OBJECTIVE To study the drug-resistant diversity of Gram-negative bacilli isolated from inpatients during recent five years.METHODS A total of 1 464 Gram-negative bacilli isolated were detected and retrospectively analyzed from 1999 to 2003.Antimicrobial susceptibility testing was performed using Kirby-Bauer method.RESULTS The resistance of Pseudomonas aeruginosa to piperacillin rised from 17.6% of 1999 to 79.2% of 2003,and that to ciprofloxacin rised from 4.3% of 1999 to 36.0% of 2003.The resistance of Escherichia coli to quinolones was above 50%,while to third-generation cephalosporins was 30-40%;the resistance of E.coli to piperacillin rised from 42.9% of 1999 to 68.9% of 2003,and that to ciprofloxacin rised from 40.0% of 1999 to 73.5% of 2003.The resistance of Acinetobacter to piperacillin rised from 31.2% of 1999 to 67.5% of 2003,and that to ceftriaxone rised from 36.0% of 1999 to 74.1% of 2003.The resistance of Serratia to ceftazidime,ceftriaxone,gentamicin,amikacin and piperacillin rised sharply.Imipenem was the most active antibiotic tested against Gram-negative bacilli.Cefoperazone/sulbactam and piperacillin/tazobactam also showed excellent activity against Gram-negative bacilli.CONCLUSIONS During recent five years,the resistance of the most common Gram-negative bacilli has increased rapidly.How to delay the resistance development of common strains become a global problem.

Response Surface Methodology was applied to optimize the culture components for lipopeptide production by Bacillus natto TK-1. In the first step, two level factorial design of Plackett-Burman was used to evaluate the influence of six related factors. It showed that three factors playing the important roles in the medium, including peptone, yeast extract powder and CaCl_2. The path of steepest ascent was used to approach the optimal region of the fermentation conditions subsequently. In the third step, the concentrations of those three main factors were further optimized by using Box-Behnken and Response Surface Analysis. By solving the quadratic regression model equation, the optimal concentrations of the variables were determined as: peptone 1.73%, yeast extract powder 0.063 %, CaCl_2 1.385?10-4mol/L. Under the optimal culture conditions, the diameter of haemolysis zone increased 29.3 % than before. HPLC analysis showed the precise production of lipopeptide was 30.2% higher than preliminary culture. Furthermore, at three batches cultivation, the experiment values under the optimal conditions agreed with the predictive values. It showed that Response Surface Methodology was proper and a good choice for optimization.

To control the quality of Humulus scandens, the quality standard was established in this study. According to the method recorded in the Appendix of Chinese Pharmacopoeia (2010 Edition) , the water and ash inspections were carried out. The component luteoloside and cosmosiin in Humulus scandens were identified and assayed by TLC and HPLC. The results showed a strong characteristics microscopic of Humulus scandens, and trichoromethane-methanol-formic acid (10: 3: 0. 3) as the mobile phase of TLC, the spots at 365 nm with a UV lamp was clear. The 16 batches of samples were analyzed by HPLC with a gradient elution of acetonitrile and phosphate solution (0.2%) at a flow rate of 1.0 mL · min(-1) and detected at 350 nm. The content of luteoloside was 0.015%- 0.651% (average 0.148%); the content of cosmosiin was 0.003%-0.118% (average 0.036%). The linear calibration curve of luteoloside and cosmosiin was acquired in the ranges of 0.011-0.364 g · L(-1) (r = 1.000 0) and 0.003-0.096 g · L(-1) (r = 1.000 0), respectively. The average recovery was 100.5% and 98.5%, respectively. The methods are convenient and reliable, which can be ap- plied for quality assessment of Humulus scandens.
China ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; analysis ; standards ; Humulus ; anatomy & histology ; chemistry ; Quality Control

Through searching domestic and foreign medical journals in CNKI, Wanfang database, VIP database and Pubmed database from January of 2003 to November of 2013, 39 articles regarding musical electroacupuncture (MEA) were analyzed. The result showed that MEA was clinically used to treat neurological and psychotic disorders; because it was combined with musical therapy and overcame the acupuncture tolerability, and MEA was superior to traditional electroacupuncture. However, problems such as low research efficiency and the mechanism of MEA superiority and the musical specificity not being revealed by research design still exist. In future, large-sample multi-center RCT researches should be performed to clarify MEA clinical efficacy. With modern science and technology and optimized study design, guided by five-element theory of TCM, researches on different musical elements and characteristics of musical pulse current as well as MEA's correlation with meridians and organs should be studied, so as to make a further exploration on MEA mechanisms and broaden the range of its clinical application.
Acupuncture Points ; Bibliometrics ; Combined Modality Therapy ; Electroacupuncture ; Humans ; Music Therapy

