To set up a new method of detecting bacterial number in situ,NADH fluorescence method based on the fluorescent characteristic of NADH was used.When the concentration of NADH ranged from 10 nmol/L to 0.2 mmol/L,its concentration had a good line relationship to the fluorescence intensity(R2= 0.9905).Separating bacterial cells by centrifugation and extracting NADH with hot Tris-HCl buffer,the re-sult of bacterial count detected with NADH standard plot was 1?104 CFU/mL in an hour.In summary,NADH fluorescence method is rapid,sensitive,simple and reliable to detect total bacterial number.There-fore,the method can be widely applied in the field of food sanitation and safety,environment detection and so on.

Aged ; Cause of Death ; Humans ; Laryngeal Neoplasms ; mortality ; Male ; Peptic Ulcer Hemorrhage ; etiology ; mortality ; Postoperative Period

Adult ; Female ; Hemofiltration ; Hemolysis ; drug effects ; Humans ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Middle Aged ; Plasma Exchange ; Treatment Outcome ; Wasp Venoms ; poisoning ; Young Adult

AIM: To affirm marker peaks for the fingerprint chromatography of Shengmai Injection. METHODS: LC-MS/MS method was used, with a Waters symmetryshield TM RP_ 18 column(4.6 mm?250 mm; 5.0 ?m), acetonitrile-water as a mobile phase, The detection wavelength was at 203 nm. Ion trap mass spectrum. RESULTS: Affirming marker peaks for fingerprint chromatography of Shengmai Injection and 10 marker peaks were affirmed. CONCLUSION: The method can affirm marker peaks for the fingerprint chromatography of Shengmai Injection. It is simple, accurate, and has practicality.

AlM: To evaluate the therapeutic effects and safety of femtosecond laser assisted penetrating keratoplasty ( FS-PKP) .
METHODS: ln the retrospective case series study, 46 cases ( 48 eyes ) with FS- PKP holds were analyzed. Preoperative best corrected visual acuity ( BCVA) was 2. 35 ± 1.01 (with a standard logarithmic eye chart). Opacities were all from epithelium to endothelium, including keratoconus, keratolukoma and corneal scarring etc. The full-thickness corneal donor-recipient grafts in different diameter and incision kerfangle were made by femtosecond laser in keratoplasty. The patients were treated according to the conventional penetrating keratoplasty after surgery. Evaluation indexes included preoperative and postoperative 1, 6, 10mo uncorrected visual acuity (UCVA), BCVA, corneal astigmatism (CA) and surgical complications. Forty-three cases (44 eyes) with traditional PKP were as control group. The data of two groups were analyzed by mean comparison of two groups and Chi-square test in SPSS 18. 0.
RESULTS: All the operations were completed successfully, postoperative visual acuity of all patients had different degrees of increase. Comparison with the postoperative UCVA of traditional PKP surgery, UCVA was 3. 48 ± 0. 43 and 2. 97 ± 1. 14 at 1mo after operation, respectively, with statistically significant difference ( P=0. 009). The UCVA was 3. 86±0. 25 and 3. 74±0. 73 at 3mo after operation, respectively; the UCVA was 4. 04 ± 0. 23 and 3. 98 ± 0. 29 at 6mo after operation, respectively, with no statistically significant difference (P=0. 146, P=0. 56). ln FS-PKP group, BCVA was better than traditional PKP surgery group, with statistically significant difference ( P=0. 002 ). While there had no statistically significant difference between two groups of BCVA at 6, 10mo after operation (P6mo =0. 132, P10mo =0. 47). The incidence of complication of FS-PKP surgery, such as postoperative astigmatism, postoperative intraocular pressure and postoperative incision dehiscence were significantly lower than the one of traditional PKP surgery(P<0. 05).
CONCLUSlON:Compared with traditional penetrating keratoplasty, FS-PKP provides significantly faster visual recovery in the early postoperative period, and induces significantly less corneal astigmatism and lower incidence of complication. lt reflected the clinical superiority of this new technology .

