Objective The purpose of the study was to investigate the changes and clinical significance of matrix metalloproteinase (MMP)-1,MMP-3,tissue inhibitor of metalloproteinase (TIMP),interleukin (IL)-1,tumor necrosis factor (TNF)-α and transforming growth factor(TGF)-β in juvenile idiopathic arthritis (JIA).Methods Thirty-two JIA subjects and 28 controls (traumatic arthritis patients) were included into this study.The MMP-1,MMP-3,TIMP,IL-1,TNF-α and TGF-β level in the serum and synovia were assessed by ELISA.The WBC count,the level of CRP,ESR,RF were also detected.Independent t-test and Pearson's analysis were adopted for data analysis.Results ① The level of MMP-1,MMP-3,IL-1 and TNF-α in the serum was (158±67) ng/ml,(212±89) ng/ml,(39±19) pg/ml,(26±10) pg/ml respectively,which was significantly higher than that of the control group (all P＜0.05) ; the ratio of MMP-3/TIMP-1 (0.86±0.32) was higher in the study group than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (248±88),(17±9) ng/ml respectively,which was lower than that of the control group (P＜0.05).The value of seral MMP-3,MMP-3/TIMP-1 was positively correlated with that of WBC,CRP,ESR in the JIA group (all P＜0.05).② The value of MMP-1,MMP-3,IL-1 and TNF-α in the synovia was (216±78) ng/nl,(766±291)ng/ml,(56±21) pg/ml,(36±14) pg/ml respectively,which was higher than that of the control group (all P＜0.05); the ratio of MMP-3/TlMP-1 (2.68±0.89) was higher than that of the control group (P＜0.05),while the value of TIMP-1,TGF-β was (286±88) ng/ml,(12±4) ng/ml respectively,which was lower than that of the control group (all P＜0.05).③ The value of MMP-1,MMP-3,TIMP-1,IL-1 and TNF-α in the synovia was higher than that in the serum (all P＜0.05),while the value of TGF-β was lower than that in the serum (P＜0.05).Conclusion The value of MMP-1,MMP-3,IL-1 and TNF-α increases both in the serum and synovia,while the value of TIMP-1 decreases.The value of TGF-β decreases,which may have protective effect on JIA.The ratio of MMP-3/TIMP-1 in the serum is positively correlated to inflammation parameters,which may be used to judge the activity of illness in JIA.

Background Rapamycin(RAPA)is a specific inhibitor of the mammalian target of rapamycin (mTOR).Researches showed that RAPA inhibits the proliferation of lens epithelium cells(LECs)and tumor cells and induces apoptosis of tumor cells.To investigate whether rapamycin has the inhibitory effect on retinal pigment epithelium(RPE)cells is very important for the prevention and management of proliferative vitreoretinopathy (PVR).Objective This study was to investigate the effects of RAPA on the proliferation and apoptosis of human RPE cells in vitro.Methods Human RPE cells(D407 strain)were cultured and passaged and then divided into regular culture group(blank control group),DMSO control group(0.1‰ DMSO +regular culture),and different concentrations RAPA-treatment groups(5,10,20,40,80,160,320 nmol/L).The proliferation(A490)of human RPE cells was detected using MTT,and the inhibitory rates of RAPA on the proliferation of RPE cells were calculated and compared among different groups at 12,24 and 48 hours.The apoptosis rates of the cells were analyzed among various groups by Hoechst staining after 12,24,48 hours.Results The inhibitory rates of RAPA on RPE cells were significantly different among various groups(F=484.451,P＜0.01)and evidently elevated in 20-320 nmol/L RAPA groups compared with DMSO control group(P ＜ 0.01).The inhibition of RAPA on the cells was considerably enhanced as the lapse of time(F=232.262,P＜0.01)with more dominant effects in 24 and 48 hours compared to 12 hours after addition of RAPA(P＜0.05-0.01).Compared with blank control group and DMSO control group,the apoptotic rates of the cells were evidently increased in 12,24,48 hours in 10 nmol/L RAPA group(all P＜0.05),and higher cellular apoptotic rates were found in 20-320 nmol/L RAPA groups(all P＜0.01).The alteration of cellular apoptotic rate showed a gradually incremental trend as the acting time of RAPA(F =625.584,P＜0.01).Karyorrhexis and mass-like density staining and chromatin substance were seen in RPE cells under the fluorescence microscope in ≥ 10 nmoL/L RAPA groups.Conclusions RAPA suppresses the proliferation and induces the apoptosis of human RPE cells in concentration-and time-dependent manner in vitro.

