OBJECTIVE:To prepare ganciclovir hydrochloride chitosan eye drops and establish its quality control method.METHODS:The eye drops was prepared using chitosan as thickening agent and synergist with ganciclovir as principal agent.The content of ganciiclovir was determined by HPLC.The stability of the preparation was investigated as well.RESULTS:The prepared eye drops was colorless transparent glutinous limpid liquid with its identification items all up to the related standards stipulated in Chinese Pharmacopoeia(2005 edition).The linear range of ganciclovir was 5～80 mg?L-1(r=0.999 9) with average recovery of 99.02%(RSD=0.71%).The preparation was stable under light with all its indexes showed no obvious change in the accelerated test at 6 month.CONCLUSION:The preparation technology is simple and feasible and the quality of preparation is stable and controllable with a validity duration tentatively set at 2 years.

OBJECTIVE: To compare the pharmacokinetics and relative bioavailability between ganciclovir-chitosan eye drops and ganciclovir eye drops in rabbit eyes. METHODS: Japan big-eared rabbits were randomly assigned to the receive 50 ?L ganciclovir-chitosan eye drops (experiment group) or ganciclovir eye drops (control group) in both eyes. The rabbit tears,corneas and aqueous humor were collected at different time for the determination of drug concentration by RP-HPLC. The pharmacokinetics parameters were calculated as well. RESULTS: In the experimenal group,the half-life(t1/2) in tears,corneas and aqueous humor were 15.41,45.21,59.43 minutes,respectively versus 16.22,32.55,and 48.53 minutes in the control group. The Cmax were 2.03 ?g?mL-1 and 1.22 ?g?mL-1,respectively. In the experimental group,the AUC0→240 in tears,corneas and aqueous humor were 1.66,1.89,and 2.77 times those in control group. CONCLUSION: The bioavailability of ganciclovir-chitosan eye drops was significantly higher than that of ganciclovir eye drops.

Objective:To study the influence of Wnt5a gene silence on the proliferation,migration and invasion of lung squamous carcinoma cells.Methods:A recombinant plasmid pHI-siRNA~(Wnt5a-)was constructed and used to deliver small interference RNA (siRNA)targeting Wnt5a in SK-MES-1 cells;the transfected cells were screened to establish a stable transgenic cell line.MTT,cell cycle and Transwell assays were employed to evaluate the effect of Wnt5a gene silence on the proliferation,migration and invasion of lung squamous carcinoma cells.Results:Western blotting assay revealed that Wnt5a was lowly expressed in SK-MES-1~(Wnt5a-)(13.6%).The proliferation index(PI)of transgenic cell line was slightly lower than that of the control cell line([28.3?3.8]% vs[30.5?5.2]%). The migration and invasion capabilities of SK-MES-1~(Wnt5a-)cells were(47.3?9.2)% and(39.7?11.7)% of the control cells, respectively.Conclusion:Low Wnt5a expression can significantly inhibit the migration and invasion capabilities of SK-MES-1 cells, indicating that Wnt5a might be a potential target for gene therapy of lung squamous carcinoma.

Objective To establish an effective PCR assay for leptospirosis detection , and applicate the assay in tree shrew, mongolian gerbil and gray hamster .Methods Sequence of leptospira was obtained from the NCBI Genbank , and primers were designed based on the sequences .The positive amplified fragments were sequenced to verify the reliability of the method.The samples from tree shrew, mongolian gerbils and hamsters were tested using this PCR method .Results The PCR method for detection of leptospirosis was successfully established .The positive rate of Leptospira was 8.33% in 60 samples of conventional tree shrews , 100% in 104 samples of the conventional Mongolian gerbils , and 0% in 60 samples of clean gray hamsters.Conclusions The establishment of this PCR assay is useful in the detection of leptospirosis in tree shrew, mongolian gerbil and gray hamster .The results of our investigation of leptospira infection levels of the three new experimental animals may promote their application in biomedical research .

