AIM: To investigate the function and morphological changes of long-term cultured primary rat hepatocytes. METHODS: Rat primary hepatocytes were isolated by two-step in situ collagenase perfusion method, and then were purified by density and grade centrifugal method with Percoll. Cell viability was observed by 0.4% trypan blue. The hepatocytes were seeded into 6 wells plate with HepatoZYME-SFM medium. ALT, AST, albumin and urea levels in the supernatant were measured, CYPⅠA1 was detected with EROD method. RESULTS: (2-3)?108 cells per whole liver were obtained with viability and purity above 90% after purified with Percoll. Hepatocytes cultured in HepatoZYME-SFM grew well with normal hepatocyte morphometrics. ALT, AST levels in the supernatant decreased after 3-day culture, and kept at a stable level after 6-9 days. Albumin secretion and urea synthesis were maintained at high levels in 18 days, while CYPⅠA 1 enzyme activity was only detected in 3-6 days. CONCLUSIONS: Percoll was used to increase the viability and purity of freshly isolated rat hepatocytes. Hepatocyte morphometrics and their biological synthesized function are effectively maintained in HepatoZYME-SFM medium.

AIM: To Screen and identify human single-chain variable fragment (ScFv) specific to hepatitis B virus core protein and determine its gene sequence. METHODS: The recombinant phages were panned by HBcAg coated in a 96-pore plate and 48 clones were identified specific to HBc after three rounds of panning. The specificity of ScFv from the positive clone was determined by ELISA. Then, the soluble ScFv was expressed in E.coli. HB2151 and secreted in the supernatant. Subsequently, SDS-PAGE and dot blot were performed to identify the ScFv in the supernatant and cell lysate. The gene of ScFv specific to hepatitis B virus core protein was sequenced. RESULTS: The ScFv screened from phage antibodies has a specific combination character with hepatitis B virus core antigen. Soluble ScFv was confirmed to express in E.coli. HB2151 and secrete in the supernatant. The sequence of ScFv gene conformed to that of heavy chain and kappa chain of human immunoglubulin. CONCLUSION: Human ScFv specific to hepatitis B virus core protein has been identified by means of the phage display technology, and its gene sequence has been determined.

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.

Objective To report the distribution and antibiotic resistance of bacterial strains isolated in the First Affiliated Hospital of Anhui Medical University during 2016 for improving clinical treatment of bacterial infections. Methods Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated systems. The results were interpreted according to the clinical and laboratory standards institute (CLSI) 2015 breakpoints, and analyzed using WHONWT 5.6. Results A total of 5 406 clinical isolates were collected during 2016, of which gram positive organisms accounted for 25.6％ (1 386/5 406) and gram negative organisms 74.4％ (4 020/5 406). The strains were mainly isolated from respiratory tract (35.4％, 1 913/5 406) and urine (26.7％, 1 441/5 406). The most frequently isolated microorganisms were E. coli (23.1％, 1 247/5 406), followed by K. pneumoniae (12.1％,654/5 406). The prevalence of MRSA was 52.2％ (166/318). The prevalence of MRCNS was 80.3％ (462/575). Of the ESBLs producers, E. coli accounted for 59.7％ (745/1 247), and K. pneumoniae accounted for 30.6％(200/654). ESBLs-producing strains showed significantly higher resistance rates to most antibiotics than non-ESBLs-producing strains. All the E. faecalis strains were susceptible to vancomycin. A few (0.2％) of the E. faecium strains were resistant to vancomycin, which were all from urine specimens. A total of 1 046 (19.3％, 1 046/5 406) carbapenemresistant strains were isolated. Most Enterobacteriaceae isolates were still highly sensitive to carbapenems. Carbapenem-resistant isolates were mainly A. baumannii (41.1％, 430/1 046), P. aeruginosa (17.4％, 182/1 046)) and K. pneumoniae (12.0％, 126/1 046). Most Acinetobacter strains were resistant to most of the antibiotics tested. Conclusions Antimicrobial resistance is increasing in the clinical isolates in this hospital. We should continue to strengthen antimicrobial stewardship, prescribe antibiotics for strict indications, and improve rational antibiotic use.

