Idiopathic Refractory Nephrotic Syndrome (IRNS) has been an intractable problem in clinical treatment .Anti CD20 monoclonal antibody is a new type of immunosuppressive agent , which can induce the lysis and apoptosis of B cells .Significant-ly, it improves the prognosis of IRNS patients .In recent years, a number of case studies and clinical trials have been conducted on the effectiveness of anti CD20 monoclonal antibody in the treatment of children with IRNS .In this paper, the mechanism, clinical applica-tion, adverse effects and problems in the study of anti CD 20 monoclonal antibody in IRNS will be reviewed .

Objective:To investigate the protective effect and potential mechanism of cordycepin on renal proximal tubular cells injury induced by lipopolysaccharide (LPS).Methods:Renal proximal tubular cells NRK-52E were incubated on a cell culture plated at a density of 1×10 5/mL for experiment, then divided into control group (Ctrl group), LPS group (cells were stimulated with 1 mg/L LPS), 10 μmol/L or 20 μmol/L cordycep in intervention groups (LPS+C 10 group and LPS+C 20 group). Cell viability was measured using cell counting kit-8 (CCK-8) reagent. The level of intracellular reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescin diacetate (DCFH-DA) staining. The protein expressions of inflammatory factors intercellular cell adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), interleukin-1β (IL-1β), and nuclear factor-κB (NF-κB) were detected by Western blotting. Results:Compared with the Ctrl group, LPS significantly inhibited NRK-52E cell viability, increased intracellular ROS, and up-regulated the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB. Compared with LPS group, after treated with 10 μmol/L or 20 μmol/L cordycepin, NRK-52E cell viability was significantly increased (Ctrl group as 1: 0.717±0.017, 0.916±0.036 vs. 0.554±0.046) and intracellular ROS level was significantly decreased (Ctrl group as 1: 1.527±0.165, 1.098±0.168 vs. 2.543±0.127), meanwhile the expressions of ICAM-1, VCAM-1, IL-1β and NF-κB were significantly down-regulated [Ctrl group as 1, ICAM-1/GAPDH: 2.364±0.097, 1.561±0.074 vs. 3.101±0.121; VCAM-1/GAPDH: 2.866±0.135, 1.920±0.098 vs. 4.170±0.119; IL-1β/GAPDH: 2.358±0.107, 1.563±0.179 vs. 3.301±0.210; phosphorylation NF-κB p65 (NF-κB p-p65)/GAPDH: 2.559±0.166, 1.596±0.148 vs. 3.183±0.098], the differences were statistically significant (all P < 0.05). Compared with the LPS+C 10 group, the cell activity of LPS+C 20 group was more significant (0.916±0.036 vs. 0.717±0.017, P < 0.01), and the expressions of ICAM-1, VCAM-1, IL-1β, NF-κB were down-regulated more significantly (ICAM-1/GAPDH: 1.561±0.074 vs. 2.364±0.097, VCAM-1/GAPDH: 1.920±0.098 vs. 2.866±0.135, IL-1β/GAPDH: 1.563±0.179 vs. 2.358±0.107, NF-κB p-p65/GAPDH: 1.596±0.148 vs. 2.559±0.166, all P < 0.05).Conclusion:Cordycepin could significantly increase the survival rate of NRK-52E cells, reduce intracellular ROS level, and inhibit inflammation, and the anti-inflammation effect can be related with NF-κB pathway.

Objective To evaluate puncture technique in thoracic endovascular aortic repair with abdominal aortic aneurysm and assess the feasibility and safety of using a pre-close technique for puncture and closure of femoral access sites. Methods From May 2010 to August 2013, the pre-close technique which involved two 6 F per-close ProGlide devices deployed in the femoral artery before upsizing to a 18-25 F sheath and one or two deployed before upsizing to a 14-16 F sheath were applied to 42 patients with abdominal aortic aneurysm (group A). Forty-seven patients using surgical femoral cutdown from December 2006 to April 2010 were enrolled into group B. The rate of technical success, time from procedure to the aortic delivery, operation time, low limb braking time, local complication, time from procedure to discharge, local vascular diameter after 3 months was evaluated and compared between two groups. Results There was no significant difference in endograft external diameter between two groups ( P>0.05). The rate of technical success was 97.62%(41/42) in group A and 95.74%(45/47) in group B, and there had no significant difference (P>0.05). Time from procedure to the aortic delivery, operation time and time from procedure to discharge in group A were significantly shorter than those in group B: (21.79 ± 5.79) min vs. (41.37 ± 11.79) min, (127.66±37.83) min vs. (157.84±42.71) min, (6.59±1.89) d vs. (9.14±2.57) d, P<0.05. The incidence rate of local complications, low limb braking time, and local vascular diameter after 3 months had no significant difference between two groups:7.14%(3/42) vs. 8.51%(4/47), (8.51± 1.83) h vs. (8.38±1.79) h, (1.05 ±0.36) mm vs. (0.98 ±0.31) mm, P>0.05. Conclusion The puncture technique with per-close ProGlide is safe and effective in percutaneous endovascular aortic repair which can be adopted as an alternative technique of surgical femoral cutdown approach in patients with abdominal aortic aneurysm.

