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 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 .

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.

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

BACKGROUD: It is not consistent on whether melatonin correlates with the adolescent idiopathic scoliosis(AIS) demonstrated by the domestic and foreign scholars.OBJECTIVE: To study the pathogenesis of AIS and its correlation with melatonin.DESIGN: A nonrandomized age-matched controlled study screened with two tests.SETTING: The Second Department of Orthopaedics, Haikou People' s Hospital.PARTICIPANTS: The experiment was completed in the Second Department of Orthopaedics, Haikou People' s Hospital. Totally 8 198 in-school students.from over 10 schools of towns in Haifu District, 4 423 males and 3 775 females, aged 7 to 16 years were surveyed.METHODS: Forty-two adolescents who have been diagnosed with AIS were selected and 50 healthy age-matched adolescents were selected as controls. Melatonin was assayed with the radioimmunology and the data were processed statistically.MAIN OUTCOME MEASURES: The levels of serum melatonin in the two groups.RESULTS: The serum melatonin of the preadolescents below 10 years old in AIS group, especially of females, was less than that of adolescents in the control group, with significant differencein statistics.CONCLUSION: The serum melatonin correlates with pathogenesis of AIS.

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.

AIM: To investigate the time - effect relationship between the expression of rhodopsin and recoverin and photoreceptor damage induced by N - nethl - N -nitrosourea ( MNU) .
METHODS: Thirty-six 7-week old Sprague-Dawley ( SD ) rats were intraperitoneally injected with MNU ( 60mg/kg ) and were put to death by dislocation of cervical vertebra 6, 12, 24h; 3, 7d after injection ( 6 per group) , respectively. As a control, six rats were injected with phosphate buffer saline (PBS) 5mL/kg and sacrificed on d3 after injection. The degree of photoreceptor apoptosis was detected by HE staining, terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling ( TUNEL ) and transmission electron microscope ( TEM ) in the right eyes. The mRNA expressions of rhodopsin and recoverin were detected different time after injection by Western blot and immunohistochemical method in the left eyes.
RESULTS:The dissolution of photoreceptor nucleus and apoptosis body were first perceived at 12h by TEM; most of cells at outer nuclear layer were presented positive reaction. The apoptotic index reached peak ( 29. 7% ±2.3%) at 24h which was coincided with the observation of TEM. The results of immunohistochemistry displayed that rhodopsin and recoverin were on a declining curve with time extension. Furthermore, the results of Western blot indicated that rhodopsin had dramatic decline at 6h after injection (P<0. 05), and extremely significant difference comparing to control group after 12h ( P<0. 01 ); while recoverin dramatic declined at 12h, and extremely significant difference after 24h (P<0. 01).
CONCLUSION:60mg/kg MNU intraperitoneally injection one - time may specifically induce photoreceptor apoptosis, The mechanism of down - regulation of rhodopsin and recoverin may be related to the selected apoptosis of photoreceptors.

In temporomandibular disorders (TMD), pain takes place when neuropeptides stimulate synovial tissue to produce several cytokines such as interleukin (IL)-1β, IL-6 and tumor necrosis factor (TNF)-α, which activate neurons and glia of synovial membrane at the bilaminar regions of temporomandibular joint (TMJ). It has been reported that, after neurogenic differentiation, the synovial mesenchymal stem cells (SMSCs), deriving from TMJ, possess the same cytological features as the neuronal cells. This study examined the ability of substance P (SP) and calcitonin gene-related peptide (CGRP) to stimulate SMSCs and neurogenic SMSCs secreting inflammatory cytokines during TMD, evaluated the mutual effects of inflammatory cytokines and neuropeptides and tested the analgesic effect of hyaluronic acid (HA). The levels of IL-1β, IL-6 and TNF-α in SMSCs and neurogenic SMSCs in the presence of neuropeptides were measured by ELISA. SP and CGRP produced by SMSCs and neurogenic SMSCs were determined by RT-PCR and Western blotting. The results showed that the expression of SP and CGRP was significantly enhanced in the neurogenic SMSCs in response to IL-1β, IL-6 and TNF-α, and the effect was remarkably inhibited by HA. IL-1β, IL-6 and TNF-α, in return, could be enhanced in the neurogenic SMSCs upon stimulation by SP and CGRP. Neuropeptides and inflammatory cytokines might work mutually on the TMD pain. The HA-mediated analgesic effect may be implicated in the inhibition of SP and CGRP expression in neurogenic SMSCs.