Background Normal ultrastructure is the anatomical basis of retinal pigment epithelial(RPE) cells to perform normal physiological function.At present the precipitation method is often used to detect the ultrastructure of RPE cells with transmission electron microscopy(TEM).Objective The aim of this study was to explore a simple and feasible approach to examine the ultrastructure of human embryonic stem cell-derived retinal pigment epithelial (hESC-RPE) cells.Methods hESCs were induced and differentiated into RPE cells by the spontaneous differentiation method,and the expressions of microphthalmia associated transcription factor MITF and paired-box gene 6 (PAX6),specific protein of RPE cells,in the cells were detected by immunofluorescence assay.hESC-RPE cells were inoculated into Transwell filter,and the ultrastructure of the cell sheet was examined under the TEM.Then the ultrastructure of the cell sheet specimens was compared with those of hESC-RPE cells from cell precipitation and RPE cell specimens of 90-day-old Long Evans rats.Results MITF and PAX6 were positively expressed in hESC-RPE cells.The normal ultrastructure were visible in the RPE cells of rats under the TEM,including apical microvilli,polarized melanin granules,cellular nucleus,basement membrane and intercellular junctions,and the ultrastructure of hESC-RPE cell sheet on Transwell was similar to the RPE cells in rats.However,only scatter melanin granules,nonpolar nucleus and scanty microvilli were observed under the TEM in the hESC-RPE cells by cell precipitation method.Conclusions Without digestion process,hESC-RPE cell sheet on Transwell can retain the normal ultrastructure of hESC-RPE cells under the TEM,with a more simple and reliable advantage.

Objective To study the effects of use of cyclamic acid on monitoring D-Dimer and predicting deep venous thrombosis on lower limbs during total hip arthroplasty.Methods Ninety-three cases patients who received total hip arthroplasty operations at Joint Surgery Department of the Central Hospital of Zhuzhou city and Zhuzhou Hospital Affiliated to Xiangya Medical College of Central South University from December 2015 to May 2016 were selected as subjects and randomly assigned into study group with 50 cases and control group with 43 cases.During the operations,cyclamic acid was used intravenously and locally as a routine in the study group while saline was utilized instead in the control group.The D-Dimer was dynamically monitored before operation and 1,3,5,7,9 d after the operation,and venous color ultrasonography of both lower limbs were taken 3,6,9 d after the operation to check the conformation of thrombosis.Results The total blood loss after treatment in the study group was (350.5 ± 65.2) ml,intraoperative blood loss of (129.3 ± 43.1) ml,postoperative drainage volume of (80.9± 12.6) ml,occult blood loss of (141.9± 20.6) ml,corresponding to the control group were (560.8±60.6) ml,(208.9± 57.8) ml,(150.8 ± 18.9) ml,(202.9±23.9) ml.The above indicators were lower in the study group than in the control group,the differences between the two groups were statistically significant(t =16.02,7.59,21.24,13.22,P＜0.05).There were significant differences in terms of D2-dimer level of 1,3,5 d after the operation between the study group and the control group (P＜0.05),but at 7,9 d after the operation,the difference between the two groups were not statistically significant(P＞0.05).The study group and the control group at 9 d after operation with color Doppler ultrasound examination showed that there were 1 cases of patients with calf vein thrombosis both in two groups,there was no significant difference between the two groups(2.00％ vs.2.32％,x2 =0.012,P＞ 0.05).Conclusion The proper use of cyclamic acid can reduce blood loss and will not increase the risk of thrombosis.Monitoring dynamically on D-Dimer and deep venous color ultrasonography on lower limbs is helpful for early detection of thrombosis after total hip arthroplasty.However,the use of cyclamic acid during total hip arthroplasty will affect the monitoring on D-Dimer and therefore needs to be taken seriously.

Objective To understand the changes and possible mechanism of the blood brain barrier(BBB) permeability induced by tumor necrosis factor(TNF-?) in vitro.Methods BBB model was established by coculturing allogenic brain microvessel end othelial cell(BMEC) and astrocyte(AS).The BBB model was divided randomly into normal control group,TNF-? group and Y-27632 pretreatment group.The changes of BBB permeability were evaluated by Gamma radioim muno assay counter.Results The Gamma radioimmuno assay indicated that the marker,~(125)I-BSA,across the BBB model in vitro was significantly increased after TNF-? treatment compared with control group,Y-27632 pretreatmen could prevent the permeability of BBB induced by TNF-?(P

