2)/M phase and arrest at G_(0)/G_(1) phase in the cell cycle.Conclusion Rutin markedly inhibits the proliferation of HepG2 cells and induces apoptosis in a concentration-dependent manner.

Objective Finding out the kinds and activities in protease of brain,heart,lung,kidney,eyes,skin and muscle of Andrias davidianus. Methods Using protease returned electrophoresis technique(G-PAGE). Results 1.Activity of proteases in brain was weak at pH 4.5,and showed no activity at pH 7.0 and pH 9.5;2.Proteases of heart and lung had activities at pH 4.5 and pH 7.0;3.Activity of kidney proteases was strong at pH 4.5 and stronger at pH 7.0,and kinds of proteases were more at pH 7.0 than at pH 4.5;4.Proteases in eyes had no activity at pH 4.5 and pH 9.5;5.Activity of proteases in muscle was very strong at pH 7.0 and pH 9.5,and kinds of proteases were more at pH 7.0 than at pH 9.5;6.Activity of skin proteases was similar to that of muscle,but the former was weaker than the later. Conclusions Among the seven kinds of tissues and organs of Andrias davidianus,kinds and activities of kidney and muscle proteases were stronger and more than others;and the eyes showed almost no protease activity.The proteases-activity optimal pH of brain was acidic;The proteases-activity optimal pH of heart,lung and kidney was acid of neutral;and the proteases-activity optimal pH of skin and muscle was neutral.

Objective To investigate the effects of spleen preservation on hepatic fibrosis and relevant cytokine in rabbits with advanced schistosomiasis. Methods After hepatic cirrhosis was induced by infecting Schistosoma japonicum cercariae in rabbits, total splenectomy (TSG), subtotal splenectomy (SSG) or sham operation (model control group, MCG) were performed respectively on these rabbits. Meanwhile,a normal control group (NCG) was established. The serum levels of tumor necrosis factor alpha (TNF-α) , interleukin-6 (IL-6) and interleukin-1 beta (IL-lβ) were detected respectively by radioimmunoassay(RIA) at the 8th, 15th and 21st week post-infection. The expressions of transforming growth factor betal (TGF-β1), type Ⅰ and type Ⅲ collagen in liver tissues were determined by immunohistochemistry before and after the operations. Results Compared with NCG, the serum levels of TNF-α, IL-6 and IL-1β of MCG rabbits increased significantly at the 8th week post-infection (P <0.01). However, the levels of them decreased to a lower level at the 15th week. At the 6th week after operation,no significant difference was found among the three model groups ( MCG, TSG, SSG) (P > 0.05). The expressions of TGF-β1, type Ⅰ and type Ⅲ collagen in liver tissue of MCG rabbits were significantly higher than those of NCG rabbits before the operation (P < 0. 01). No significant difference was found among the three model groups at the 6th week after the operation ( P > 0.05). Conclusion The residual splenic tissue after subtotal splenectomy does not aggravate the hepatic fibrosis at advanced schistosomiasis. The mechanism may be that the relevant cytokines of hepatic fibrosis (TGF-β1, TNF-a, IL-6, IL-1β) decreased to a lower level at this time,and splenectomy does not influence the levels of them.

Objective To investigate the incidence and causes of complication of stenting in the treatment of superior vena cava syndrome (SVCS) for its proper prevetions. Methods Thirty nine patients with SVCS due to various diseases were retrospectively analyzed. The location, extension and degree of the stenosis were identified on venography before stent placement. Lumen narrowing becoming less than 50% or 20% improvement after stenting was taken as effective. Complications after stenting were analyzed. Results Thirty-five(89.7%, 35/39) patients relieved after stenting. Six patients (15.4%) had complications, including stents displacement due to mass lessened after chemotherapy in 1 cases, stents displacement and thrombogenesis due to lack of anticoagulant therapy and mass lessened after chemotherapy in 1 cases, restenosis due to mass growing into the screening of Gianturco Z-type stent in 1 cases, acute pericardial effusion in 1 cases, 1 of acute pulmonary embolism(PE) in 1 cases and acute congestive right heart failure and relieved after treatment in 1 cases. Conclusions Some complications of stenting in SVCS is serious. Proper procedure, stents selection and skillful handling can reduce the frequency of the complications.

