Objective To study the killing effect of polymorphonuclear neutrophils(PMNs) on Trichomonas vaginalis. Methods The vaginal secretion from a patient with vaginitis was incubated in the liver infusion liquid medium to get T. vaginalis. One ml serum was collected from the patient and heated for 30 min at 56 ℃ to inactivate complement in serum, and was absorbed three times with the parasites at 0 ℃ to make the serum free of antibodies. PMNs were separated from the patient’s blood and purified with density gradient centrifugation and polymer accelerating sedimentation. NBT and safranin O were used to stain the sample. The interaction between PMNs and the parasites was observed under microscope. 300 trichomonads and 3?104 PMNs were incubated for 10, 20, 30, 40, 50, 60 minutes under the conditions of aerobic or anaerobic, with superoxide dismutase (SOD) and catalase (CAT) or without SOD and CAT, and with complement or without complement. They were then inoculated in solid medium for another five days under the anaerobic condition, and surviving organisms were enumerated. Results PMNs were observed to surround and kill a single trichomonad. In the petri-dish containing PMNs, the surviving rate of the parasites in anaerobic condition was 85%, only 3% in aerobic condition (P

Objective To understand awareness of basic concepts, diagnosis, treatment and patient education for chronic obstructive pulmonary disease (COPD) among practitioners in rural new cooperative medical scheme (NCMS) and evaluate effectiveness of training for them in Haiyang, Shandong.Methods In total, 116 practitioners, two or three randomly selected from each of 43 NCMS service stations under the Third People's Hospital of Haiyang, were surveyed with questionnaire and then systematic training was provided for them. Another survey was conducted among them a half and two years after the first one with the same questionnaire. Results At the first survey, only 9.5 % (11/116) of the practitioners surveyed knew about "Guidelines for Prevention and Control of Chronic Obstructive Pulmonary Disease",19. 8 % (23/116) knew clinical characteristics of COPD, 29. 3 % (34/116) knew that pulmonary function test is gold standard for COPD diagnosis, 62. 9 % (73/116) knew that smoking is major inductive factor for COPD and could persuade patients to quit smoking, only 2.6 % (3/116) could make registration for the patients and provide health education for them, 48. 3 % (56/116) knew that the patients should be immunized with influenza vaccine, and 7. 8 % (9/116) knew long-term oxygen therapy for the patients. At the second survey, 94. 8 % (110/116) of the practitioners surveyed knew about " Guidelines for Prevention and Control of Chronic Obstructive Pulmonary Disease", all of them knew characteristics of COPD, that pulmonary function test is gold standard for COPD diagnosis, smoking is major inductive factor for COPD,could make registration for COPD patients and provide health education for them, 99. 1% (115/116) used influenza vaccine for COPD patients, and 99. 1% (115/116) knew long-term oxygen therapy for the patients. Conclusions Systematic training for COPD knowledge among practitioners in rural NCMS seems to be significantly beneficial for their awareness about the illness and to improve their abilities of prevention and treatment for it in rural areas.