Objective: To investigate the trend in incidence of hepatitis C in Jingzhou City, Hubei Province from 2008 to 2022, and to examine the age-period-cohort effect, so as to provide the basis for the formulation of hepatitis C prevention strategies. Methods: Demographic data and incidence data of hepatitis C in Jingzhou City from 2008 to 2022 were collected through the Chinese Disease Prevention and Control Information System, and the trend in incidence of hepatitis C was analyzed using average annual percent change (AAPC) and annual percent change (APC). The effects of age, period and cohort on the incidence of hepatitis C were examined with an age-period-cohort model. Results: The average annual incidence of hepatitis C in Jingzhou City from 2008 and 2022 was 20.26/105, with a male incidence of 20.04/105 and a female incidence of 20.47/105. The incidence of hepatitis C initially rose and then fell (AAPC=5.375%, P<0.05), with a rising trend from 2008 to 2018 (APC=13.370%, P<0.05) and a decreasing trend from 2018 to 2022 (APC=-12.231%, P<0.05). The incidence of hepatitis C appeared a tendency towards a rise with age, and the 80-84 age group had the highest risk (RR=11.420, 95%CI: 7.631-17.090) in relative to the 45-49 age group. The incidence of hepatitis C appeared a tendency towards a rise followed by a decline with time, and an increased risk of hepatitis C was seen from 2013 to 2017 (RR=1.393, 95%CI: 1.272-1.525) and a decreased risk was seen from 2018 to 2022 (RR=1.237, 95%CI: 1.072-1.428) in relative to the period from 2008 to 2012. The incidence of hepatitis C appeared a tendency towards a rise followed by a decline with the cohort, and a higher risk was found in the 1965-1984 cohort (all RR>1.300) in relative to the 1960-1964 cohort. The incidence of hepatitis C, the age and period effects in men and women, and the cohort effects in men were consistent with the whole population. In addition to the 1965-1984 cohort, a higher risk was found in the 2000-2014 cohort in women (all RR>1.250). Conclusions From 2008 to 2022, the incidence of hepatitis C in Jingzhou City experienced a notable rise and subsequent decline. The incidence of hepatitis C increased with age, with higher risks seen among middle-aged and elderly people.

OBJECTIVETo observe the clinical efficacy of acupuncture at Hegu (LI 4) on central facial nerve paralysis after ischemic stroke, and explore dose-effect relationship among different stimulation intensities of acupuncture at Hegu (LI 4) as well as its optimal treatment plan.

METHODSAccording to different acupuncture stimulation intensities which were based on treatment time and needle insertion direction, fifty patients were randomly divided into a Hegu 1 group, a Hegu 2 group, a Hegu 3 group, a Hegu 4 group and a control group, ten cases in each one. Different stimulation intensities of acupuncture at Hegu (LI 4) combined with facial paralysis acupoints, including Yingxiang (LI 20), Dicang (ST 4), Jiache (ST 6) and Quanliao (SI 18), were applied in Hegu 1 to 4 groups; meanwhile acupuncture at stroke acupoints, including Neiguan (PC 6), Shuigou (GV 26) and Sanyinjiao (SP 6), and medication treatment were adopted. Except acupuncture at Hegu (LI 4), the treatment of the control group was identical as Hegu groups. The treatment duration lasted for 14 days. The House-Brackmann facial never grading systems (H-B), Toronto facial grading system (TFGS), degrees of facial never paralysis (DFNP), facial disability index (FDI) and clinical efficacy were compared among groups.

RESULTS(1) Compared before the treatment, H-B, TFGS, DFNP and physical function score in FDI were all improved significantly in the Hegu 1 to 4 groups (all P < 0.05), but social function score in FDI was not obviously improved (all P > 0.05); all the scores in the control group were not evidently changed (all P > 0.05). (2) Compared with the control group, differences of H-B before and after treatment in the Hegu 1 to 4 groups, differences of TFGS in the Hegu 2 group and differences of DFNP in the Hegu 1 and Hegu 2 group were significantly improved (all P < 0.05). The differences of any scale among Hegu 1 to 4 groups were not significant (all P > 0.05), in which the most evident change was found in Hegu 2 group. (3) The total effective rate was 90.0% (9/10), 100.0% (10/10), 90.0% (9/10) and 80.0% (8/10) in Hegu 1 to 4 groups, which were significantly higher than 60.0% (6/10) in the control group (all P < 0.05).

CONCLUSIONAcupuncture at Hegu (LI 4) has affirmative clinical efficacy on central facial nerve paralysis after ischemic stroke, in which oblique insertion along the opposite direction of meridian for 5 s of twirling manipulation has the best clinical effect.

