OBJECTIVE:To study the difference between the hydrophilic and hydrophobic fluoroquinolones(FQNLs)in?ducing resistant Klebsiella pneumoniae(Kpn)in vitro.METHODS:Hydrophilic ciprofloxacin(CIP)and hydrophobic sparfloxacin(SPFX)were used to induce FQNLs resistant mutants from a clinical susceptible isolate Kpn909with stepwise selection method,respectively,The susceptivity of bacteria to was determined by the standard method of two fold agar dilu?tion.RESULTS:The resistance of Kpn909to FQNLs was raised up gradually as FQNLs'concentration increased;Minimum inhibitory concentrations(MICs)of Kpn9098Selected by CIP at2?MIC(0.5mg/L)were16～32times higher than that of Kpn909,and showed resistant to ofloxacin(OFX),lomefloxacin(LMFX)and enofloxacin(ENX)with a MIC of8mg/L,16mg/L and8mg/L.Otherwise,SPFX-selected Kpn with a similar resistant level occurred at the concentration of8?MIC(1mg/L).The maximal selecting concentration of CIP and SPFX were128mg/L and64mg/L,and MIC of5FQNLs to the resistant mutants obtained at which were≥256mg/L and32～128mg/L,respectively.CONCLUSION:Both hydrophilic CIP and hydrophobic SPFX could induce Kpn the cross resistance to others FQNLs,but hydrophobic FQNLs may be superior to hydrophilic FQNLs to delay the resistance occurrence in Kpn.

Objective To survey the antimicrobial resistance of nosocomial Klebsiella Pneumoniae (Kpn) isolates. Methods All the 68 Kpn strains were isolated and collected from nosocomial infections during April 2002 to March 2003. The susceptivity of bacteria and the production of extended spectrum ?-lactamases (ESBLs) were determined by Kirby-Bauer disc diffusion test and phenotypic confirmatory testing, respectively. Results All 68 Kpn strains were 100% susceptible to Menopenem and Imipenem but resistant to 14 other antimicrobials to different degrees. The first five drugs of high resistant rate were ampicillin (100%), amoxycillin/clavulanic acid (79.4%), sulphamethoxazole/trimothoprim (67.6%), cefotaxime (58.8%), gentamycin (47.0%). There were 28 isolates producing ESBLs (41.2%), which showed much higher resistance to ?-lactam, aminoglycosides, quinolones and sulphamethoxazole/trimothoprim than the non-ESBLs-producing strains. Conclusion Nosocomial Kpn isolates were most multidrug resistant, of which ESBLs-producing strains showed a higher resistance and which may involve other resistant mechanisms.

Objective To explore the differential liver plasma membrane (PM) proteins that may be related to the occurrence,development and reversal process of hepatitis and to understand the pathogenesis of hepatitis and the new drug targets by performing a comparative proteomics research of liver PM between hepatitis B surface antigen (HBsAg) transgenic mice and wild-type C57 mice.Methods A 6-month-old HBsAg transgenic mouse model was established.The pathological examination was performed to observe the pathological changes of transgenic mice and wild-type C57 mice.The PM from liver tissue of 6-month-old transgenic mouse and the control mouse were purified through twice sucrose density grade centrifugation combined with second antibody magnetic bead enrichment.The purity of extracted PM was verified by Western blot.Differential proteome expression analysis was performed by using two-dimensional electrophoresis (2-DE) and ImageMaster software analysis.The differentially expressed proteins were lysed by trypsin and identified by liquid chromatography combined with tandem mass spectrometry (LC-MS/MS) analysis.Results The pathological examination results showed that hepatitis was observed in the transgenic mouse group,while no abnormity was found in the controls.The PM was successfully enriched and the mitochondria contamination was reduced by sucrose density grade centrifugation combined with second antibody magnetic bead purification treatment.Thirty differential mice liver PM protein spots were visualized,in which 11 non-redundant proteins were successfully identified by LC-MS/MS in transgenic mouse group,including 9 up regulated protein spots and 2 down-regulated protein spots.These differentially expressed proteins included keratin,cardiac Ca2+ release channel,cytochrome B5,ATP synthase subunit alpha,etc.Conclusions A batch of HBsAg gene expression related differential proteins are identified in mouse liver plasma.These proteins might be new drug targets for anti-HBV treatment.This study will guide further investigation on the mechanism of HBV infection induced hepatitis.