Objective To study the antibacterial mechanisms of berberine and try to understand the reasons why bacteria cells difficultly resisted to it. Methods Detecting the minimal inhibitory concentration (MIC) of bacterial cultures incubated under sub-MIC concentration of berberine, Huanglian, and Neomycin for more than 200 generations, in order to analyze the bacteria resistance. Detecting the binding kinetics of berberine to DNA, RNA, and proteins. Observing the changes in bacterial cell surface structure with scanning electron microscopy. Detecting the Ca2+ and K.+ released from berberine-treated bacterial cells with atomic absorption spectrum. Detection the absorption of methyl-3H-thymine (3H-dT), 3H-uridine (3H-U), and 3H-tyrosine (3H-Tyr) into berberine-treated bacterial cells. Results MICs of bacterial cultures, growing more than 200 generations in MH medium with 1/2 MIC of berberine (BA200) or Huanglian (HA200), did not increase compared to the control, while remarkably increased in MH medium with 1/2 MIC of Neomycin (NA200). In addition, from the culture NA200 it was easy to isolate resistant mutant strains which could grow in MH medium with more than four times MIC Neomycin, but from the culture BA200 and HA200 it was difficult to isolate berberine or Huanglian mutant strains could grow in MH medium with more than four times MIC berberine or Huanglian. The binding kinetics of berberine to DNA, RNA, and proteins illustrated that berberine could easily and tightly bind to DNA and RNA, and hardly dis-bind from DNA- and RNA-berberine complexes. Berberine could easily bind to protein too, but also easily dis-bind from berberine-protein complex. The bacterial cells treated with berberine sharply decreased the absorption of 3H-dT, 3H-U, and 3H-Tyr, as the radioactive precursors of DNA, RNA, and protein biosynthesis. Berberine could damage bacterial cell surface structure, especially for Gram-negative bacteria. Ca2+ and K+ released from berberine-treated cells increased significantly compared to the control. Conclusion All of above results indicate that bacterial cells could not easily become resistant mutants to berberine. The mechanisms for the bactericidal effect of berberine include: inhibiting DNA duplication, RNA transcription, and protein biosynthesis; influencing or inhibiting enzyme activities; destructing the bacterial cell surface structure and resulting in Ca2+ and K+ released from cells. All of the berberine bactericidal mechanisms are the most essential physiological functions for a live cell, if influenced any one such function, the mutation would be lethal mutation, so that it is difficult to get berberine resistant cells. The results in this paper also prefigure that berberine and its related Chinese medicines would provide a feasible way to control antibiotic resistance problem.

Objective To investigate the effects of triptolide on lipopolysaccharide (LPS)-induced acute lung injury in rats.Methods Sixty-five male SD rats weighing 200-250 g were randomly divided into 5 groups: control group (group C,n =5),LPS group (group L,n =15),different doses of triptolide groups (groups TP1-3,n =15).In group C normal saline was injected iv and 1％ DMSO injected intraperitoneally(ip).In group L LPS 5 mg/kg was injected iv and 1％ DMSO injected ip.In groups TP1-3 LPS 5 mg/kg was injected iv and triptolide 25,50 and 100 μg/kg was injected ip respectively.Blood samples were collected at 1 h before administration and 1,3,6 and 12 h after administration for blood gas analysis.The animals were sacrificed at 12 h after administration.TNF-α concentrations in serum and bronchoalveolar lavage fluid (BALF) were determined by ELISA.The lungs were removed for microscopic examination,evaluation of diffuse alveolar damage (DAD) score and determination of W/D lung weight ratio and the expression of Toll-like receptor4 (TLR4) protein and mRNA.Results Compared with group C,PaO2 was significantly decreased at 3,6 and 12 h after administration,DAD score and W/D lung weight ratio were increased in groups L and TP1-3,TNF-α concentrations in serum and BLAF were increased,expression of TLR4 mRNA and protein was up-regulated in groups L,TP1 and TP2,whlie TNF-α concentrations in serum and BLAF were significantly decreased,expression of TLR4 mRNA and protein was down-regulated in group TP3 ( P ＜ 0.05).Compared with groups L and TP1,PaO2 was significantly increased at 6 and 12 h after administration,DAD score,W/D lung weight ratio and TNF-α concentrations in serum and BLAF were decreased,expression of TLR4 mRNA and protein was down-regulated in groups TP2 and TP3 ( P ＜ 0.05).There was no siginificant difference in blood gas parameters,DAD score and W/D lung weight ratio between group L and group TP1 and between group TP2 and TP3 ( P ＞ 0.05).Compared with group TP2,TNF-α concentrations in serum and BLAF were significantly decreased,expression of TLR4 mRNA and protein was down-regulated in group TP3 (P ＜ 0.05).The lung pathologic injury was reduced in groups TP1-3 as compared with group L.Conclusion Triptolide can attenuate acute lung injury induced by LPS in a dose-dependent manner in rats,and the inhibition of up-regulation of TLR4 expression and release of TNF-α may be involved in the mechanism.

