Objective:To establish a multidrug resistant human salivary gland adenoid cystic carcinoma cell line.Methods:Human salivary gland adenoid cystic carcinoma cell line ACC-2 was treated by 24-hour-exposure to high dose of Bleomycin(BLM)(20 ?g/ml).Drug sensitivity was evaluated by MTT assay.Cell counting was employed to make the growth curve and to calculate the cell doubling time.Flow cytometry(FCM) was used to study the cell cycle distribution and apoptosis.The colony formation ability was also observed.Results:Multidrug resistant cell line of human salivary gland adenoid cystic carcinoma was established and named ACC-2/BLM.After 10 times repeated exposure to BLM,the resistance index(RI) to BLM,5-Fluorouracil(5-FU),Cisplatin(CDDP),Cyclophosphamide(CTX),Vincristine(VCR) were 7.299,1.03,2.15,1.114 and 5.96 respectively.Compared with ACC-2,the proliferation potential of ACC-2/BLM cells was decreased.The ACC-2 apoptosis cells were much more than ACC-2/BLM cells after 9-day-treatment by BLM at 60 ?g/ml.Conclusion:ACC-2/BLM cell line has multidrug resistant characteristics.

Objective To investigate the distribution of traditional Chinese medical syndrome patterns of dengue fever, thus to standardize its clinical diagnosis and treatment and to enhance its therapeutic effect. Methods A prospective clinical trial was carried out in 210 dengue fever patients who were admitted from July to October of 2014. The clinical data of four physical examinations were collected and analyzed for the analysis of the distribution of syndrome patterns. Results ( 1) The traditional Chinese medical syndromes of 210 dengue fever patients were characterized by fever and aversion to cold, heaviness of limbs, poor appetite, headache and heaviness of head, abdominal fullness and discomfort, yellowish urine, yellowish and greasy fur, sluggish pulse. ( 2) The syndrome patterns of dengue fever were classified into blockage of damp-heat, disease involving both defensive phase and qi phase, heat attacking qi phase, heat attacking blood phase, toxicity invading pericardium, and sudden loss of yang-qi. The incidence of six patterns was in decreasing sequencing. ( 2) The results of laboratory examination showed that the decrease of white blood cell ( WBC) , neutrophil percentage and platelet count was obvious, and the haematocrit ( PLT) became disordered. The results of recheck showed taht the increase of calcitonin and C-reactive protein were not obvious, and the damage of vital organs was less. Conclusion Dengue fever can be classified into the damp-heat pestilence in traditional Chinese medical field. The syndrome patterns of dengue fever are dominated by blockage of damp-heat, and disease involving both defensive phase and qi phase, and correspondingly, the therapeutic methods should be focused on clearing heat, resolving dampness, and strengthening spleen.

Objective The effectiveness of Zhuang nationality medical lotus needle plus back cupping therapy ( Zhuang needle-cupping therapy) , Flixonase aqueous nasal spray and cetirizine tablets in treating allergic rhinitis (AR) was compared for the exploration of the therapeutic mechanism of Zhuang needle-cupping therapy. Methods A total of 200 recruited AR patients were randomly divided into four groups in the proportion of 1:1:1:1. The four groups were Zhuang needle-cupping therapy group, cetirizine group, Flixonase group and blank control group. The blank control group had no medication, and the patients of the other three medication groups were given the corresponding treatment. Ten days constituted one treatment course, and interval between two courses lasted one week. After two courses, the therapeutic effect was evaluated. The changes of specific IgE (S-IgE), leukotriene (LT), interleukin 4(IL-4), IL-9 mRNA, interferon gamma (IFN-γ), Thl / Th2 cells, and Th17 cytokine ( IL-17) were observed before and after treatment. Results ( 1) After two treatment courses, Zhuang needle-cupping therapy group had better therapeutic effect than cetirizine group , Flixonase group and blank control group, and the therapeutic effect of cetirizine group and Flixonase group was better than the blank control group (P<0.05). However, the therapeutic effect of cetirizine group was similar to that of Flixonase group ( P>0.05). ( 2) After treatment, the levels of S-IgE, LT, IL-9 mRNA, IL-4 and IL-17 were decreased, and IFN-γ and Th1/Th2 levels were increased in the three medication groups ( P<0.05 compared with those before treatment) . The differ ences of the laboratory indexes in the blank control group were insignificant before and after treatment ( P>0.05). The results of inter-group comparison after treatment showed that Zhuang needle-cupping therapy group had better effect on improving S-IgE, LT, IFN-γand Th1/Th2 than cetirizine group and Flixonase group (P<0.05). (3) During the trial, no adverse reaction was found. Conclusion Zhuang needle-cupping therapy exerts certain therapeutic effect for AR, and the mechanism may be related with the inhibition of S-IgE, LT, IL-9 mRNA and IL-17 expression, and with the regulation of Th1/Th2 imbalance by decreasing TH2 cytokine level and increasing Th1 cytokine level.