Objective Conventional examination for vision function adopts subjective psychophysics methods.Pattern visual evoked potential (P-VEP) binocular summation response is a new objective way for the test of vision function.But its clinical value in evaluating vision function of amblyopia children is still in controversy.This study was to explore the binocular vision and the feature of P-VEP binocular summation in children with amblyopia and evaluate the significance of P-VEP binocular summation in binocular vision.MethodsThis is a case-controlled study.P-VEP binocular summation response and single ocular P-VEP response was respectively recorded in 151 hyperopia amblyopia children and 80 age- and gender-matched normal children.P-VEP response from children with hyperopia amblyopia was recorded under the corrected vision.The hyperopia amblyopia was diagnosed based on the standard of National Children Amblyopia and Strabismus Prevention and Treatment Working Group.The feature of P-VEP binocular summation was further analyzed and compared with monocular VEP response.Informed consent was obtained from each subject or custodian prior to the trail.ResultsNo significant difference was found in latency between P-VEP binocular summation response and monocular P-VEP response whether amblyopia children or normal children(P＞0.05).Binocular response/monocular response value in amblyopia children was lower than that in normal children(P＜0.05).Binocular response/monocular response value in mild or moderate amblyopia children was higher than that in severe amblyopia children(P＜0.05).Binocular response/monocular response value was significantly reduced in the children with binocular vision than in the children without binocular vision (P＜0.05).ConclusionP-VEP binocular vision,acting as an evaluation factor,could impartially reflect the function of binocular vision of amblyopia children.

Objective:To investigate therapeutic efficacy and mechanisms of action of oncolytic agent derived from herpes simplex virus type 2 (oHSV2) in a xenograft mouse model bearing CT26 colorectal cancer. Methods:BALB/c mice were subcutaneously inoculated with CT26 cells to establish a xenograft mouse model of colorectal cancer. 1) After intratumoral administration of oHSV2, enzyme-linked im-munosorbent assay was used to determine granulocyte-macrophage colony-stimulating factor (GM-CSF) expression levels in the blood. 2) Model mice were divided into three groups:PBS group (negative control), oHSV2 group, and 5-fluorouracil (5-FU) group (positive control). After drug administration, drug effectiveness was evaluated on the basis of weight, tumor volume, general state, and survival time. 3) Cells from the draining lymph nodes (TDLN) and tumor were surgical y removed and used to quantify mature dendritic cel s (DCs) and T lym-phocytes by flow cytometry. Result:1) In the CT26 xenograft model, level of GM-CSF continuously elevated. At day 8, peak value was attained in the blood at concentration of 3150±327.1 pg/mL. Then, GM-CSF expression gradually reduced as time progressed. 2) In in vivo study, both oHSV2 and 5-FU exerted antitumor effects relative to PBS group (50 days vs. 36 days, P<0.01;51 days vs. 36 days, P<0.01), and oHSV2 proved to be less toxic and safer. At day 28, the 5-FU group presented highly significant difference in mouse body weight compared with that of PBS group (16.61 g vs. 22.07 g, P<0.01). However, oHSV2 group did not show statistical y significant change (al P>0.05). Skin of virus injection region did not present necrosis and ulceration. 3) In the TDLN, the frequency of DC was increased when treated with oHSV2 compared with the control group (6.49%vs. 3.73%, P<0.01). Similarly, the percentage of CD4+and CD8+T-cel s from the oHSV2-treated group was signifcantly higher than mock-treated tumors (15%vs. 8.57%, P<0.01;8.19%vs. 5.15%, P<0.01). However, number of cells in the 5-FU group were significantly reduced with respect to that of the negative group (al P<0.01). Conclusion:oHSV2 exerted potent antitumor effects in a murine colorectal cancer model. Compared with 5-FU, oHSV2 treatment caused fewer side effects. Such antitumor effect may be induced by stimulation of immune activity by GM-CSF production.