AIM Objective By comparing E.coli's resistance to antibiotics and Chinese medical herbs before and after incubated in LB containing Neomycin or Chinese medical herbs, we try to understand the difference in their mechanism of resistance. METHODS We incubated E.coli cells under the culturing media with low concentration of Neomycin and Chinese medicine herbs respectively; lately, the culturing process were continued under the media with no Neomycin or Chinese medicine herbs. We tested the minimal inhibitory concentrations(MIC)of Neomycin(Kanamycin, Gentamicin, Tetracyclin etc) and Chinese medical herbs to culture cells. Result The experimental data indicated that increased resistance of population cells in the broth including Neomycin happened easily, as well as multi resistance simultaneously. As for the Chinese medical herbs, the case is different. After growing in the LB containing one of Huang Lian (rhizoma coptidis), Berberine or San-Huang-Tang of low concentrations, population cells did not increase their resistance either to antibiotics or to Chinese medical herbs. Removing the pressure of Neomycin, E.coli population cells resistance came back to the initial level. CONCLUSION The increased antibiotic resistance of population cells is unstable, and alternative using of different antibiotics maybe contribute to the alleviation of antibiotics resistance.

Aim To assay the mutagenicity of Enphorbia lunulata(EL) decoction and to modify the Ames test for evaluation the mutagenicity of herbal medicine samples.Method The mutagenicity of EL decoction was assayed by standard Ames test; the teratogenicity was done by mammalian bone marrow chromosomal aberration test. In modified Ames test system,the influence of histidine EL decoction was excluded by additional negative control, in which the test media was supplied with histidine (histidine amount equaled to the histidine in different concentration of EL decoction).Result The mutagenicity of EL decoction was positive in the standard Ames test. The teratogenicity of EL decoction was negative in mammalian bone marrow chromosomal aberration test. By the modified Ames tests,the mutagenicity of EL decoction was negative.Conclusion The standard Ames test is not suitable for evaluating the mutagenicity of EL decoction, but the modified Ames test is. The mutagenicity in vitro and the teratogenicity in vivo of EL decoction are all negative.

ObjectiveTo study the modes of case finding and diagnosis for inpatients of active pulmonary tuberculosis with primary treatment.MethodsData of 1000 inpatients with active pulmonary tuberculosis input into a computer were analyzed retrospectively.including clinical symptoms,signs and relevant laboratory examinations.to evaluate their diagnostic value.ResultsAmong 1000 active tuberculosis case8 hospitalized with symptoms and signs,95.9 percent suffered by cough,77.7 percent by expectoration and 50.8 percent (n=508) by fever,and 51.5 percent (n=777) with strong positive purified protein defivative (PPD) skin test,61.5 percent with positive serum anti-tuberculosis antibody,48.8 percent with positive acid-fast staining on sputum smear and 57.9 percent with positive sputum bacteriologle culture.And,49.4 percent of the patients were diagnosed by laboratory positive sputum,and 50.6 percent of those with negative sputum were diagnosed by comprehensively clinical considerations,ineluding 51.6 percent positive PPD skin teat, or positive serum anti-tubereulosis antibody,but 48.4 percent of them were all negative in varied laboratory examinations.ConclusionsHospital visit due to symptom is the main method for tuberculosis finding in our country.All those with Cough for two or more weeks should be screened by routine examinations for excluding tuberculosis.Case finding rate Was low by sputum examinations.so comprehensive diagnosis is still important for those tuberculosis patients with negative sputum.

