Taking Naegleria australiensis, a species of pathogenic free-living amoeba (isolated in Shanghai in 1986), as antigen, two cell lines which provided potentially permanent source of monoclonal antibodies were established by lymphocytic hybridoma technique. The results of identification showed that: (1) the two cell lines could secret two different kinds of McAbs; (2) both of the McAbs were IgG (by gel diffusion); (3) McAbs produced in BALB/c mice were at high concentrations. One of them had a titer of ≥1 : 8 192 and the other≥l : 1 024 (by enzyme linked immunosorbent assay). A decline of the titers after purification by salting-out method was shown. One of the purified McAbs had a titer of≥1: 2 024 and the other≥l : 128.We have adopted two ways of recovering cryopreserved cells : ordinary recovering and "direct" recovering. The latter way was more practical because it could reduce the cycle of antibody production, and avoid contamination and chromosome variation. Experiments with different doses of cells revealed that, if the latter way was used, the optimal dose was 3 to 5 ? 106 cells per mouse.

Objective To observe the efficacy of combination therapy of calcium dobesilate dispersible and monosialotetrahexosylganlioside sodium on interleukin-6 (IL-6) and monocyte chemotactic protein-1 (MCP-1) in elderly patients with painful diabetic peripheral neuropathy.Methods From January 2012 to May 2017,in the Second People's Hospital of Lianyungang 70 patients of painful diabetic peripheral neuropathy,aged ≥60 years,were analyzed in this study.They were randomly divided into observation group (35 cases) and control group (35 cases).The observation group was treated with 40mg monosialotetrahexosylganlioside sodium dissolved in 250mL physiological saline,intravenous infusion per day,and oral calcium dobesilate dispersible 0.5g twice a day for two weeks.The control group was treated with methylcobalamin injection 0.5mg per day for two weeks.The clinical treatment effects and levels of IL-6 and MCP-1 were observed and compared between the two groups.Results After two weeks of treatment,the MDNS and MNSI scores of the observation group [(13.09 ± 5.38)points,(2.53 ± 1.19)points] were significantly lower than those of the control group [(18.31 ± 6.13) points,(4.19 ± 1.05) points,t =2.036,2.365,all P ＜ 0.05] and those before treatment [(21.26 ± 4.28) points,(5.40 ± 0.89) points,t =3.251,3.698,all P ＜ 0.05].The VAS-PI scores in the observation group [(6.24 ± 1.25) points,(4.13 ± 1.69) points] were significantly lower than those in the control group[(7.26 ± 1.28) points,(6.34 ± 2.65) points] at the first and second week (t =3.265,5.395,all P ＜ 0.05).The serum levels of inflammatory cytokines IL-6 and MCP-1 in the observation group [(15.16 ±0.88) ng/L,(157.19 ± 11.22) ng,/L] were significantly lower than those in the control group[(17.87 ± 1.19) ng/L,(198.21 ± 12.07)ng/L,t =2.152,1.365,all P ＜0.05]and those before treatment[(20.26 ± 1.05) ng/L,(260.44 ± 13.63) ng,/L,t =1.235,0.965,all P ＜ 0.05].Conclusion Combination of calcium dobesilate and mono-sialotetrahexosyl ganglioside may alleviate the sensory and pain sensations in patients with painful diabetic peripheral neuropathy,possibly by reducing the level of inflammatory cytokines IL-6 and MCP-1.

OBJECTIVETo make an open label prospective trial for comparing the therapeutic effects of mycophenolate mofetil (MMF) vs cyclophosphamide (CYC) pulse therapy on patients with diffuse proliferative lupus nephritis (DPLN).

METHODSForty-six patients with biopsy proven active DPLN were enrolled in this study. Twenty-three patients were given MMF orally at a dosage of 1.0 - 1.5 g/d (MMF Group). Another 23 cases received conventional intermittent CYC pulse therapy (CYC Group). Supplemental steroid treatment was offered in the same manner to both groups. The age, sex distribution and severity of renal damage were matched in two groups. Therapeutic effects were evaluated at the end of six-month treatment. Fifteen patients in the MMF Group and 12 patients in the CYC Group had repeated renal biopsy at that time.