Objective: To investigate protective effects of nicotinamide mononucleotide (NMN) on ethanol-induced DNA damage in L02 cells, so as to provide the evidence for adjuvant therapy of NMN on alcoholic liver diseases. Methods: L02 cells were pretreated with different concentrations of NMN (0, 1, 2, 4 and 8 mmol/L) for 6 h, and then were exposed to 0.4% ethanol for 12 h. The treated cells were divided into the control group, 0.4% ethanol group and different concentrations of NMN groups. Cell viability was analyzed using trypan blue staining for determining the concentration of NMN as a protective agent. The effects of NMN on ethanol-induced DNA damage in L02 cells were evaluated using immunofluorescence detection and reactive oxygen species (ROS) assay. L02 cells were exposed to 0.4% ethanol for 12 h, cultured in a medium containing a protective concentration of NMN, and divided into PBS group and NMN group. Cell viability was detected at 0, 2, 4, 8, 16 and 32 h, and the effects of NMN on repairing ethanol-induced DNA damage were evaluated by alkaline comet assay. Results: The cell viability was lower in 0.4% ethanol group than than in the control group, and was higher in different concentrations of NMN groups than in 0.4% ethanol group (all P<0.05), with no significant difference in the cells viability between 4 mmol/L and higher concentrations of NMN groups and the control group (all P>0.05). Therefore, 4 mmol/L NMN was selected as a protective agent. The cell tail moments, relative immunofluorescence intensities of γH2AX and relative levels of ROS were higher in 0.4% ethanol group than in the control group, and lower in 4 mmol/L and higher concentrations of NMN groups than in 0.4% ethanol group (all P<0.05). The cell viability was increased and the cell tail moment was shortened with the increase of 4 mmol/L NMN intervention time; and the cell viability in 4 h and more of NMN groups were higher, and the cell tail moment were lower than that in PBS group (all P<0.05). Conclusions NMN attenuates DNA damage in a dose-dependent manner and promotes the repair of DNA damage in a time-dependent manner. NMN has a protective effect on ethanol-induced DNA damage in hepatocytes.

Objective To explore the functional orientation of township hospitals in developed areas to achieve goals of the ongoing health reform.Methods Two rounds of Delphi consultation was made among 18 experts from around the country to learn the degree of importance of every function.Results The experts' average seniority of service in their fields was 23.28±10.41 years.The positivity coefficients of the two-round consultation were 100％.Kendall' S coefficients of the two-round consultation were 0.226 and 0.265 (p＜0.01).These data are of statistical importance,suggesting consensus among these experts and the consultation meeting the needs of forecast.In these consultation,seven class-1 functions and and 27 class-2 functions were determined.Medical services and public health services account for similar proportion of work of township hospitals.Conclusion The functional orientation was feasible and forward looking,for reference of township hospitals in developed areas.