The rabies virus (RABV) is an enveloped RNA virus. It mainly damages the central nervous system and causes anencephaly in mammals and humans. There is now compelling evidence that enveloped virions released from infected cells can carry many host proteins, some of which may play an important part in viral replication. Several host proteins have been reported to be incorporated into RABV particles. However, a systematic study to reveal the proteomics of RABV particles has not been conducted. In the present study, after virus culture and purification by sucrose density gradient ultracentrifugation, a proteomics approach was used to analyze the protein composition of purified RABV particles to understand the molecular mechanisms of virus-cell interactions. Fifty host proteins, along with five virus-encoded structural proteins, were identified in purified RABV particles. These proteins could be classified into ten categories according to function: intracellular trafficking (14%), molecular chaperone (12%), cytoskeletal (24%), signal transduction (8%), transcription regulation (12%), calcium ion-binding (6%), enzyme binding (6%), metabolic process (2%), ubiquitin (2%) and other (14%). Of these, four proteins (beta-actin, p-tubulin, Cofilin, Hsc70) were validated by western blotting to be present in purified RABV particles. This novel study of the composition of host proteins in RABV particles may aid investigation of the mechanism of RABV replication.
Animals ; Humans ; Molecular Sequence Data ; Proteomics ; Rabies ; genetics ; metabolism ; virology ; Rabies virus ; chemistry ; genetics ; metabolism ; Viral Proteins ; analysis ; chemistry ; genetics ; metabolism ; Virion ; chemistry ; genetics ; metabolism

Objective To observe the curative effect and adverse reactions of intercostals nerve block combined with fentanyl transdermal system in treatment of post-thoracotomy pain syndrome (PTPS). Methods Intercostals nerve block combined with fentanyl transdermal system was used for treatment of 141 patients with PIPS. The degree of pain relief was evaluated by numerical rating scale (NRS),the quality of life of patients was evaluated by the quality of life score standard,and the adverse reactions were also observed. Results The overall remission rate was 97.85%,there were 51 cases (36.43%)with complete remission, 80 cases (57.14%)with significant remission,and 6 cases (42.85%) with moderate remission. The adverse reactions included nausea,vomiting, constipation,dizziness,skin pruritus and dysuria,no serious liver and kidney function damage and respiratory inhibition. The quality of life of all patients was improved. Conclusion Multimodal analgesia has better curative effect and less adverse reactions in treatment of PTPS,so it deserves clinical promotion.