BACKGROUND: This study was undertaken to measure the concentration of adiponectin (APN) in serum and induced sputum in patients with chronic obstructive pulmonary disease (COPD during acute exacerbation (AECOPD) and at stable stage and to determine the role of APN as a marker of inflammation in the pathogenesis of COPD. METHODS: All the patients in this prospective study were enrolled from October 2008 to October 2009, including 30 male AECOPD patients from the emergency department, 30 male stable COPD patients from the department of respiratory diseases, and 30 healthy non-smoking male controls from the department of medical examination. The serum and induced sputum were collected from each patient. All of the patients had normal weight (BMI range 18.5-24.9 kg/m2). Patients with severe bronchial asthma, bronchiectasis or autoimmune disease were excluded. Cell count and classification was performed for the induced sputum. The concentrations of APN, IL-8, IL-6 and TNF-α were measured by ELISA. Pulmonary function was tested among the three groups. Comparisons between the groups were conducted by Student's t test, ANOVA analysis or nonparametric test. Correlation analysis was carried out by Pearson's product-moment correlation coefficient test or Spearman's rank-order correlation coefficient test. RESULTS: The concentrations of APN in the serum or induced sputum in AECOPD patients were significantly higher than those in stable COPD patients or healthy non-smoking controls (P<0.01). The concentration of APN in stable COPD patients was significantly higher than that in healthy non-smoking controls (P<0.01). For the AECOPD patients, APN was positively correlated with IL-8 and TNF-α in the serum and induced sputum (r=0.739, 0.734, 0.852, 0.857 respectively, P<0.05). For the stable COPD patients, APN was also positively correlated with IL-8 and TNF-α in the serum and induced sputum (r=0.751, 0.659, 0.707, 0.867 respectively, P<0.05). In addition, for the AECOPD patients, APN was positively correlated with the percentage of neutrophils in the induced sputum (r=0.439, P<0.05). CONCLUSIONS: APN is involved in the process of systematic and airway inflammation of COPD. This process is related to neutrophils in the airway, IL-8 and TNF-α. APN could be used as a new marker for inflammation of COPD.

To study the protective effect and the mechanism of asiatic acid (AA) from Potentilla chinensis on alcohol hepatic injury in rats. Male Wistar rats were randomly divided into six groups: the normal control group, the AA control group (8 mg · kg(-1) AA), the model group (5.0-9.0 g · kg(-1) alcohol) and high, medium and low-dose AA-treated groups (alcohol + 8, 4, 2 mg · kg(-1) AA). Each group was orally administered with the corresponding drugs once a day for 24 weeks. Approximately 1. 5 hours after the final administration, all rats were killed, and their blood samples and hepatic tissues were collected. The AST and ALT in rat serum and the contents of MPO, TNF-α, IL-1β, SOD, GSH-Px, GSH-Rd and MDA in hepatic tissues were detected. The expressions of NF-κB, TLR4, CD14, MyD88, TRIF and protein expression in hepatic tissues were measured by western blot. The pathological changes in liver tissues were observed by histological examination. The results showed that compared with the model group, the AA-treated groups showed significant decreases in serum ALT, AST and MDA and increases in the activities of SOD, GSH-Px, GSH-Rd and MPO. Moreover, AA markedly inhibited the expressions of TNF-α, IL-1β, TLR4, CD14, MyD88 and NF-κB. The histological examination showed alleviated hepatic issue ijury to varying degrees. In short, asiatic acid (AA) from P. chinensis could protect alcohol-induced hepatic injury in rats. Its mechanism may be related to the inhibition of NF-κB inactivation and the reduction of inflammatory response.
Animals ; Liver ; drug effects ; pathology ; Liver Diseases, Alcoholic ; prevention & control ; Male ; NF-kappa B ; physiology ; Pentacyclic Triterpenes ; pharmacology ; Potentilla ; chemistry ; Protective Agents ; pharmacology ; Rats ; Rats, Wistar ; Toll-Like Receptor 4 ; antagonists & inhibitors