To study the in situ intestinal absorption of five oligosaccharides contained in Morinda officinalis How. (sucrose, kestose, nystose, 1F-Fructofuranosyinystose and Bajijiasu). The absorption of the five oligosaccharides in small intestine (duodenum, jejunum and ileum) and colon of rats and their contents were investigated by using in situ single-pass perfusion model and HPLC-ELSD. The effects of drug concentration, pH in perfusate and P-glycoprotein inhibitor on the intestinal absorption were investigated to define the intestinal absorption mechanism of the five oligosaccharides in rats. According to the results, all of the five oligosaccharides were absorbed in the whole intestine, and their absorption rates were affected by the pH of the perfusion solution, drug concentration and intestinal segments. Verapamil Hydrochloride could significantly increase the absorptive amount of sucrose and Bajijiasu, suggesting sucrose and Bajijiasu are P-gp's substrate. The five oligosaccharides are absorbed mainly through passive diffusion in the intestinal segments, without saturated absorption. They are absorbed well in all intestines and mainly in duodenum and jejunum.
Animals ; Drugs, Chinese Herbal ; chemistry ; pharmacokinetics ; Intestinal Absorption ; Intestine, Small ; metabolism ; Male ; Morinda ; chemistry ; Oligosaccharides ; chemistry ; pharmacokinetics ; Perfusion ; Rats ; Rats, Sprague-Dawley

With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals ; Antibodies, Monoclonal ; chemistry ; genetics ; immunology ; Humans ; Hybridomas ; metabolism ; Immunologic Techniques ; methods ; trends

OBJECTIVETo investigate the relationship between the renal tubular function and the severity of tubulo interstitial lesion and the effects of Astragalus Injection (AI) on renal tubular function in patients with IgA nephropathy (IgAN).

METHODSSixty-seven patients with IgAN were randomly divided into the control group and the astragalus group, both received dipyridamole and benazepril orally, while the astragalus group treated with AI by intravenous dripping additionally. The indices for renal tubular function, including protein in blood and urine, urinary retinol binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (UNAG), were detected. Area of glomerular Bowman capsule, renal tubules, and capillary were measured with color magic image analysis system type CMIAS2000.

RESULTSUrinary RBP and UNAG were correlated with tubulointerstitial lesion. Urine protein concentration decreased, blood albumin increased remarkably and renal tubular function improved after treatment in the astragalus group, with the improvement significantly different to those in the control group respectively.

CONCLUSIONThe severity of tubulointerstitial lesion was positively related to urinary RBP concentration, and astragalus injection has obvious effect on IgAN.

Adolescent ; Adult ; Astragalus membranaceus ; Benzazepines ; therapeutic use ; Dipyridamole ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Glomerulonephritis, IGA ; drug therapy ; physiopathology ; Humans ; Injections, Intravenous ; Kidney Tubules ; physiopathology ; Male ; Middle Aged ; Phytotherapy