Objective To evaluate the effect of lipopolysaccharide (LPS) on the expression of ATP-binding cassette transporter A1 (ABCA1) in alveolar macrophage cells of rats.Methods The alveolar macrophage cells of rats NR8383 were seeded in 6-well plates at a density of 1 × 106 cells/ml (2 ml/well) and randomly divided into 6 groups:control group (group C,n =24),0.2 μg/L LPS group (group L0.2,n =12),2.0 μg/L LPS group (group L2.0,n =12),20.0 μg/L LPS group (group L20.0,n =60),200.0 pg/L LPS group (group L200.0,n =12),and 20.0 μg/L LPS + ABCA1 siRNA group (group L20.0 + siRNA,n =12).The cells were routinely cultured in group C.LPS with the final concentrations of 0.2,2.0,20.0 and 200.0μg/L was added to the culture medium in L0.2,L2.0,L20.0 and I200.0 groups,respectively.In group L20.0 + siRNA,siRNA 50 nmol/L was added to the culture medium and 12 h later LPS 20.0 μg/L was added.In group C,6 wells were chosen for determination of ABCA1 mRNA and protein expression.At 24 h of incubation with LPS 0.2,2.0 and 200.0 μg/L,or at 2,6,12 and 24 h of incubation with LPS 20.0 μg/L,6 wells were chosen and the cell suspension was obtained for measurement of ABCA1 mRNA expression (by real-time fluorescent quantitative PCR),and ABCA1 expression (by flow cytometry).At 12 h of incubation with 20.0 μg/L LPS or with 20.0 μg/L LPS + 50 nmol/L siRNA,6 wells were chosen and the cell suspension was obtained for measurement of TLR4 mRNA expression (by real-time fluorescent quantitative PCR) and TLR4 expression (by flow cytometry).Results Compared with group C,the expression of ABCA1 mRNA and protein was significantly down-regulated in L0.2,L2.0,L20.0 and L200.0 groups,and the expression of ABCA1 mRNA and protein was up-regulated in L20.0 and L20.0 + siRNA groups.In L20.0 group,the expression of ABCA1 mRNA and protein was gradually down-regulated with the prolonging time of incubation with LPS.Compared with group L20.0,the expression of TLR4 mRNA and protein was significantly up-regulated in group L20.0 + siRNA.Conclusion LPS can down-regulate the expression of ABCA1 in alveolar macrophage cells of rats,however,ABCA1 can inhibit the synthesis of TLR4.

Background To study the pathogenesis and management of diabetic retinopathy (DR) has an important clinical significance.With the development of biomedical photonics in recent years,photobiomodulation therapy has been paid more and more attention.However,the sudy on biological regulation of light to DR is rarely reported.Objective This study was to explore the photobiomodulating effects on the apoptosis of retinal neuronal cells induced by high glucose environment and tried to offer a basis for the management of DR.Methods The retinal neurons were isolated from Wistar rats using immunomagnetic beads and primarily cultured in Neurobasal,and the cells were identified by Nissl staining.The cells were divided into normal control group,high-glucose group and high-glucose+LED group.The glucose at the concentration of 25 mmol/L was added into medium for 48 hours in the high-glucose group,and the cells induced by high-glucose were irradiated in incubator by LED for consecutive 300 seconds per time in a 12-hour interval with the wavelength of 620 nm,maximal power of 1 W,central light radiation exposure of 6.67 mW/cm2 and spot diameter of 2.0 cm.The apoptosis rate of the cells was assayed by flow cytometry;the intracellular Ca2+ content was determined by laser scanning confocal microscope;the relative expression level of phosphorylated serine-threonine kinase (p-AKT) protein in the cells was detected by Western blot.Results The cells grew well 2-3 days after cultured with the polygon and oval shape,and nucleolus were visible.More neuronal processes were obtained in 5-7 days after culture.Nissl staining showed the blue violet color in cytoplasma of neurons.The proportion of neurons and glial cells was 91％.The apoptosis rates of the cells were (7.634±3.176)％,(33.642 ±9.315)％ and (23.914±6.375)％ in the normal control group,high-glucose group and high-glucose+LED group,respectively,and the apoptosis rates of high-glucose group and high-glucose + LED group were significantly higher than that in the normal control group,while the apoptosis rate in the high-glucose+LED group was lower than that in the high-glucose group (all at P＜0.01).The fluorescence of Ca2+ in the cytoplasma was strong in the highglucose group and weak in the normal control group.The fluorescence pixel values in the high-glucose group and highglucose+LED group were significantly higher than that in the normal control group,and that in the high-glucose+LED group was reduced in comparison with high-glucose group (all at P＜0.05).The expressing band of p-AKT protein was strong in the normal control group and weak in the high-glucose group.The relative expressing levels were 10.34± 3.18,2.16±0.46 and 7.15 ±1.72 in the normal control group,high-glucose group and high-glucose+LED group,and relative expression level of high-glucose+LED group was significatly lower than that in the high-glucose group (P＜ 0.05).Conclusions High-glucose environment inhibits PI3K/AKT pathway and calcium homeostasis of retinal neurons,which results in cell apoptosis.Low intensity of LED light irradiation activates the anti-apoptotic PI3K/AKT pathway and therefore reduces apoptosis induced by high glucose.