Objective:To investigate the application value of three-dimensional (3D) printing technology assisted laparoscopic anatomic liver resection of segment 8 (Lap-S8).Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 8 liver cancer patients including 7 cases with hepatocellular carcinoma and 1 case with intrahepatic cholangio-carcinoma who underwent 3D printing technology assisted Lap-S8 in the Hunan Provincial People′s Hospital from January 2019 to December 2020 were collected. There were 7 males and 1 female, aged from 49.0 to 80.0 years, with a median age of 56.5 years. Of the 8 patients, 6 cases underwent laparoscopic anatomic liver resection of the entire segment 8, 1 case underwent laparoscopic anatomic liver resection of ventral subsegmental of the segment 8 and 1 case underwent laparoscopic anatomic liver resection of dorsal subsegmental of the segment 8. 3D printing technology was used to assist preoperative evaluation and intraoperative navigation for all 8 patients. Observation indicators: (1) surgical situations; (2) postoperative situations; (3) follow-up. Follow-up was conducted using outpatient examination, internet or telephone interview to detect survival and tumor recurrence of patients after operation up to March 2021. Measurement data with normal distribution were represented as Mean±SD, and measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers. Results:(1) Surgical situations: all the 8 patients underwent 3D printing technology assisted Lap-S8 successfully, without conversion to open surgery. The operation time, hepatic portal occlusion time and volume of intraoperative blood loss of the 8 patients were (216±41)minutes, (56±11)minutes and 75 mL(range, 50 to 300 mL), respectively. There was no intraoperative blood transfusion in 8 patients, and the surgical margin of the 8 patients was negative. (2) Postoperative situations: the duration of postoperative hospital stay of the 8 patients were (9±3)days. There was no complication such as postoperative hemorrhage, biliary fistula, liver abscess or abdominal infection occurred. (3) Follow-up: all the 8 patients were followed up for 3.0?24.0 months, with a median follow-up time of 12.5 months. During the follow-up, 1 of 8 patients with preoperative diagnosis of recurrent hepatocellular carcinoma developed tumor recurrence at 5 months after operation. The patient underwent laparoscopic surgery followed with the transcatheter arterial chemoembolization and target therapy, and survived with tumor. There was no tumor recurrence in the other 7 patients.Conclusion:3D printing technology assisted Lap-S8 is safe and feasible.

Objective To determine the median effective plasma concentration (Cp50) of remifentanil inhibiting cardiovascular response to CO2 pneumoperitoneum stimulus when combined with propofol in patients undergoing laparoscopic gynaecological surgery.Methods Twenty-two female patients scheduled for elective laparoscopic gynaecological surgery under general anesthesia, aged20-60 years, with a BMI 18-30 kg/m2, falling into ASA physical statusⅠ orⅡ, were enrolled in this study.Anesthesia was induced with propofol and remifentanil target-controlled infusion and iv injection of rocuronium 0.6 mg/kg.The target plasma concentration (Cp) of remifentanil and propofol were set at 5 ng/ml and 4μg/ml respectively.3 minutes after endotracheal intubation, the Cp of remifentanil was adjusted.The target Cp was set at 6 ng/ml in the first patient.CO2 pneumoperitoneum was performed after the target effect-site concentration and Cp were balanced.Each time Cp increased/decreased by 20％in the next patient depending on whether or not the cardiovascular response to CO2 pneumoperitoneum occurred.The positive cardiovascular response was defined as HR and/or MAP increasing by 20％ within 3 minutes after CO2 pneumoperitoneum.The Cp50 and 95％ confidence interval (CI) of remifentanil required to inhibit cardiovascular response to CO2 pneumoperitoneum stimulus when combined with propofol in patients undergoing laparoscopic gynaecological surgery were calculated.Results The Cp50 (95％ CI) of remifentanil required to inhibit cardiovascular response to CO2 pneumoperitoneum stimulus was 4.58 (4.14-5.08) ng/ml when combined with propofol in patients undergoing laparoscopic gynaecological surgery.Conclusion The Cp50 of remifentanil required to inhibit cardiovascular response to CO2 pneumoperitoneum stimulus was 4.58 ng/ml when combined with propofol in patients undergoing laparoscopic gynaecological surgery.

Nowadays,surgery is a comprehensive approach for the treatment of hepatic carcinoma as the first choice,but there are still many limitations in surgical therapy,such as the location,boundary and metastasis of hepatic carcinoma.Currently,the indocyanine green fluorescence imaging-guided hepatectomy is widely used at home and abroad as a new progress and hotspot in hepatobiliary surgery,which makes hepatobiliary surgery more convenient and makes up for some deficiencies in hepatectomy.Thus,we summarized the experience of indocyanine green fluorescence imaging-guided hepatectomy in the People's Hospital of Hunan Province and discussed its value of application.