Objective To study the endogenous metabolites of liver-kidney deficiency syndrome in knee-joint osteoarthritis (KOA) and explore the metabolic profile of KOA liver-kidney deficiency syndrome. Methods Totally 50 cases of KOA with liver-kidney deficiency syndrome and 50 cases of KOA with non-liver-kidney deficiency syndrome were collected respectively, and 20 cases of healthy volunteers were collected as the normal group. The serum samples of subjects were collected after fasting for 8 h. Hydro-Nuclear Magnetic Resonance (1H-NMR) spectrometer was collected. Principal component analysis and partial least squares discrimination analysis were used to conduct multivariate statistical analysis. Results 1H-NMR could identify 23 kinds of metabolites, and there was statistical significance between KOA patients and healthy volunteers (P<0.05, P<0.01). There was statistical significance in cartilage matrix metabolism, energy metabolism, lipid metabolism, pain, inflammation-related metabolites in patients with KOA liver-kidney deficiency syndrome and patients with KOA non-liver-kidney deficiency syndrome (P<0.05). Conclusion Patients with KOA liver-kidney deficiency syndrome have a unique 1H-NMR metabolic profile, KOA syndrome has a metabolic material basis.

Objective:To study the role of adhesion molecules in the pathogenesis of polymyositis. Methods:The abnormal expression of adhesion molecules on T cells in peripheral blood and muscle fibers from patients with myositis was analyzed by two colour immunofluoresence and RT PCR methods respectively. Results:The expression of adhesion molecules including lymphocyte function associated antigen 1(LFA 1 ),very late antigen 4(VLA 4) on T cells in peripheral blood and intercellular adhesion molecule l(ICAM 1) on muscle fibers from patients with myositis was markedly higher than that in the healthy control group. Conclusion: These findings suggested that adhesion molecules may be responsible for the migration of T cells and destraction of muscle fibers.

Aim ToexploretheeffectsofnewdrugT-006 on improving learning and memory abilities in scopolamine-induced dementia mice and its possible mechanism.Methods 72maleKunmingmicewere randomly divided into six groups:normal control group,model group,donepezil treatment group,T -006 treatment group with different doses(1,3 and 10 mg·kg-1 ).All mice were treated by intragastric ad-ministration for 14 consecutive days. Learning and memory abilities were tested by a five-day Morris water maze trial from the 1 1 th day.the first 4 days of the five-day Morris water maze,the navigation test was performed,the last day of Morris water maze is the spatial probe test.During the navigation test, mice were intraperitoneally given 2 mg · kg-1 scopolamine 20 minutes before entering the water,while normal control group mice administrated with sterile saline in-stead.Mice were not given T-006 nor scopolamine in spatial probe test.After Morris water maze,all mice were sacrificed for hippocampus and cortex.The activi-ties of AchE and SOD and the levels of GSH and MDA in hippocampus and cortex were measured after tissue harvesting.Results Comparedwithmodelgroup,T-006 could obviously improve learning and memory abil-ities in scopolamine-induced mice, significantly in-crease the levels of SOD and GSH and decrease the levelsofMDAandAchE.Conclusion T-006can significantly improve cognitive abilities in scopolamine-induced dementia mice,and its relevant mechanism may be closely related to its antioxidative effect and the ability to decrease AchE level.

AIM:To investigate whether gold nanoparticles (GNPs) reverses adriamycin (ADM), resistance of human hepatocellular carcinoma drug-resistant cell line HepG2/ADM and to explore the potential mechanism.METH-ODS:The sensitivities of HepG2 cells and HepG2/ADM cells to ADM were tested by MTT assay before and after GNPs pretreatment.The apoptotic rate was examined by flow cytometry.The concentration of ADM in HepG2/ADM or HepG2 cells was determined by ultraviolet-visible spectrophotometer .The content of glutathione ( GSH ) in HepG2/ADM or HepG2 cells by DTNB method.RESULTS:The half maximal inhibitory concentrations ( IC50 ) of ADM for HepG2/ADM cells were(29.46 ±1.73) mg/L and (15.18 ±0.85) mg/L before and after GNPs pretreatment,respectively.The IC50 of ADM for HepG2 cells was (9.16 ±2.03) mg/L before pretreatment.The apoptotic rate in GNPs+ADM group was higher than that in ADM group ( P<0.05 ) .The concentration of ADM in HepG2/ADM group was lower than that in HepG2 group (P<0.01).After GNPs pretreatment, the concentration of ADM in HepG2/ADM cells was higher than that before pretreatment.The content of GSH in HepG2/ADM group was higher than that in HepG2 group (P<0.01).After GNPs pretreatment, the content of GSH in the HepG2/ADM cells was lower than that before pretreatment.CONCLUSION:Gold nanoparticles can reverse ADM resistance of human hepatocellular carcinoma drug-resistant cell line HepG2/ADM,
reduce the content of GSH and increase the concentration of ADM in HepG2/ADM cells.

