OBJECTIVE To test and analyze the new drug-resistant characteristics of Acinetobacter baumannii.METHODS The bacteria identification was determined by the routine method.Antimicrobial susceptibility was detected by K-B test.RESULTS In 95 A.baumannii strains,in addition to imipenem,the drug-resistant incidence was above 49%.There were two A.baumannii strains resistant to imipenem,and were seen panresistant strains The new characteristics of drug-resistance were found.CONCLUSIONS A new drug-resistant mode about A.baumannii to amikacin is found.The high drug-resistant incidence of A.baumannii becomes a very important problem.

OBJECTIVE To investigate the ?-lactamases gene of multi-drug resistant Acinetobactor baumannii(MDR-ABA).METHODS The blaTEM,blaSHV,blaPER,blaGES,blaIMP,blaVIM,blaSIM,blaOXA-23,blaADC and blaDHA genes of 20 MDR-ABA strains were detected by PCR.RESULTS Among 20 MDR-ABA strains,18 strains were with positive blaADC,11 strains were with positive blaTEM;the others were negative.Totally 19 strains were with genes correlated to ?-lactam antibiotics.The results indicated that the blaADC DNA sequence was a new subtype type compared with the blaADC sequence which had registered on America GenBank.CONCLUSIONS blaADC Gene and blaTEM gene have spread in the MDR-ABA group and a new blaADC subtype has been found.

OBJECTIVE To investigae the gene resistant to quaternary ammonium compounds of multiple drug resistant Acinetobacter baumannii(MDR-ABA).METHODS The quaternary ammonium compounds qacE?1 gene in 20 MDR-ABA strains were detected by PCR.RESULTS The 20 MDR-ABA strains showed multiple resistance,its sensitivity to imipenm was 65% only.Among twenty strains of MDR-ABA qacE?1 gene were all positive.CONCLUSIONS The results indicate that qacE?1 gene were all carred class 1 integron.The chlorhexidine usage in prevnting hospital infection after surgery should be reevaluated.

OBJECTIVE To investigate the aminoglycoside modifying enzymes genes of multi-drug resistant Acinetobacter baumannii(MDR-ABA).METHODS The four kinds aminoglycoside modifying enzymes genes(aac(6′)-Ⅰad,aac(6′)-Ⅰb,aac(3)-Ⅰ,ant(3″)-Ⅰ) of 20 strains MDR-ABA were detected by PCR.RESULTS Among 20 MDR-ABA strains,12 strains of aac(3)-Ⅰ,15 strains of aac(6′)-Ⅰb,18 strains of ant(3″)-Ⅰwere positive,and aac(6′)-Ⅰad was negative.The new subtype aac(6′)-Ⅰad gene has not been found.CONCLUSIONS Twenty strains MDR-ABA aminoglycoside modifying enzymes genes are found in all the 20 MDR-ABA strains.The genotype is in accordance with antibiotics resistance.It can induce clone transmitting hospital infection.It is first time to study the aac(6′)-Ⅰad gene of A.baumannii in China.

Bone turnover is regulated by local concentrations of cytokines such as osteoprotegerin (OPG) and receptor activator of nuclear factor kappaB ligand (RANKL). To explore the in vivo biological function of Opg and the mechanism of osteoporosis due to deficiency of Opg, Opg knockout mice have been generated through homologous recombination. Opg-/- mice exhibit a sharply decrease in bone density and strength as expected. The number of osteoclasts in Opg-/- mice significantly increases. Morphologically, osteoclasts appear more cuboidal in shape in Opg-/- mice than those of wt mice, suggesting that active osteoclastogenesis occurs in the absence of Opg. In consistent with this finding, an increase of osteoblast activity was also observed with accelerated mineral accumulation rate by histomorphometric measurement and elevated serum alkaline phosphatase activity (ALP) in Opg-/- mice. Interestingly, more than 50% of 2-month-old Opg-/- mice manifest medial calcification of aorta with comparable serum concentrations of calcium and phosphorus to wt mice. In conclusion, Opg-/- mice have a high-bone-rurnover type osteoporosis. The aortic calcification in Opg-/- mice is not due to abnormality of calcium and phosphorus metabolism. The mechanism underlying aortic calcification in Opg-/- mice needs to be further investigated.

ObjectiveTo investigate the effect of the expression of HBcAg in hepatocytes on the serum level of HBcAb and seroconversion of HBeAg after antiviral therapy with nucleos(t)ide analogues (NUCs). MethodsSerum samples and liver tissue paraffin sections were collected from 101 chronic hepatitis B (CHB) patients who received antiviral therapy with NUCs in Nanfang Hospital and Panyu Central Hospital from January 2015 to June 2018. ELISA was used to measure the serum level of HBcAb, and immunohistochemistry was used to measure the expression of HBcAg in the liver. The GEO database (GSE96851) was analyzed to obtain differentially expressed genes in the liver of patients with HBcAg-positive hepatitis. The two-independent-samples t test was used for comparison of continuous data between two groups; the multiple-independent-samples nonparametric Kruskal-Wallis H test was used for comparison of continuous data between multiple groups, and Dunnett method was used for further comparisons; the chi-square test was used for comparison of categorical data between groups. ResultsThe expression pattern of HBcAg in hepatocytes was classified as absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression, and according to expression level, HBcAg expression was classified as grades Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression. HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 16.67%, 22.73%, and 24.24%, respectively, in the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression (χ2=4753, P=0.037), and HBeAg seroconversion rates after 96 weeks of antiviral therapy were 5.88%, 13.04%, 27.59%, and 26.67%, respectively, in the patients with grade Ⅰ, Ⅱ, Ⅲ, and Ⅳ expression (χ2=6.580, P=0.016). There were significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with absent expression, nuclear expression, cytoplasmic expression, and nuclear/cytoplasmic expression of HBcAg (HBcAb-IgM: H=9.760, P=0.021; total HBcAb: H=21.46, P＜0.001), and there were also significant differences in the serum levels of HBcAb-IgM and total HBcAb between the patients with grade Ⅰ, Ⅱ, Ⅲ, and IV expression of HBcAg (HBcAb-IgM: H=18.80, P＜0.001; total HBcAb: H=26.03, P＜0.001). The analysis of differentially expressed genes in the liver showed that the expression of antibody-related genes was upregulated in the liver of patients with HBcAg-positive acute liver failure. ConclusionThe expression pattern and level of HBcAg in the cytoplasm of hepatocytes are associated with serum HBcAb, and the measurement of HBcAg may help to predict the efficacy of antiviral therapy with NUCs.