Objective To explore the bacteriostatic efficacy of snake venom antibacterial peptide OH-CATH and chitosan as ar-tificial implant coating on escherichia coli (E .coli) .Methods The catheters and dacron patches (blank group ,chitosan group ,cef-operazone group and antibacterial peptide group) were performed the pretreatment .Then the in vitro bacteriostatic test was conduc-ted to evaluate the antibacterial activity U value on E .coli ATCC 25922 and cephalosporin-resistant E .coli on 1 d and on 7 d after putting it into plasma .Results The cefoperazone group and the antibiotic peptide group had the very powerful antibacterial activity on E .coli ATCC 25922 on 1 ,7 d ,but the antibacterial activity of the cefoperazone group was stronger than that of the antibiotic peptide group(P<0 .05);the cefoperazone group had no antibacterial activity on cephalosporin-resistant E .coli ,but the antibiotic peptide group had significantly antibacterial activity on 1 ,7 d(P<0 .01) .Conclusion Snake venom antibacterial peptide OH-CATH has very obvious antibacterial effect on the standard strain and drug-resistant strain of E .coli and its value applied in artificial im-plant coating is affirmed .

Photoautotrophic cultivation with heterotrophic cells as seeds (heterotrophic cells/photoautotrophic cultivation) is an effective way for the development of microalgal biofuel, but its development potential from the point of process optimization has not been investigated in literatures. To evaluate this, the optimizations of medium and culture conditions for Chlorella ellipsoidea were studied. In the heterotrophic stage, the biomass concentration reached 11.04 g/L with the optimized medium in flask, which were 28.0% higher than that with the original medium, and the biomass concentration reached 73.89 g/L in 5-L fermenter. In the photoautotrophic stage, the culture medium and conditions were studied in a 2-L column photobioreactor. The maximum biomass concentration, lipid content and lipid productivity reached 1.62 g/L, 36.34% and 6.1 mg/(L·h) under the optimal photoautotrophic conditions. The lipids were mainly composed of C16-C18 fatty acids, which were raw material suitable for biodiesel. After optimization, heterotrophic cells/photoautotrophic cultivation can significantly improve the capacity of biofuel production by Chlorella ellipsoidea, this method is also expected to be an efficient way for the cultivation of other microalgae that can grow heterotrophically.
Biofuels ; Biomass ; Cell Culture Techniques ; Chlorella ; metabolism ; Culture Media ; Fatty Acids ; biosynthesis ; Heterotrophic Processes ; Lipids ; biosynthesis ; Photobioreactors

The HBV X gene was amplified by PCR according to the pecob6 containing the whole fragment of adw subtype of HBV, then the fragment was inserted into the multiple clone site of pAdTrack-CMV. The linearized shuttle plasmid was homogenously recombined with AdEasy-1 in BJ5183 and the recombinant adenoviral plasmid pAd-X was generated. Then plasmid pAd-X was digested with Pac I and transfected into 293 cells for packaging and amplifying. Infection titer and rate were monitored by green fluorescent protein (GFP) expression. With restriction endonuclease analysis and PCR methods, it has been confirmed that HBV X gene was cloned into the adenovirus vector successfully. The expression of X protein in HepG2 cells was detected by Western-blot. The recombined adenovirus Ad-X was constructed successfully, which would contribute to the advanced functional study of HBV X protein.
Adenoviridae ; genetics ; metabolism ; Genetic Vectors ; genetics ; metabolism ; Green Fluorescent Proteins ; metabolism ; Hep G2 Cells ; Humans ; Recombinant Proteins ; biosynthesis ; genetics ; Trans-Activators ; biosynthesis ; genetics ; Transfection

We wished to undertake molecular genetic typing and evaluate recombinants of the hepatitis-B virus (HBV) in Tibet (China). Multistage random sampling was used to collect HBsAg-positive samples. Nested polymerase chain reactions were used to amplify the whole sequence of the HBV. DNAstar, MEGA6 and SimPlot were used to assemble sequences, create phylogenetic trees, and undertake recombination analyses. Twelve whole sequences of the HBV of a Tibetan population were collected using these methods. Results showed that all 12 strains were C/D recombinants. Nine of the recombinations were at nt750, and the other three at nt1526. Therefore, the 12 strains could be divided into two types of recombinants: C/Da and C/Db. Analyses of the sequence of the whole genome revealed that the 12 strains belonged to genotype C, and that the nucleotide distance was > 4% between the 12 strains and sub-genotypes C1 to C15 in Genbank. The most likely sub-genotype was C1. Individuals with C/Da were from central and northern Tibet (e.g., Lasa, Linzhi, Ali) and those with C/Db recombinants were from Shannan in southern Tibet. These data suggest that the two types of recombinants had a good distribution in Tibet. Also, they can provide important information for studies on HBV recombination, gene features, virus evolution, as well as the control and prevention of HBV infection in Tibet.
Adult ; Female ; Genotype ; Hepatitis B ; virology ; Hepatitis B virus ; classification ; genetics ; isolation & purification ; Humans ; Male ; Phylogeny ; Recombination, Genetic ; Sequence Analysis, DNA ; Tibet ; Young Adult