Objective To explore the changes in expression of Klotho, an aging-suppression protein, and neutrophil gelatinase associated lipocalin ( NGAL) and their relationship with rat mesangial cells ( RMCs) cultured with high glucose in vitro, and to explore the role played by Toll-like receptor-4 (TLR4) / nuclear factor-kB(NF-kB) p65 pathways in this process. Methods Three NGAL-siRNA sequences were designed and synthesized. The effective sequence in subsequent experiments was chosen. RMCs were preincubated with pyrrolidinedithiocarbamate (PDTC)or exogenously added Klotho prior to high glucose treatment. Realtime PCR was used to analyze Klotho, TLR4, NGAL mRNA expressions. Western blot was used to observe Klotho, TLR4,NF-kB p65, NGAL,fibronectin (FN), and connective tissue growth factor ( CTGF) protein expression. ELISA assay was used to detect monocyte chemoattractant protein-1 ( MCP-1) and CXCL5 secretions. Results High glucose suppressed Klotho expression significantly(P<0. 05) and activated TLR4 / NF-kB p65 pathway. Meanwhile,the levels of NGAL,FN,CTGF, MCP-1, and CXCL5 were highly expressed ( P < 0. 01). NGAL gene silencing obviously down-regulated the increased expressions of FN, CTGF, MCP-1, and CXCL5 ( P < 0. 01). After PDTC treatment the overexpression of NGAL protein was markedly lowered(P<0. 01). In addition, Klotho treatment significantly inhibited the activity of TLR4 /NF-kB p65 pathways and down-regulated the expressions of NGAL, FN, CTGF, MCP-1 and CXCL5 stimulated by high glucose(P<0. 01). Conclusion Klotho inhibits the activity of TLR4 / NF-kB p65 pathways and thus inhibits NGAL expression in RMCs cultured with high glucose in vitro. And then it suppresses the expressions of FN, CTGF, MCP-1, and CXCL5. This provides a new basis to illustrate the protection mechanism of the anti-aging protein Klotho in diabetic nephropathy, and may provide new ideas and therapeutic targets for prevention and treatment.

Objective To observe the results and reality of transurethral resection of bladder tumor(TUR-Bt) with upside down loop and plsamakinetic in the prevention of obturator nerve reflex.Methods A total of 211 patients with lumbar anesthesia received TUR-Bt.Firstly,the tumor tower above bladder wall was cut with the traditional way.When intraoperative findings were obturator nerve reflex,or to the obturator nerve mapping area,operation should be changed to superimpulse plasma column electrode vaporization to cut off residual tumor tissue,then after flipping the loop,the residual tumour was resected with upside-down loop from the tumour side wall.Results In 211 cases,192 cases had a little obturator nerve reflex,but the movement of body was not obvious,which did not influence the operation.The serious complication,such as perforation of bladder and injury of nerve and vessel,did not occur.Conclusions The lateral resection of bladder side wall tumour with upside-down loop can effectively avoid strong obturator nerve reflex,which is safe,uncomplicated,and efficacious and it does not increase the additional outlay and hurt.