Objective:To gain an in-depth understanding of the research status, hotspots, and future development trends in the field of ischemic optic neuropathy (ION).Methods:Using "ischemic optic neuropathy" as the subject heading or keyword to search for relevant literature in Chinese and English databases from January 1, 2000, to December 31, 2022. The bibliometrics method and software were applied to construct the visualization map of authors, institutions, keyword co-occurrence, outburst words, and keyword clustering.Results:A total of 1 203 ION-related articles were included, 1 106 Chinese literature and 97 English literature were included; the number of published articles in this field has fluctuated and increased in the past 20 years, mainly Chinese literature and English literature have shown a low growth trend. Chinese literature involved a total of 2 171 authors, and English literature involved 368 Chinese authors. A core team represented by Wang Runsheng, Wei Shihui, Zhong Yong, and Wei Qiping was formed among the high-yielding authors. Chinese literature involved a total of 799 research institutions, and English literature covered 119 Chinese institutions. The Xian No.1 Hospital and Beijing Tongren Hospital Affiliated to Capital Medical University respectively ranked first in the number of Chinese and English literature published in this field; 121 and 23 high-frequency keywords in Chinese and English were identified. In addition to "ischemic optic neuropathy" , compound anisodine, visual field, vision, treatment, risk factors, pathogenesis, optic nerve and rAION also appeared more frequently. The Chinese literature obtained 13 emergent words, and the English literature keywords formed 11 clusters. From the perspective of research type, the Chinese and English literature in this field mainly focued on the clinical efficacy observation of nonarteriotic Anterior ischemic optic neuropathy (NAION).Conclusions:In the past 20 years, clinical studies of ION in China have mainly focused on the treatment of NAION, risk factors, and the application of auxiliary examinations in disease diagnosis. The combination of drugs in treatment, the application of optical coherence tomography angiography, and the research on pathogenesis is still a future research trend in this field.

Objective To investigate the antimicrobial resistance in the A cinetobacter baumannii strains in different parts of China during 2012 .Methods A total of 8 739 clinical isolates of Acinetobacter were collected from 13 general hospitals and two children’s hospitals ,of which most were A . baumannii (89 .6% , 7 827/8 739 ) . Antimicrobial susceptibility testing was carried out by means of Kirby-Bauer method according to the unified protocol . The susceptibility testing data were analyzed by WHONET 5 .6 software according to CLSI 2013 breakpoints .Results Majority (85 .4% ) of the Acinetobacter strains were isolated from inpatients .The remaining 14 .6% were from outpatients and emergency room patients .Of the 7 827 strains of A .baumannii , 10 .9% ,35 .2% ,35 .7% and 43 .4% were resistant to tigecycline ,minocycline ,cefoperazone-sulbactam and amikacin , respectively .The percentage of A .baumannii resistant to imipenem and meropenem was 63 .5% and 68 .2% ,respectively . The antimicrobial resistant pattern varied in different hospitals . The resistance of A . baumannii varied between different clinical departments .A number of pandrug resistant (PDR) (20 .0% ,1 567/7 827) and multidrug-resistant (MDR) (45 .0% , 3 521/7 827 ) A . baumannii were identified . Conclusions A . baumannii is the most popular pathogenic bacteria among Acinetobacter .The antibiotic resistance of A .baumannii is still increasing .Cefoperazone-sulbactam and minocycline has good in vitro antibacterial activity against A .baumannii .The antibiotic resistance of A .baumannii varies greatly with hospital and department .

Objective To investigate the distribution and antibiotic resistance of clinical Enterobacter isolates .Methods A total of 3 031 clinical strains of Enterobacter were collected from 15 hospitals from January 1 through December 31 , 2012 . Antimicrobial susceptibility testing was performed with Kirby-Bauer or minimum inhibitory concentration method .The results were analyzed according to CLSI 2012 breakpoints .Results Enterobacter cloacae and Enterobacter aerogenes accounted for 73 .0% (2 212/3 031) and 23 .9% (725/3 031) of all the Enterobacter strains .The isolates of other Enterobacter species accounted for 3 .1% (94/3 031 ) . The main source of the isolates was respiratory tract specimen , accounting for 53 .2% (1 612/3 031) .Most (> 89% ) of the Enterobacter strains were resistant to cefazolin and cefoxitin . Generally ,54 .4% ,47 .5% and 34 .3% of the strains were resistant to cefuroxime ,cefotaxime and cefazidime ,respectively . About 6 .6% to 26 .3% of the strains were resistant to amikacin ,gentamicin ,piperacillin-tazobactam ,cefepime ,cefoperazone-sulbactam ,ciprofloxacin and trimethoprim-sulfamethoxazole .Imipenem ,meropenem and ertapenem showed the highest activity , to which only 3 .5% ,3 .7% and 10 .3% of the strains were resistant ,respectively .About 8 .9% (269/3 031) of the strains were resistant to at least imipenem ,meropenem or ertapenem .Four Enterobacter strains were extensive-drug resistant (XDR) .Conclusions The prevalence of antibiotic resistance in Enterobacter isolates decreased slightly in 2012 compared to the data in 2011 ,but the situation is still very serious .We should continue to take effective measures to control the resistant strains .