Objective: To establish a multiplex PCR-microarray method for detecting important enteropahogens.Methods: Uniplex and multiplex PCR were performed to obtain the best primer sets for identifying the target bacteria at species and multi-species level.Fluorescent dyes were mixed into PCR reaction to determine whether it can affect the efficiency of amplification.To improve the efficiency of microarray,a 35 pairs primer-labeling system was optimized based on the hybridization results to find the best combination to avoid false negative results.Results: Specific PCR products were all obtained using species-specific primer sets.More preferential amplification may happen when more primer pairs were added to the reaction.The hybridization results showed a positive association between the efficiency of multiplex-PCR and signal intensity.Conventional PCR yielded more products than fluorescent dyes labeled PCR.Thirty-five primers were divided into three different combinations to label target respectively,hybridization results showed a high specificity.Conclusion: Mixing fluorescent dyes into PCR may reduce the efficiency of amplification and hybridization,but may have no effect on the analysis of hybridization results.The hybridization efficiency of microarray depends on the amplification efficiency of multiplex PCR.For microarray target labeling,three primer sets could be used to avoid negative hybridization led by preferential amplification of multiplex-PCR.It indicates that the multiplex PCR-microarray method is an attractive diagnosis tool for the high-throughput identification of enteropathogenic organisms especially for multiple causative agents and epidemiological investigations.

· ATM:To analyze bacterial spectrum and drug sensitivity in conjunctival sac of cataract patients of Kazak.
· METHODS:A total of 538 cases of conjunctival sac secretion in cataract patients of Kazak were collected.The samples were cultured and their sensibilities to antibiotics were tested.
· RESULTS: The bacterial culture was positive in 214 cases.The positive rate was 39.8%. The variety of pathogenic bacteria were mainly made up of gram positive cocci ( 88.3%), and most of them were Staphylococcus epidermidis ( 66.4%), followed by Micrococcus(9.8%).Sex had no effect on conjunctival bacteria rate in the cataract patients of Kazak, while age, place of residence had an effect on camier rate. The camier rate of conjunctival bacteria was significantly higher in people over 60 years old than that in people with age between 40 to 59 years old.And the people from city had a significant lower bacteria positive rate than those from countryside and pastoral. Most of grams were sensitive to Vancomycin, Teicoplanin, Rifampicin, Duly cloth mildew mutual and Amikacin, the tolerance was less than 20%, and they usually had higher tolerance to Penicillin, Erythromycin, Tetracycline and Chloramphenicol (>70%) .
·CONCLUSlON:Gram positivecocci is the most common bacteria in conjunctival sac in cataract patients of Kazak. Staphylococcus epidermidis was most common, followed by Micrococcus.The germ-carrying rate of conjunctival SAC in Kazakh population is associated with the patient’s age and area of residence.The clinical use of antibacterial drugs should be strictly grasp the indications, to reduce the incidence of bacterial resistance.

Cell Differentiation ; Gene Expression Regulation ; Liver ; cytology ; Stem Cells ; cytology

AIMTo investigate the effect of CSF contacting neurons (CSF-CNs) lesion in rat dorsal raphe nucleus (DRN) on the scores of morphine withdrawal symptoms precipitated by naloxone and the nNOS expression in dorsal horn of spinal cord, and study the relationship between the distal CSF-CNs in rat brain parenchyma and the development of morphine dependence and withdrawal.

METHODSChemical lesion of neurons the injection of cholera toxin subunit B with horseradish peroxidase (CB-HRP) into one of the rats lateral ventricles, TMBST reaction, nNOS immunohistochemistry and Western blot were used in this study.

RESULTSThe withdrawal symptoms by the naloxone precipitated attenuated obviously after the lesion of CSF-CNs in rat DRN, scores of all signs were significantly decreased about 38% compared to that of withdrawal group without lesion (P < 0.05). The withdrawal symptoms scores of vehicle withdrawal group and side lesion withdrawal group were not changed significantly (P > 0.05). Neurons in the location of CSF-CNs concentrated in the rat brain slices of lesion group were damaged obviously, there were only few CB-HRP positive neurons around the lesion location. But the location and the quantity of the CB-HRP positive neurons in the brain slices of the group without lesion was stable relatively, and their appearance was very clear. After the lesion, the nNOS expression and the quantity of the nNOS positive neurons in dorsal horn of spinal cord decreased significantly compared to that of withdrawal group without lesion (P < 0.05), but it also increased significantly compared to that of normal group and dependence group (P < 0.01).

CONCLUSIONThe lesion of distal CSF contacting neurons attenuated the scores of morphine withdrawal symptoms precipitated by naloxone and the nNOS expression in dorsal horn of spinal cord. The distal CSF contacting neurons in rat brain parenchyma partly participated in the development of morphine dependence and naloxone precipitated withdrawal possibly by the modulation of NO (nitric oxide).

Animals ; Brain ; drug effects ; pathology ; Male ; Morphine Dependence ; metabolism ; Neurons ; drug effects ; pathology ; Nitric Oxide Synthase Type I ; metabolism ; Raphe Nuclei ; cytology ; pathology ; Rats ; Rats, Sprague-Dawley ; Substance Withdrawal Syndrome ; metabolism