AIM:To study the feasibility of recombinant adeno-associated virus(rAAV)as a vector to transfer the green fluorescent protein(GFP)gene as a target gene into rabbit retina.METHODS:Intravitreal injection of rAAV-gfp was performed in either eye for each rabbit with the other eye taken as control.At the 3rd,7th,and 14th day after injection,the eyeballs were removed,and the retinas were flat-mounted on glass slides to inspect the retinal fluorescence,respectively.RESULTS:After intravitreal injection of rAAV-gfp,the presence of fluorescent spots in the cytoplasm of retinal cells indicated that GFP gene was efficiently transferred and expressed in the rabbit retina.CONCLUSION:Recombinant adeno-associated virus is a reliable and simple vector for transferring target gene,e.g.,GFP gene,to the retina.

Animals ; Cold Temperature ; adverse effects ; Heart ; physiopathology ; Inflammation ; physiopathology ; Myocardium ; metabolism ; Oxidative Stress ; Particulate Matter ; adverse effects ; Rats

A reliable method for simultaneous determinition of eleven representative components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin) in combination of chromatographic fingerpint analysis for Reduning injection was developed by ultra high-performance liquid chromatography (UPLC). The method was performed on an Agilent ZORBAX SB-C18 anlytical column (3. 0 mm x 100 mm, 1. 8 µm) with a guard column of Agilent UPLC Guard ZORBAX SB-C18 (3.0 mm x 5 mm) at the column temperature of 30 °C. The gradient mobile phase consisted of acetonitrile (A)-0. 1% phosphoric acid (B) with a flow rate of 0. 4 mL . min-1. The injection volumn was 2 µL. The detection wavelengths were set at 324 nm and 238 nm for quantit tive analysis and 225 nm for fingerpint chromatography. Neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, shanzhiside, geniposidic acid, genipin-1-β-D-gentiobioside, geniposide and secoxyloganin were baseline seperated with good linearity relationships (r >0. 999) between concentration and peak areas over the linear ranges. The average recoverys of the investigated compounds were 103.5%, 100. 2%, 103. 3%, 102. 8%, 101. 3%, 102. 8%, 97. 36%, 99. 62%, 98. 16%, 102. 8%, 99. 27%, respectively. Reduning injection of forty-five batches was analyzed by UPLC finge print chromatography. Thirty batches were selected to generate the reference fringerprint chromatography with fourteen common peaks. The similarity values between the reference fringerprint chromatography and the remaining fifteen batches were higher than 0. 99. The developed method was fast, accurate and sensitive. It could be used as a reference for the quality control of multiple components determination and fingerprint chromatography for Reduning injection in future.
Chlorogenic Acid ; chemistry ; isolation & purification ; standards ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; standards ; Glucosides ; chemistry ; isolation & purification ; standards ; Iridoids ; chemistry ; isolation & purification ; standards ; Quality Control ; Reference Standards ; Reproducibility of Results ; Sensitivity and Specificity ; Time Factors

OBJECTIVETo establish a rapid quantitative analysis method for the content of chlorogenic acid and solid content in the extraction liquid concentration process during the production of Reduning injection by using the near-infrared (NIR) spectroscopy, in order to reflect the concentration state in a real-time manner and really realize the quality control of concentrating process of the extraction and concentration process.

METHODThe samples during the Jinqing extraction liquid concentration process were collected. After the removal of abnormal samples, the spectra pretreatment and the wave band selection, the quantitative calibration model between NIR spectra and chlorogenic acid HPLC analytical value and solid content was established by using PLS algorithm, and unknown samples were predicted.

RESULTThe correlation coefficients between the chlorogenic acid content and the solid content were respectively 0.992 1 and 0.994 0, and the correlation coefficients of the verification model were respectively 0.994 4 and 0.998 4, with the root mean square error of calibration (RMSEC) of 0.814 6 and 2.656 1 and the root mean square error of prediction (RMSEP) of 0.704 6 and 1.876 7 respectively, and the relative standard errors of predictions (RSEP) were 6.01% and 2.93% respectively.

CONCLUSIONThe method is simple, rapid, nondestructive, accurate and reliable, thus could be adopted for the fast monitoring of the chlorogenic acid content and the solid content during the concentration process of Reduning injection extraction liquid.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control ; Spectroscopy, Near-Infrared ; methods

AIMTo study the effects of TNF receptor blocking peptide on adjuvant arthritis in rats.