Acupuncture Points ; Acupuncture Therapy ; Adult ; Aged ; Facial Paralysis ; etiology ; therapy ; Female ; Humans ; Male ; Middle Aged ; Stroke ; complications

OBJECTIVETo clone tpn17 and tpn47 genes of Treponema pallidum and then construct their prokaryotic expression systems,to establish ELISAs based on rTpN17 and rTpN47 as antigens and to evaluate the sensitivity and specificity of the ELISAs for detection of serological diagnosis of syphilis.

METHODSThe whole length of tpn17 and tpn47 genes was amplified by PCR and then their prokaryotic expression systems were constructed. SDS-PAGE was used to measure the expression of the target recombinant proteins rTpN17 and rTpN47. Ni-NTA affinity chromatography was applied to extract rTpN17 and rTpN47, while Western blot was performed to determine the specific immunoreactivity of rTpN17 and rTpN47. By using rTpN17 and rTpN47 as the coated antigen, respectively, ELISAs (rTpN17-ELISA and rTpN47-ELISA) were established to detect serum samples from 200 healthy individuals, 25 RA patients, 17 SLE patients and 211 syphilis patients. The detection effects of the ELISAs were compared to those of TRUST and TPHA.

RESULTThe sequence similarity of the cloned tpn17 and tpn47 genes was 100 % compared with the corresponding sequences in GenBank. The expression outputs of rTpN17 and rTpN47 were approximately 37.2 % and 26.8 % of the total bacterial proteins, respectively. Both the extracted rTpN17 and rTpN47 could take place remarkable conjugation reactions to the sera with positive antibody against Treponema pallidum.The positive detection rate of TPHA (99.1%) was the highest (P<0.001). The positive detection rates of rTpN17-ELISA (85.3 %) and rTpN47-ELISA (84.3 %) were similar (P>0.05). The positive detection rates of TRUST (72.5 %) was lower than that of rTpN17-ELISA (P=0.001) but similar to that of rTpN47-ELISA (P=0.014). The detection results of all the serum samples from healthy individuals, RA patients and SLE patients were negative, whereas 7.1 % (3/42) of the samples from RA or SLE patients were positive.

CONCLUSIONrTpN17 and rTpN47 are still maintaining their original immunoreactivity. The ELISAs using rTpN17 or rTpN47 as the antigen are rapid, simple and convenient, higher sensitivity and specificity methods for serological screening and detection of syphilis.

Antibodies, Bacterial ; Antigens, Bacterial ; Blotting, Western ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Humans ; Male ; Syphilis ; diagnosis ; Syphilis Serodiagnosis ; Treponema pallidum ; chemistry ; immunology ; isolation & purification

OBJECTIVETo search for the best therapy for slow transmission constipation (STC) or mixed constipation (MC) by intensive acupoints thread embedding (IATE).

METHODSA total of 310 patients with constipation were assigned to 5 groups using a stratification control method. The 62 patients of STC in the observation group I were treated by IATE at Dachangshu (BL 25), Tianshu (ST 25), Zhongji (CV 3) and Zusanli (ST 36) acupoints; the 62 of MC in the observation group II by IATE at the same acupoints and local anal operation; both the 62 of STC in the control group I and the 62 of MC in the control group 1I were treated by oral administration of Simo Decoction, while the 62 of MC in the control group III were treated by local anal operation alone. Conditions of defecation were recorded every day in the 6-month follow-up period. Gastrointestinal transit test (GITT) was carried out before and after treatment. One follow-up was conducted at the 1st, 3rd and 6th month.

RESULTSIn the 124 patients of the two observation groups, 52 (41.9%) were cured after treatment, they could normally discharge stool, 5 (4.0%) showed markedly effective with their constipation alleviated, but having occasional bowel slackened sensation. In the control group I and II, none was cured or effectively treated, while in the control group III, the efficacy in 8 patients were judged as marked effective, but none was cured.

CONCLUSIONIATE is a convenient and effective composite therapy. It shows favorable clinical efficacy for STC, and also for MC in case of combined with local anal operation.

Acupuncture Points ; Aged ; Anal Canal ; surgery ; Catgut ; Constipation ; therapy ; Female ; Humans ; Male ; Middle Aged ; Prosthesis Implantation ; methods