BACKGROUND:Pulmonary infection is the main complication after kidney transplantation, and its onset and morbidity may be related to conventional oral drugs after kidney transplantation.
OBJECTIVE:To analyze the effect of methylprednisolone instead of prednisone on severe pulmonary infection after kidney transplantation.
METHODS:Clinical data of 58 patients with severe pulmonary infection after kidney transplantation were retrospectively analyzed. First, according to the characteristics of post-onset patients and lung CT findings, broad-spectrum antibiotics and anti-fungal treatment were adopted, and subsequently targeted therapy, that is, withdrawal or adjustment of dosage and combination regimen of immunosuppressive agents, was employed depending on etiology, fungi and virus detection results. Among the 58 patients, 28 patients were injected methylprednisolone, and 30 patients took oral prednisone. Hyoxemia correction, support therapy and immune replacement therapy were applied.
RESULTS AND CONCLUSION:Thirty-nine of 58 patients (67.2%) were positive for pathogens, including 7 cases of simple bacterial pneumonia, 4 cases of fungal pneumonia, 3 cases of simple cytomegalovirus infection, and 25 cases of mixed infections (5 cases of multiple bacterial infections, 17 cases of fungal and bacterial co-infections, and 3 cases of fungi, bacteria and cytomegalovirus co-infections). Patients subjected to methylprednisolone treatment spent shorter time to recover their temperature than those undergoing oral prednisone (P<0.05). In addition, creatinine fluctuation range in the methylprednisolone group was less than that in the prednisone group (P<0.05). The results showed that intravenous injection of methylprednisolone may accelerate absorption of inflammatory exudate in the lung and shorten treatment time.

Objective To establish an effective PCR assay for detection of Bartonella, and application of this assay in tree shrew .Methods Sequence of Bartonella was obtained from NCBI Genbank .Three pairs of primers were designed based on this sequence .One pair of primers was determined through amplifying the major strains in China .Sixty tree shrew blood samples were tested with this PCR assay .The positive amplified fragments were sequenced to verify the reliability of this method .Results A PCR method for detection of Bartonella is successfully established , with a high specificity and the sensitivity was of 2.0 ×10 -5 μg/mL.Among the tested 60 blood samples , 15 positive cases were detected.Sequencing of the samples confirmed a 25%infection rate of Bartonella in the tree shrews, well consistent with the amplification results , and verified the applicability of this detection method .Conclusion The establishment of this method provides the basis for detection of Bartonella in tree shrew.

Objective To discuss the influence factors on the quality of Potassium dehydroandrograpolide succinate for injection,and to optimize its production process.Methods The related substances were taken as the estimated standards and the orthogonal experimental method for determining the best production process was adopted.Results The water temperature and pH value had a significant effect on the related substances.The best production process was:dispensing water temperature at 40℃,pH at 6.4 and filling time being 2 h.Conclusion The optimized production process was quality control and reasonable,which can be used in industrial production.

Objective To evaluate the biocompatibility of vessel extracellular matrix (VECM) with bladder smooth muscle cells of rabbits,and to discuss the feasibility of vessel extracellular matrix as a matrix for urinary tract reconstruction.Methods Primary cuhured bladder smooth muscle cells (RBSMCs) iso- lated from New Zealand rabbits were implanted on VECM (1?10~6 cells/ml).The effect of VECM on meta- bolic activity,attachment,proliferation of RBSMCs were monitored in vitro by inverted light microscopy and scanning electron microscopy.The extracts of VECM and emulsion were prepared as experimental group and positive controls separately.The culture medium was used as negative control,and simple culture medium without cells was used as blank control.The cell viability was monitored by MTT method after 1-,3-,5-d see- ding.The in vivo tissue response to VECM was investigated by implanting into the subcutaneous sites of the rabbits.Results VECM exhibited nontoxic and bioactive effect on RBSMCs.RBSMCs could be attached to and proliferated on VECM and remained their morphologies.The cell proliferation rates of experimental group were 95.61%、98.34%、102.91%,respectively,after 1,3,5 d;those of negative control group were 100.00% ,respectively;and those of positive control group were 35.14%、38.95%、32.66%,respectively. There was significant difference in the rate between experimental group and positive control (P＜0.01),and no significant difference in the rate between experimental group and negative control (P＞0.05).In vivo, VECM demonstrated favorable tissue compatibility without tissue necrosis and fibrosis.Conclusions VECM exhibits nontoxic and bioactive effects on primary cultured bladder smooth muscle cells.It is a suit- able material for urinary tract reconstruction.