Objective To study the radiosensitivity enhancement effects of recombinant adenovirus mediated retinoblastoma 94 gene on the growth of esophageal carcinoma cells EC109.Methods EC109cells was transfected with recombinant Rb94 gene adenovirus and irradiated by 137Cs γ-rays.The cohorts were divided into groups as blank control,Ad-LacZ,Ad-Rb94,radiation and Ad-Rb94 combined with radiation.Cell inhibition ratio,cell cycle and expression of retinoblastoma protein of EC109 cells were analyzed.Results The growth of EC109 cells transfected with Ad-Rb94,radiation and Ad-Rb94 combined with radiation group was all inhibited.The group of Ad-Rb94 combined with radiation resulted in greater inhibition of cells growth compared with Ad-Rb94 group and radiation group ( F =23.31,P ＜0.05 ).Cells of G2 phases of EC109 cancinoma cells for Ad-Rb94 combined with radiation group were the highest,which was 50％.The combination of Ad-Rb94 and radiation group resulted in the greatest expression of retinoblastoma protein,which reached 71％,significantly higher than Ad-Rb94 infection and radiation groups ( x2 =8.31,6.73,P ＜ 0.05 ).Conclusions Retinoblastoma 94 gene combined with ionizing radiation can enhance the radiation sensitivity of EC109 cells.

Objective:To study the application of HL-60 monocyte activation test in the pyrogen detection of vaccines. Methods:HL-60 cells were incubated with the standard pyrogen(endotoxin,zymosan and lipoteichoic acid)solution and the sample solution at different concentrations,and then IL-6 level was detected using ELISA kit. According to the concentration of standard pyrogen solution and the level of IL-6,the standard curve and linear relationship were obtained. Based on the IL-6 value from the sample solution,the pyrogen content of the sample was calculated. The standard pyrogen solution at middle concentration of the standard curve was used as the interference sample,and the interference assay was performed referring to the bacterial endotoxin test in Chinese pharmacopoeia. The pyrogen recovery rate and the content were detected using HL-60 / IL-6 assay in seven kinds of vaccines such as influenza vaccine(Split Virion),inactivated;hepatitis A(live)vaccine,freeze-dried;rabies vaccine(Vero Cell)for human use,freeze-dried;lyophilized human measles and rubella combined vaccine,live;measles and mumps combined vaccine, live; haemorrhagic fever with renal syndrome bivalent vaccine, inactived; group A and C meningococcal polysaccharide vaccine. Results:① The linear relationship between the pyrogen concentration and the IL-6 level was good,and R2 value was 0. 996,0. 993 and 0. 997,respectively. ② The recovery rate of pyrogen in the above vaccines ranged from 50% to 200% . ③ The endotoxin concentration of 10 batches of group A and C meningococcal polysaccharide vaccine was detected using HL-60 / IL-6 assay,and the average value of 6 batches from factory A was 134. 73 EU·ml-1 ,and that of 4 batches from factory B was 40. 75 EU·ml -1 . The pyrogen in the other vaccines was all negative. Conclusion:The HL-60 / IL-6 assay is suitable for the pyrogen detection of the seven kinds of vaccines.

The purity of 4,4′-dimethoxy-5,6,5′,6′-bis (methylenedioxy)-2′-morpholine methylenebiphenyl-2-methyl formate methanesulfonate (IMH), a new drug for fatty liver treatment, was determined through differential scanning calorimetry (DSC). Analysis of two-factor non repeatability method was performed in the investigation the effects of two factors (heating rate and sample weight) on purity determination. The DSC experimental parameters were optimized as follows: heating rate was 10 ℃·min^-1, temperature range was 150-300 ℃, sample weight was 2.0-4.1 mg, and N₂ flow rate was 80 mL·min^-1. The linear correlation coefficient (r) of this DSC method was 0.999 8. The RSD value (n = 6) of precision was 0.03%. The standard value and uncertainty of the purity results of the multiple batches of IMH drugs were (99.74 ± 0.29)%, (99.91 ± 0.28)%, (99.90 ± 0.28)%, and (99.81 ± 0.28)% with inclusion factor (K) of 2 and confidence probability (P) of 0.95. The results were basically consistent with the results of the mass balance method. The DSC mehod is a simple, rapid and accurate method, and provides a new reference method for determining the purity of IMH drugs, improves the accuracy and reliability of purity determination.

Primary Sj(o)gren' s Syndrome (pSS)is a chronic autoimmune disease with complex and diverse clinical manifestations.It is characterized by lymphocyte infiltration of exocrine glands such as the salivary gland and lacrimal gland leading to insufficient secretion of the gland,manifested as dry mouth and dry eyes.The disease in addition to exocrine glands cause damage,but also involving organs outside the gland,resulting in systemic systemic injury.The pathogenesis of pSS is not clear yet.Currently,symptomatic treatment is the mainstay of pSS,and the exact treatment is lacking.However,there are some new treatments in recent years.