RESULTSMMF therapy was more effective in reducing proteinuria and hematuria. A 50% reduction of urinary protein and urinary red blood cell excretion from baseline value in 69.6% and 91.3% patients in the MMF Group, while only 47.8% and 65.2% in the CYC Group. MMF was more effective in inhibiting autoantibody production (especially anti-dsDNA antibody) and in decreasing serum cryoglobulin levels. Pathologically, the MMF group showed more markedly reduction in glomerular immune deposits with less glomerular necrosis, and less microthrombi, less crescent formation and vascular changes in the repeated renal biopsy as compared with the CYC group. Adverse reactions related to the treatment included gastrointestinal symptoms 26.1% and 43.5% in the MMF and CYC Groups respectively, infection 17.4% in the MMF group and 30.4% in the CYC group.

CONCLUSIONMMF was more effective in controlling the clinical activity of DPLN and renal vascular lesions as compared with CYC pulse therapy in a 6 month follow-up study.

Adult ; Cyclophosphamide ; adverse effects ; therapeutic use ; Female ; Gastrointestinal Diseases ; chemically induced ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Infection ; chemically induced ; Kidney ; drug effects ; pathology ; Lupus Nephritis ; drug therapy ; Male ; Mycophenolic Acid ; adverse effects ; analogs & derivatives ; therapeutic use ; Pneumonia ; chemically induced ; Prospective Studies ; Treatment Outcome

The objective was to investigate the hypoglycemic action of catalpol in spontaneous diabetes db/db mice. 40 db/db mice were randomly divided into fi ve groups: model control gourp; db/db plus catalpol 40, 80, 120 mg/kg body wt. groups and db/db plus metformin 250 mg/kg group. Age-matched db/m mice were selected as normal control group. The mice were administered with corresponding drugs or solvent by gavage for 4 weeks. The oral glucose tolerance test was carried out at the end of 3rd week. After 4 weeks of treatment, the concentrations of fasting blood glucose (FBG), glycated serum protein (GSP), insulin (INS), triglyceride (TG), total cholesterol (TC) and adiponection (APN) in serum were detected. The protein expressions of phosphorylation-AMPKalpha1/2 in liver, phosphorylation-AMPKalpha1/2 and glucose transporter-4 (GLUT-4) in skeletal muscle and adipose tissues were detected by western blot. Real time RT-PCR was used to detect the mRNA expressions of acetyl-CoA carboxylase (ACC) and Hydroxymethyl glutaric acid acyl CoA reductase (HMGCR) in liver. Our results showed that catalpol could significantly improve the insulin resistance, decrease the serum concentrations of INS, GSP, TG, and TC. The concentrations of APN in serum, the protein expression of phosphorylation-AMPKalpha1/2 in liver, phosphorylation-AMPKalpha1/2 and GLUT-4 in peripheral tissue were increased. Catalpol could also down regulate the mRNA expressions of ACC and HMGCR in liver. In conclusion, catalpol ameliorates diabetes in db/db mice. It has benefi t eff ects against lipid/glucose metabolism disorder and insulin resistance. The mechanism may be related to up-regulating the expression of phosphorylation-AMPKalpha1/2.
Acetyl-CoA Carboxylase ; Acyl Coenzyme A ; AMP-Activated Protein Kinases ; Animals ; Blood Glucose ; Blotting, Western ; Cholesterol ; Fasting ; Glucose ; Glucose Tolerance Test ; Insulin ; Insulin Resistance ; Liver ; Metabolism ; Metformin ; Mice* ; Muscle, Skeletal ; Oxidoreductases ; RNA, Messenger ; Triglycerides

Objective To investigate the relationship between plasma tau protein, phosphorylated tau protein (p-tau) protein and cognitive function in subjects with generalized brain atrophy. Methods A total of 100 subjects with moderate and severe brain atrophy were divided into two groups according to cognitive function: normal group (n=50 cases) and dementia group (n=50 cases). And their gender, age, educational level, etc. are recorded. The tau protein and p-tau protein content in plasma were measured by double antibody sandwich enzyme-linked immunosorbent assay (ELISA). The differences between plasm tau and p-tau protein expression and their relationship with cognitive function were analyzed. Results Plasma tau protein and p-tau protein levels were significantly higher (P<0.05) in the dementia group [(210.92±43.79)pg/mL、(81.15±16.85)pg/mL] than in the normal group[(210.92±43.79)pg/mL、(81.15±16.85)pg/mL]. Plasma tau protein and p-tau protein levels were negatively correlated with the MMSE score (P<0.05) and had no significant correlation with the degree of brain atrophy (P>0.05). Conclusion Cognitive impairment may be associated with elevated tau protein levels in patients with extensive brain atrophy.