To investigate the effect of flavonoids from Sophora flavescens in aging mice induced by D-galactose and its mechanism. Totally 60 mice were randomly divided into six groups: the control group, the model group, the piracetam group (positive control group) and flavonoids from S. flavescens low, medium and high doses groups. Except for the control group, all of the rest groups were subcutaneously injected with D-galactose (160 mg x kg(-1)) for successively 30 days to establish the sub-acute senescent model. Meanwhile, flavonoids from S. flavescens low, medium and high doses groups were respectively administered with 150, 300 and 600 mg xkg-('1)of flavonoids from S. flavescens for 30 days. The learning and memory abilities of mice were determined by avoiding darkness ex-eriment and jumping stair experiment. The contents of malondialdehyde (MDA) tumor necrosis factor-aα NF-aα the activities of superoxide dismutase (SOD) monoamine oxidase-B (MAO-B) Na'(+)K'(+)-ATPase and Ca2(+ )-ATPase in the brain of mice were deter-ined respectively after the behavioral experiments. The activity of lactic dehydrogenase ( DH) in blood serum was also determined and analyzed by microscope after HE staining to observe the changes in hippocampal organizational structure. Compared with the model group, flavonoids from S. favescens medium and high doses groups showed significantly increases in the latency of avoiding darkness and jumping stair experiments; flavonoids from S. fllvescens low, medium and high doses groups and the piracetam group showed de-reases in the numbers of errors in avoiding darkness experiment; the flavonoids from S. flavescens high dose group and the piracetam group showed reduction- n the number of errors in jumping stair experiment (P <0 . 5 or P <0 . 1). Flavonoids from S. flavescens me-ium and high doses groups and the piracetam group showed improvements in the activities of SOD, Na'(+)K'(+)ATPase in the brain of mice and declines in the contents of MDA and TNF-aα the activity of MAO-B in the brain of mice, the activity of LDH in blood serum (P <0 . 5 or P <0 . 1). Flavonoids from S. flavescens low, medium and high doses groups and the piracetam group also showed im-rovement in the activity of Ca2(+ )ATPase, with statistical difference from the model group (P <0 . 5 or P <0 . 1). The pathological result showed decreases in the number of cells of hippocampal dentate gyrus area, sparse cell arrangement, incomplete cellular mor-hology, scarce cytoplasm, blurred boundary between nucleus and cytoplasm, nuclei anachromasis, irregular pyknosis and unconspicu-us nucleoli in the model group. Compared with the model group, flavonoids from S. flavescens low, medium and high doses groups and the piracetam group showed improvements in hippocampal organization tissues. Flavonoids from S. favescens can improve the learning and memory ability of senescent mice induced by D-galactose. Its mechanism may be correlated with the enhancement of anti-oxidative actions by lowering TNF-aαcontent, which results in the stability of cell membrane and the reduction in MAO-B activity.
Aging ; drug effects ; metabolism ; psychology ; Animals ; Brain ; drug effects ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Flavonoids ; administration & dosage ; Galactose ; adverse effects ; Hippocampus ; drug effects ; metabolism ; Humans ; Learning ; drug effects ; Male ; Malondialdehyde ; metabolism ; Mice ; Sophora ; chemistry ; Superoxide Dismutase ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Background The sex hormones plays an important role in the incidence of dry eye,especially for the regulation of function.However,the effects of sex hormones on lacrimal gland epithelial cells are below understand.Objective This study was to investgate the effects of estradiol and testosterone on the apoptosis of lacrimal gland cells induced by H2O2.Methods The lacrimal gland tissue was obtained from 2- or 3-month-old clean male New Zealand rabbits and the lacrimal gland epithelial cells were cultured in vitro using esplant culture method.The cells were identified by pan cytokeratin antibodies with immunocytochemistry.lacrimal gland epithelial cells were incubated in the 96 well plate at the density of 5 × l04 cells/ml for 44 hours.Estradiol or testosterone with the concentrations of 1 × 10-5,1 × 10-6,1 × 10-7,1 × 10-8 mol/L were added into the medium for 24 hours respectively and 1× 10-4 mol/L H2O2 treated the cells for 1 hour to induce the apoptosis in experimental groups.The cells treated by only 1 × 10-4 mol/L H2O2 were used as apoptotic control group,and the cells cultured by regular method were used as blank control group.The cell viability in different groups was detected using MTT at 570 nm ( A570 ),and the apoptotic rates of the cells were assayed using Annexin V/PI double staining.This use and maintain of experimental animals followed the Regulation for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The cultured cells showed the irregular polygon in shape,and about 80％ cells was positive response for cytokeratin.MTT assay showed that the lower A570 values were detected in the H2O2-induced group,various concentrations of estradiol or testosterone groups compared with blank control group (P＜0.01 ).The A570 values in 1 × 10-5,1 × 10-6,1 × 10-7 mol/L estradiol groups or 1 × 10-6 mol/L testosterone group were significantly higher than ones of H2 O2-induced group (P＜0.01 ).Compared with corresponding concentrations of testosterone groups,the A570values in various concentrations of estradiol groups were elevated( P＜0.01 ).The apoptosis rates at the early and later phase were significantly declined in both estradiol group and testosterone group in comparison with H2 O2-induced group (P ＜ 0.01,P＜ 0.05 ),and those in estradiol group were lower than the testosterone group( P＜0.01,P＜0.05 ).Conclusions Estradiol and testosterone suppress the apoptosis of lacrimal gland cells induced by H2O2,and the stronger effect is found in estrogen.The inhibition of estrogen on lacrimal gland cell apoptosis show a dose-dependent manner to some extent.