METHODSThe model of rat adjuvant arthritis was induced by injection of complete Freund's adjuvant. The TNF receptor blocking peptide was injected locally in the ankle. The ankle swelling, the pathologic changes in the ankle joint and the expression of IL-1 beta mRNA and TNF-alpha mRNA by peritoneal macrophages (RT-PCR) were observed.

RESULTSThe model of rat adjuvant arthritis induced by injection of complete Freund's adjuvant was similar to human rheumatoid arthritis. The treatment with TNF receptor blocking peptide for 10 days resulted in complete inhibition of joint swelling, a decrease in infiltration of inflammatory cell into joint tissue, an obvious alleviation of inflammatory pathological damages and an apparent decline of TNF-alpha mRNA and IL-1 beta mRNA of peritoneal macrophages of rats.

CONCLUSIONThe TNF receptor blocking peptide can protect the joint from inflammatory damage induced by adjuvant arthritis by suppression of TNF-alpha and IL-1 production, thereby alleviating the pathological injury of joint and controlling effectively the clinic course of arthritis.

Animals ; Ankle Joint ; pathology ; Arthritis, Experimental ; immunology ; pathology ; Interleukin-1 ; biosynthesis ; genetics ; Macrophages, Peritoneal ; metabolism ; Male ; Peptides ; pharmacology ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Receptors, Tumor Necrosis Factor ; antagonists & inhibitors ; Tumor Necrosis Factor-alpha ; biosynthesis ; genetics

Objective To investigate the effect of atorvastatin on oxidative stress and intracellular lipid accumulation in HepG2 cells under inflammatory state and explore the underlying mechanism.Methods HepG2 cells were treated with 100 ng · mL-1 TNF-α,100 ng · mL-1 TNF-α ± 10 μmol · L-1 atorvastatin in the presence of LDL for 24 h.Oil red O staining was used to examine the intracellular lipid contents.The mRNA and protein expressions of lipogenic genes (FAS,ACC and SREBP1) were detected by real-time polymerase chain reaction and Western blot.ROS levels were measured with the fluorescent probe of DCFH-DA.Contents of H2O2 and MDA were determined using the colorimetric method.Results Compared with normal group(the gray value of SREBP1 was 1.01 ± 0.001),the gray value of SREBP1 in model group was 1.61 ± 0.34.The mRNA levels in normal group of SREBP1,FAS,ACC respectively were 1.01 ± 0.16,1.03 ± 0.32,0.95 ± 0.29,the values in model group respectively were 3.61 ± 0.39,1.99 ± 0.36,2.37 ± 0.52,the differences were statistically significantly (P ＜ 0.05).Compared with model group,the mRNA levels of SREBP1,FAS,ACC and the gray value of SREBP1 in experimental group respectively were 2.95 ± 0.92,3.99 ± 1.16,2.85 ± 0.91,2.94 ± 0.65,the differences were statistically significantly(P ＜0.05).At the same time,compared with normal group,the levels of ROS(fluorescenceintensity),H2O2,MDA respectively were 1.00 ±0.20,and (2.30 ±0.31) (0.78 ±0.22) nmol · mg-1,the levels in model group respectively were 1.77 ± 0.25 and (4.32 ± 0.77),(1.86 ± 0.23) nmol · mg-1,the differences were statistically significantly (P ＜ 0.05).Compared with model group,the levels of ROS,H2 O2,MDA in HepG2 cells in experimental group respectively were 3.2 ±0.53 and (5.31 ±0.75),(3.43 ± 1.15) nmol · mg-1,the differences were statistically significantly(P ＜ 0.05).Conclusion Atorvastatin induced intracellular lipid accumulation in HepG2 cells under inflammatory stress,which may be associated with the increased oxidative stress.

OBJECTIVETo explore the expressions of endothelial nitric oxide synthase (eNOS) and cytochrome P450 (aromatase) in the testis of sexually mature male SD rats and their significance.

METHODSEighteen male SD rats, 6 five-week, 6 seven-week and 6 ten-week old, were selected for this study. Paraffin sections of the left testis were made and the expressions of eNOS and P450 observed by the immunohistochemical ABC method.

RESULTSPositive expressions of eNOS and P450 were found to be + + +, + and + + in the Leydig cells of the five-week, seven-week and ten-week old rats, respectively, and they were also observed in a few spermatocytes, though with no regularity.

CONCLUSIONIn the Leydig cells of sexually mature male SD rats, eNOS and P450 are differently expressed in different stages of sexual maturation, and they are correlated as well.