Objective To investigate the changing pattern of antimicrobial resistance among gram-negative bacilli isolated from respiratory intensive care unit (RICU) for rational use of antimicrobial agents.Methods Antimicrobial susceptibility of 1 047 isolates of gram-negative bacilli from 2000 to 2004 was tested by disk diffusion method.WHONET 5.3 software was used to analyze the data.Results The most common pathogens were Pseudomonas aeruginosa (42.9%),Stenotrophomonas malto- philia (17.1%),Acinetobacter baurnannii (10.0%) and Klebsiella pneumoniae (6.5%).The susceptibility rate of P.aerug- inosa was relatively higher to ceftazidime (50%-74%),amikacin (33.3%-81.0%),piperacillin-tazobactam (30.4%-64.6%) and cefoperazone-sulbactam (33.5%-47.5%),while the susceptibility to imipenem decreased.The susceptibility rate of S. maltophilia was relatively higher to cefoperazone-sulbactam (47.2%-78.6%) and ticarcillin-clavulanic acid(28.3%-86.6%). More than 90% of Acinetobacter baumannii isolates were susceptible to imipenem.The susceptibility rates of K.pneumoniae to imipenem and cefepime were 92.9%-100% and 55.6%-80.0%,respectively.The susceptibility rate to piperacillin-tazobac- tam decreased from 58.3% to 21.7%.The prevalence of extended-spectrum?-lactamases (ESBLs) in K.pneumoniae increased from 11.1% in 2002 to 47.8% in 2004.Conclusions Most pathogens show significant resistance to the most commonly used an- tibiotics.It is very important to select antibiotics for the treatment of infections in ICU based on the results of susceptibility.

OBJECTIVEThere was consanguineous relationship between caspase-3 and early damage after hypoxia and ischemia. Caspase-3 plays a key role in the process of apoptosis in neuron. Magnesium sulfate could protect neuron from injuries, but the mechanism was not clear. The study was to investigate the expression of caspase-3 mRNA in the hippocampus of seven-day-old hypoxic-ischemic rats and the possible mechanism of neural protection with magnesium sulfate.

METHODSThe model of seven-day-old hypoxia-ischemia rats was established. The rats were divided randomly into 6 groups as follows: (1) normal control (n = 4); (2) sham surgery control (n = 4); (3) hypoxia-ischemia (n = 4); (4) sodium chloride injection with hypoxia-ischemia (n = 4); (5) magnesium sulfate pre-injection with hypoxia-ischemia (n = 4); (6)magnesium sulfate post-injection with hypoxia-ischemia (n = 4). The therapy groups received a bolus injection of 500 mg/kg magnesium sulfate intraperitoneally 0.5 hour before or after hypoxia-ischemia. Semi-quantitative RT-PCR was used to measure caspase-3 mRNA expression in the hippocampus 24 hours after hypoxia-ischemia.

RESULTSThe expression of caspase-3 mRNA was significantly increased in the hippocampus of the hypoxia-ischemia pups (1.88 +/- 0.36 vs 0.97 +/- 0.46, P < 0.05). The expression of caspase-3 mRNA in rats with magnesium sulfate pre-injection and post-injection decreased significantly (1.54 +/- 0.49, 1.65 +/- 0.48 vs 1.88 +/- 0.36, P < 0.05).

CONCLUSIONCaspase-3 was activated in the hippocampus of the seven-day-old rats 24 hours after hypoxia-ischemia. The suppression of the expression of caspase-3 mRNA in the hippocampus was probably related to the protective effect of magnesium sulfate on the brain injury of hypoxia-ischemia.

Animals ; Animals, Newborn ; Caspase 3 ; Caspases ; genetics ; Female ; Gene Expression Regulation, Enzymologic ; Hippocampus ; blood supply ; metabolism ; Hypoxia-Ischemia, Brain ; physiopathology ; prevention & control ; Magnesium Sulfate ; therapeutic use ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction

Abstract: Objective To investigate the epidemiological characteristics of pathogens causing bloodstream infection in hematology patients during treatment and to compare the effects of allogeneic hematopoietic stem cell transplantation (HSCT) on them, so as to provide evidence for the diagnosis and treatment of bloodstream infection. Methods A total of 292 cases with bloodstream infection in hematology wards of the PLA General Hospital were collected from 2017 to 2021, which were divided into HSCT group and N-HSCT group according to whether performed HSCT or not. The epidemiological characteristics and influence of pathogenic bacteria in blood stream infection were analyzed and compared between the two groups. Results A total of 362 strains of pathogenic bacteria were collected from 292 cases, including 106 strains in HSCT group (84 cases) and 256 strains in N-HSCT group (208 cases). Bloodstream infections were more common in acute myeloid leukemia (130/392, 44.52%), followed by non-Hodgkin's lymphoma (74/292, 25.34%). The rate of once bloodstream infection in HSCT group was higher than that in N-HSCT Group, but the rate of twice bloodstream infections in N-HSCT group was higher. Gram-negative Bacilli were the most common pathogens (56.08%), with Escherichia coli being absolutely dominant (109/362, 30.11%), followed by Klebsiella pneumoniae (39/362, 10.77%). Coagulase-negative staphylococci (CoNS) (107/362, 29.56%) were the most common Gram-positive cocci. The detection rate of fungi in HSCT group (10/106, 9.43%) was significantly higher than that in N-HSCT Group (3.52%). The drug resistance rate of the common pathogenic bacteria was at a high level, and there was a certain proportion of multi-drug resistant strains (except for Pseudomonas aeruginosa). The resistance rates of CoNS to penicillin, gentamicin, moxifloxacin, clindamycin and rifampicin in HSCT group were higher than those in N-HSCT Group. The resistance rate of Escherichia coli to piperacillin/tazobactam, cephalosporins and etapenem in HSCT group was significantly higher than that in N-HSCT group. Conclusions The pathogens of blood stream infection in hematology patients are complicated and various. It is difficult for clinical diagnosis and treatment to detect multiple infections and multiple pathogens. HSCT patients have a higher risk of fungal bloodstream infection and more multi-drug resistant strains detected. Therefore, the identification of bloodstream infection and multi-drug resistant strains associated with HSCT patients should prompt surveillance.

OBJECTIVESTo summarize hemodynamic and metabolic changes during bypass, and to evaluate the bypass in liver transplantation.

METHODSFifty-four patients underwent orthotopic liver transplantation with venovenous bypass from May 2000 to May 2002. Their clinical features were analysed.

RESULTSSHR, MAP, CVP, CO, PaO(2), PaCO(2), serum K(+), Na(+), Ca(2+), BUN values were not significantly changed during bypass. Compared to the pre-bypass stage, pH was decreased in the post-bypass stage (P < 0.05), serum lactic acid value was increased in the bypass and post-bypass stage (P < 0.05), active clotting time was increased in the bypass stage (P < 0.05), serum creatinine value was increased on first postoperative day (P < 0.05).

CONCLUSIONSVenovenous bypass could improve hemodynamic and metabolic stability in the anhepatic phase, but it also could increase operation duration, liver ischemic time and cost.

Adolescent ; Adult ; Aged ; Female ; Hemodynamics ; Humans ; Kidney ; physiopathology ; Liver Transplantation ; methods ; Male ; Middle Aged ; Portal Vein ; surgery ; Postoperative Complications ; etiology

Objective To construct the prokaryotic expression vector of human autophagy-related LC3B gene,obtain the GST-LC3B recombinant plasmid , purify the GST-LC3B fusion protein and identify its activity in vitro.Methods Human LC3B coding region was amplified from the human mammary gland cDNA by PCR and inserted into the prokaryotic expres -sion vector pGEX-KG.The recombinant plasmid pGEX-KG-LC3B was transformed into E.coli Rossate.The expressed product was purified by GST-Sepharose 4B beads and identified by SDS-PAGE and Western blot analysis .The function of the purified protein GST-LC3B was detected by GST pull-down assay.Results About 400 bp of the LC3B coding region was successfully amplified from the mammary gland library by PCR and inserted into pGEX -KG.The result of double diges-tion and sequencing showed that the GST-LC3B recombinant plasmid was successfully obtained .The GST-LC3B fusion pro-tein of about 40 000 (Mr) was successfully purified and identified by SDS-PAGE and Western blotting analysis.GST pull-down assay showed that GST-LC3B could interact with Atg4B, which identified its known function .Conclusion The pro-karyotic expression vector of GST-LC3B is constructed successfully , which will facilitate further research on the function of LC3B in autophagy.