Objective To explore the reliability of relevant electrocardiogram(ECG) indexes in evaluating isoprenaline(ISO)-induced rat acute ischemic myocardial injury and provide reference for future scientific applications of these models. Methods Seventy male Wistar rats were randomly equally assigned to ten groups according to their body weight: 5,10,20,40,80,160,320,640, 1280 and 2560 μg/kg dose groups. All rats were tail intravenously given corresponding doses of saline diluted isoprenaline according to their body weight. Standard limb Ⅰ , Ⅱ, Ⅲ-lead ECG of all rats were recorded before, immediately after and 1,24,and 72 hour after injection, respectively.Changes of heart rate, T-wave amplitude of Ⅱ -lead and Q-T interval were measured. Results Significant differences were found in heart rates, T-wave amplitudes and Q-T intervals at different time points(F = 15.03,11.28,13.64, all P ＜ 0.01 ), while differences among the ten ISO-dose groups were statistically insignificant (F= 1.45, 1.17,1.09, all P ＞ 0.05). No interaction between observation time and ISO dose was observed on heart rates, T-wave amplitudes and Q-T intervals(F= 0.79,0.82,0.59, all P ＞ 0.05). Immediately after injection of ISO, the heart rates were significantly increased compared with that of pre-injection in all groups(all P ＜ 0.05), of which 320 and 640μg/kg dose groups increased most significantly [(550 ± 47), (521 ± 43)times/min]. T-waves decreased significantly compared with that of pre-injection (all P ＜ 0.01 ), and 20 μg/kg dose and above groups decreased particularly evident, and partly inverted. Q-T intervals of rats in each group were significantly shorter than that of pre-injection(all P ＜ 0.01 ), and 320, 640, 1280 μg/kg groups shortened more pronounced[(0.070 ± 0.006),(0.072 ± 0.005), (0.068 ± 0.005)ms]. One hour after injection, the heart rate of rats in each group decreased,except 320 and 640 μg/kg dose groups[(518 ± 43), (487 ± 36)times/min], which were still higher than that of pre-treatment[(450 ± 40), (448 ± 51 )times/min, all P ＜ 0.05], the rest groups no longer had significant differences (all P ＞ 0.05). ECG T-wave in each group was significantly recovered compared with that of instantly medication (all P＜0.05), and 40 μg/kg dose and above groups recovered more than a big margin, but there were still differences compared with that of pre-treatment (P ＜0.05), while T-waves of 40 μg/kg dose and below groups had returned to the level of pre-treatment. Q-T interval in each group had varying degrees of recovery, except 1280 and 2560 μg/kg dose groups[(0.080 ± 0.004), (0.076 ± 0.011 )ms]which were still less than that of pre-treatment[(0.086 ± 0.007),(0.085 ± 0.006)ms, all P ＜ 0.05], other groups had no significant difference compared with that of pre-treatment (all P ＞ 0.05). Twenty-four hours after injection of ISO, the heart rates of 1280 and 2560 μg/kg dose groups [(389 ± 31 ), (398 ± 23)times/min]decreased significantly compared with that of pre-treatment[(427 ± 43), (438 ±26)times/min, all P ＜ 0.05], while other groups had returned to the level of pre-treatment. Seventy-two hours after injection of ISO, the heart rates, T-wave amplitudes and Q-T intervals of all doses groups had returned to the level of pre-treatment (all P ＞ 0.05). Conclusions There has no significant ST segment in the electrocardiogram of rat.Isoprenaline has an exact effect on shortening Q-T interval. T-wave amplitude and Q-T interval can be used as reliable indexes of ECG for assessment of this animal model.

Objective To summarize the clinical characteristics of liver cirrhosis complicated with chylous ascites.Methods From July,2007 to November,2012,patients diagnosed as liver cirrhosis complicated with chylous ascites were retrospectively analyzed.Analyze the etiology,liver function,ascites,imaging examination,treatment options and progonsis of these patients.Differences in measurement data were compared with U test.Results A total of 34 cases were enrolled,male 27 cases and female seven cases,average age (51.7±12.5) years old.Hepatitis B (20/34,58.8％) was the most common etiology of liver cirrhosis.Child-Pugh grading was grade B (26 cases) and C (eight cases).The appearance of aseites of 17 cases was chylous and the serm-ascites albumin gradient (SAAG) was 18.8 (2.6 to 32.5) g/L.The SAAG of 27 cases (84.4％) was no less than 11.0 g/L.The ascites triglyceride (TG) level was 2.94 (0.26 to 16.75) mmol/L,the TG of 27 cases (84.4％) was no less than 1.25 mmol/L.The level of TG (2.66(0.26 to 16.75) mmol/L) of patients with SAAG over 11.0 g/L was significantly lower than that of SAAG lower than 11 g/L (7.07(2.26 to 15.67) mmol/L,U=24.00,P=0.02).The lymphatic scintigraphy indicated that imaging agent leaked into peritoneal cavity in 29 cases (85.3 ％).Direct lymphangiography revealed lymphatic vessel structure abnormality in 15 cases (60 ％).Twelve patients received conservative treatment and four patients were effective,13 patients accepted peritoneal-venous shunting and 12 patients were effective,11 patients with microsurgical treatrnent and seven patients were effective.Conclusions Lymphatic struture is a possible cause of cirrhosis patients with chylous ascites.The characteristics of chylous ascites in cirrhosis patients are still the same as the characteristics of portal hypertension ascites.The SAAG remarkably increases,and the the level of SAAG probably depends on TG level.Lymphoscintigraphy has great value on the determination of the presence of ascites.And the direct lymphangiography could help to reveal lymphatic vessel structure abnormality.