Animals ; Aromatase ; metabolism ; Male ; Nitric Oxide Synthase Type III ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sexual Maturation ; Testis ; metabolism

OBJECTIVETo explore the lower urinary tract symptoms (LUTS), especially those in the urinary storage phase, following transurethral resection of the prostate (TURP), and to improve the postoperative management and patients' quality of life after TURP.

METHODSA total of 86 patients with benign prostate hyperplasia (BPH) underwent TURP, and were interviewed on urinary symptoms at 1, 3, 7, 15 and 30 days after removal of the catheter. The patients were divided into two groups according to whether they had preoperative detrusor instability and/or compliance of the bladder (Group A) or not (Group B), and observed for the changes in IPSS scores and urinary storage symptoms after removal of the catheter.

RESULTSComplete follow-ups were achieved in 71 cases, 28 with detrusor instability and/or compliance of the bladder and the other 43 without. Their IPSS scores on the 1st, 3rd, 7th, 15th and 30th day after removal of the catheter were 8.1 +/- 2.5, 7.2 +/- 3.1, 6.3 +/- 3.8, 5.3 +/- 4.2 and 2.4 +/- 3.4, respectively, with statistically significant differences between the 7th and the 1st as well as the 30th and the 15th day (P < 0.05), but not between the 1st and the3rd nor the 15th and the 7th day (P > 0.05). On the 1st day, the cardinal symptoms in the urinary storage phase were urinary frequency, urgency and incontinence; the scores on IPSS and urinary storage symptoms were 10.4 +/- 3.3 and 9.3 +/- 3.8 in Group A and 6.2 +/- 2.8 and 5.2 +/- 2.7 in Group B, with significant differences between the two groups (P < 0.05). After treatment with tolterodine and alpha-adrenoreceptor inhibitor, neither IPSS scores nor the scores on urinary storage symptoms showed any significant differences between Groups A and B on the 15th and 30th day (P > 0.05).

CONCLUSIONThe lower urinary tract symptoms following TURP, especially those in the urinary storage phase, are correlated with preoperative bladder function, and getting improved gradually after surgery.

Aged ; Aged, 80 and over ; Humans ; Male ; Middle Aged ; Postoperative Period ; Prostatic Hyperplasia ; physiopathology ; surgery ; Quality of Life ; Transurethral Resection of Prostate ; Treatment Outcome ; Urinary Incontinence ; etiology

Objective To understand healthcare-associated infection(HAI)in group burn/trauma patients,and explore its risk factors.Methods HAI occurred among 25 hospitalized patients with group burn/trauma during a bus deflagration event on May 12,2014 were monitored and analyzed.Results Of 25 burn/trauma patients,7 de-veloped 10 times of HAI,incidence and case incidence of HAI were 28.00% and 40.00% respectively;the main in-fection sites were wound and lower respiratory tract,accounting for 60.00% and 30.00% respectively.30 strains of pathogens were isolated from 7 patients with HAI,including 16 strains (53.34%)of gram-negative bacteria,13 strains(43.33%)of gram-positive bacteria,and 1 fungus (3.33% ).Risk factors for HAI were large burned area, high degree of inhalation injury,arteriovenous catheterization,urinary tract catheterization,tracheotomy,use of ventilator,and surgery.Conclusion Incidence of HAI is high in the group burn/trauma patients,corresponding in-tervention measures should be formulated according to the risk factors,so as to reduce the occurrence of HAI.

Objective To investigate the value of endoscopy combined with laparoscopy in the treatment of colorectal polyps and polyp canceration.Methods Different combinations of endoscopic and laparoscopic procedures were employed and the clinical efficacies were compared.Results From January 2004 to September 2006,46 cases with colorectal polyp were treated with endoscopy combined with laparoscopy.Among them,5 cases(10.87%)underwent laparoscopic-assisted endoscopic polypectomy,30(65.22%)endoscopic-assisted laparoscopic resection,6(13.04%)synchronously endoscopic and laparoscopic resection.Five cases were performed further operation after endoscopic polypectomy.According to the pathological findings,21(45.7%)were proved to be polyp canceration,among which 6 were advanced carcinoma,and 3 were found metastasis to the lymph nodes.Among the 41 cases of laparoscopic resection,there was no conversion to an open surgery.Anastomotic leakage was found in 2 cases and anastomotic bleeding in 1.In the 5 cases of laparoscopic-assisted endoscopic polypectomy,no complication was observed.During the period of follow-up(1 to 21 months),no recurrence was detected.Conclusion Endoscopy combined with laparoscopy extends the safety and indications of endoscopic polypectomy,and is minimally invasive to the patients.It is an ideal procedure in the treatment of colorectal polyps and poly carceration.