Objective To investigate the distribution and susceptibility of carbapenem-resistant Enterobacteriaceae (CRE) isolates in 2012 from CHINET surveillance .Methods Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated Systems .Results were analyzed according to the breakpoints of CLSI 2012 M100-S22 .Results A total of 1 499 CRE isolates were collected from January to December 2012 ,of which K lebsiella spp .,Enterobacter spp .and E .coli accounted for 63 .5% ,15 .1% and 13 .7% ,respectively .Of the 1 499 isolates , 48 .2% and 29 .3% were from respiratory tract and ICU , respectively . The results of antimicrobial susceptibility testing showed that the resistance rate of CRE isolates to most antimicrobial agents was 70 .0%-100% except amikacin (46 .9% ) and trimethoprim-sulfamethoxazole (49 .8% ) .CRE isolates from adults were more resistant to ciprofloxacin ,aminoglycosides and trimethoprim-sulfamethoxazole than those from children .Conclusions The antibiotic resistance of CRE isolates is very high . The spread of CRE strains in a specific region such as ICU and neurosurgery ward poses a serious threat to clinical practice and implies the importance of strengthening infection control .

Objective To investigate the susceptibility and resistance of clinical isolates collected from hospitals in several regions of China . Methods Fourteen general hospitals and two children ’ s hospitals were involved in this program . Antimicrobial susceptibility testing was carried out according to a unified protocol using Kirby-Bauer method or automated Systems .Results were analyzed according to CLSI 2013 breakpoints .Results A total of 84 572 clinical isolates were collected from January to December 2013 ,of which gram negative organisms and gram positive cocci accounted for 73 .0% and 27 .0%respectively .Methicillin-resistant strains in S .aureus (MRSA) and coagulase negative Staphylococcus (MRCNS) accounted for an average of 45 .2% and 73 .5% respectively .The resistance rates of methicillin-resistant strains to β-lactams and other antimicrobial agents were much higher than those of methicillin-susceptible strains .However ,92 .2% of MRSA strains were still susceptible to trimethoprim-sulfamethoxazole while 87 .4% of MRCNS strains were susceptible to rifampin . No staphylococcal strains were found resistant to vancomycin ,teicoplanin or linezolid .In Enterococcus spp .,the resistance rates of E . f aecalis strains to most tested drugs (except chloramphenicol) were much lower than those of E . f aecium .Some strains of both species were resistant to vancomycin .Vancomycin-resistant strains of E . f aecalis and E . f aecium were mainly VanA type based on their phenotype .Regarding non-meningitis S . pneumoniae strains ,the prevalence of penicillin-susceptible S . pneumoniae and penicillin-intermediate S . pneumoniae strains isolated from both adults and children were lower than those isolated in 2012 ,but the prevalence of penicillin-resistant S .pneumoniae strains increased .The prevalence of ESBLs producing strains was 54 .0% in E .coli ,31 .8% in Klebsiella spp .(K .pneumoniae and K .oxytoca) and 16 .5% in Proteus mirabilis isolates on average . ESBLs-producing Enterobacteriaceae strains were more resistant than non-ESBLs-producing strains in terms of antibiotic resistance rates .The strains of Enterobacteriaceae were still highly susceptible to carbapenems .Overall less than 7 .0% of these strains were resistant to carbapenems .About 62 .8% and 59 .4% of Acinetobacter spp .(A .baumannii accounts for 89 .2% ) strains were resistant to imipenem and meropenem ,respectively .Compared with the data of year 2012 , extensively-drug resistant strains in K . pneumoniae and A . baumannii decreased .Conclusions The antibiotic resistance of clinical bacterial isolates is growing in 2013 .The disseminated multi-drug or pan-drug resistant strains in a special region poses a serious threat to clinical practice and implies the importance of strengthening infection control .