Objective:To analyze the risk factors of recurrent wheezing in children with bronchiolitis and to construct a predictive model.Methods:Prospective research methods was used. One hundred and eighty children with bronchiolitis who were treated in Hefei Eighth People's Hospital from February 2017 to February 2019 were selected as the study subjects, and the included children were separated into a modeling group (126 cases) and a validation group (54 cases) according to 7∶3. The children were followed up for 3 years, and then the modeling group was divided into wheezing group (48 cases) and no wheezing group (78 cases) according to whether the children had recurrent wheezing. The Hosmer-Lemeshow fitting curve and receiver operating characteristic (ROC) curve were drawn to evaluate the validity and accuracy of the constructed prediction model.Results:Multivariate Logistic regression analysis showed that artificial feeding ( OR = 8.838, 95% CI 2.601 to 30.027), family history of allergies ( OR = 6.709, 95% CI 1.825 to 24.665), underlying diseases ( OR = 8.114, 95% CI 1.638 to 40.184), and higher IgE level ( OR = 1.020, 95% CI 1.012 to 1.029) were the independent risk factors for recurrent wheezing in children with bronchiolitis ( P<0.05). The area under the curve of the modeling group was 0.917 (95% CI 0.855 to 0.959), and the sensitivity and specificity were 83.33% and 85.90%, respectively; the area under the curve of the validation group was 0.911 (95% CI 0.847 to 0.954), and the sensitivity and specificity were 89.58% and 79.49%, respectively. Conclusions:Artificial feeding, family history of allergies, underlying diseases, and higher IgE level are the independent risk factors for recurrent wheezing in children with bronchiolitis. The constructed prediction model has good accuracy and validity, and can be used as an effective tool for clinical prediction of recurrent wheezing in children with bronchiolitis.

Objective To summarize the clinical features of 9 cases with mutations in PKHD1 gene for a better understanding of its phenotype.Methods Clinical data of nine cases with mutations in PKHD1 gene were summarized from January 2011 to December 2016 in our center,including clinical manifestations,laboratory findings,imaging data and family investigation.Next generation sequencing was used to screen 4000 genes in case 1 to 4 and whole exons in case 5 to 9.Significant variants detected by next generation sequencing were confirmed by conventional Sanger sequencing.Segregation analysis was performed using parental DNA samples.Relevant literature was reviewed.Results Among these 9 cases,5 are male,4 are female.The average age of onset was 2.6 years old (ranging from 0.5-5.2 years).Renal ultrasound revealed that all 9 cases had cysts in bilateral kidney,7 cases with enlarged kidney,1 case with normal size kidney,1 case with normal size kidney,and 1 case with bilateral renal atrophy.Two cases with renal artery stenosis,1 case with focal narrowing in left main branch and 1 case with vesico-ureteral reflux were found.Among the 9 cases,3 cases had homozygous mutations,and 6 cases had compound heterozygous mutations,including 1 nonsense mutation,1 frameshift mutation and 15 missense mutations.There were 2 cases with 3 heterozygous mutations,2 c.5935C ＞ T mutations and 2 eases with C.5869G ＞ A mutations.A total of 10 new mutations were identified.Conclusion Patients with mutations in the PKHD1 gene had normal size kidney,or even atrophic kidney.Renal artery stenosis,vesicoureteral reflux and bronchial stenosis were all first reported in patients with mutations in PKHD1 gene.The novel mutations,c.274C ＞ T,c.9059T ＞ C,c.8996delG,c.281C ＞ T,c.10424T ＞ A,c.7092T ＞ G,c.4949T ＞ C,c.5869G ＞ A,c.6197A ＞ G and c.1877A ＞ G further expanded the mutation spectrum of PKHD1 gene.