Adiponectin is an adipocyte-derived secretory protein. It was found to be associated with insulin resistance, inflammation and arteriosclerosis. To further study the biological function and expression of adiponection in vivo, adipoenctin gene knock-out and LacZ gene knock-in mouse model was constructed. Gene targeting strategy was designed to replace part of exon 2 and exon 3 of adiponectin gene with full length LacZ gene in frame with remaining upstream ATG and signal peptide sequence of exon 2. The targeting vector (Adipo-LacZ-XpPNT) was constructed and verified by restriction enzyme digestion and sequencing. CJ7 ES cells were transfected with targeting vector linearized by NotⅠ digestion, selected in the medium containing both G418 and ganciclovoir. Resistant clones were screened by PCR and further confirmed by Southern blot for correct homologous recombinants. Chimera mice were obtained by routing microinjection of homologous recombined ES cells into blastocysts. After mating, mice heterozygous and further homozygous for adiponectin knockout and LacZ gene knock-in were established. Expression of both endogenous adiponectin and exogenous LacZ gene in mouse tissues and sera were detected by RT-PCR, Northern-blot, Western blot and ELISA. The results show that adiponectin was disrupted at both mRNA and protein levels. LacZ gene is expressed exclusively in adipose tissue of mutant mice. Its expression profile is identical to endogenous adiponection. Unexpectedly, LacZ activity could not be detected in both adipose tissue and serum although LacZ protein can be detected in adipose tissue but not in serum of mutant mice. In conclusion, mice homozygous for adiponectin knockout and LacZ gene knock-in have been successfully constructed. Mutant mice display LacZ expression profile identical to endogenous adiponectin albeit neither LacZ activity nor protein can be detected in serum of mutant mice.

Objective:To explore the clinical effect of ultrasound evaluation of fiberoptic bronchoscope (FB) guided tracheotomy, which can provide help for difficult tracheotomy and new operators.Methods:The operating protocol was standardized for ultrasound evaluation of FB guided tracheotomy. Ten patients with difficult tracheotomy admitted to the department of critical care medicine of Donge Hospital from October 2019 to January 2020 were enrolled. According to this protocol, FB guided tracheotomy was performed under the ultrasound evaluation, and the amount of blood loss, operation time and related complications during procedures were collected.Results:The preparation of supplies, personnel, patients and the operation, the process of FB guided tracheotomy under the ultrasound evaluation were standardized. When tracheotomy was preformed for patients with difficult tracheotomy, it was necessary to use ultrasound first to evaluate the neck condition and vascular disorientation of patients, and the tracheotomy plan (tracheotomy site, incision size, and incision depth) was designed, and then the tracheotomy process was monitored under the guidance of FB. Among the 10 patients with difficult tracheotomy, 6 were male and 4 were female; body mass index was (32.2±1.4) kg/m 2. Tracheotomy was successfully completed within 10 minutes in all the 10 patients, with less than 5 mL blood loss, and no complications occurred. Conclusion:Ultrasound evaluation of FB guided tracheotomy can improve the clinical operations and ensure patient safety.

Twenty-four endophytic actinomycetes strains were isolated from the Salvia przewalskii in Tibetan Plateau of China by tablet coating method. Fusarium moniliforme, Helminthosporium turcicum and Bipolaris maydis were selected as indicator fungi to test the antimicrobial activities of these endophytic actinomycetes by tablet confrontation method. The results showed that 21 strains can produce antimicrobial substances which accounts for 85.7% of the total separates number. Four strains of endogenous actinomyces have more obvious antifungi activity. According to results of morphology and culture properties and 16S rDNA sequences of endophytic actinomyces, it is concluded that all of the isolates were streptomycetes trains.
Actinomyces ; chemistry ; genetics ; China ; DNA, Ribosomal ; genetics ; Fusarium ; drug effects ; Helminthosporium ; drug effects ; Salvia ; microbiology

The manufacturing process of Chinese medicines is the significant link to achieve "effect-enhancing and toxicity-reducing", including an interaction between "toxicity and effect". This paper would elucidate the effects of Chinese herbal compound decoction, preparation, dosage forms, route of administration and quality of pharmaceutical excipients on "toxicity-effect" theory from the formulation approaches. The article pointed out that the comprehensive analysis on "toxicity-effect" theory should be strengthened from the aspects of overall manufacturing, fundamental research and modern Chinese preparation, to explore the mechanism of "effect-enhancing and toxicity-reducing" in the manufacturing process, clarify the core status of Chinese preparation in "toxicity-effect" theory, and ensure the security and effectiveness in traditional Chinese medicine clinical application.