Objective To investigate the correlations between the time of thoracic endovascular aortic repair (TEVAR) and prognosis in patients with type B acute aortic dissection (AADB). Methods The clinical data of 156 AADB patients with TEVAR was retrospectively analyzed and divided into 3 groups according to the time from onset of symptom to TEVAR:less than seven days was deifned as group 1 (G1, n=87), seven days to fourteen days group 2 (G2, n=48);more than fourteen days was group 3 (G3, n=21). The status of aortic reconstruction at three months TEVAR, in-hospital mortalities, mean hospital expense and length of stay were compared among three groups. Results Before TEVAR, there was no signiifcant differences in the ratio of smallest true lumen diameter and largest false lumen diameter amony the three groups (0.47±0.33, 0.42±0.18, 0.47±0.27, respectively, P>0.05). At three months after TEVAR, the ratio of largest true lumen diameter and largest false lumen diameter among the three groups was signiifcantly greater in group 1 (1.76±0.51) than group 2(1.42±0.30) and group 3(1.34±0.34, P < 0.05), when there was no signiifcant difference between the later two groups. Complete aortic reconstruction (8 from group 1 and 4 from group 2) was achieved in 12 patients at 3 months after TAVAR. Eight patients died during hospitalization, 5 from visceral ischemic, 2 from proximal aortic dissection, one patient from sudden death. Compared with G3, the hospital expense of group 1 and group 2 was cut down about ￥20000. Length of stay was signiifcant greater in group 3 than in group 1 and group 2 (P<0.05). Conclusions Early TEVAR for AADB was safe and beneifcial for aortic reconstruct and reducing the hospital expense and length of stay.

Objective To explore the regulation mechanism of autophagy-related protein, microtubule-associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines. Methods Human choriocarcinoma JEG-3 cell lines, and methotrexate resistant choriocarcinoma JEG-3 (JEG-3/MTXR) cell lines were used in our present study. Phosphorylation c-Jun (p-c-Jun) was evaluated after exposure to 0.02 ng/ml methotrexate for 72 hours in both cells by western blot. c-Jun gene was knockdown by small interference RNA (siRNA) in JEG-3/MTXR cells, and LC3 was evaluated by western blot and reverse transcription-PCR. The binding of LC3 promoter with c-Jun protein was detected via chromatin immunoprecipitation assay (ChIP) with or without 0.02 ng/ml methotrexate exposure. Results The results showed that p-c-Jun was up-regulated after methotrexate treatment for 72 hours (1.99±0.20, versus 0.20±0.06 at 0 hour;P<0.05) by western blot analysis in JEG-3/MTXR cell lines. Further investigation demonstrated that c-Jun-siRNA could inhibit the up-regulation of LC3 formation and after methotrexate exposure (LC3 mRNA:1.24±0.17 versus 3.03±0.43;LC3 protein:0.52±0.07 verus 1.20± 0.15; all P<0.05). The binding of LC3 promoter by c-Jun protein was up-regulated after methotrexate treatment by the method of ChIP in methotrexate resistant JEG-3/MTXR cells [(2.95 ± 0.35) times]. Conclusion Autophagy-related gene LC3 expression regulated by c-Jun protein may be involved in the effect mechanism of the development of methotrexate resistance in choriocarcinoma JEG-3 cells.

0.05).CONCLUSION:Scheme B has better compliance and is more economical as compared with scheme A.

Objective To investigate the clinical significance of thyroglobulin antibody (ATG) and thyroid peroxidase antibody (ATPO) in patients with vitiligo. Methods Venous blood samples were obtained from 87 patients with vitiligo and 90 age- and sex-matched normal human controls. Chemiluminescence was applied to measure the serum levels of ATG, ATPO, free triiodothyronine, free tetraiodothyronine and thyroid stimulating hormone (TSH). Results There was a significant increase in the positivity rates of ATG (23.0% vs 6.7%, P < 0.01) and ATPO (24.1% vs 7.8%, P < 0.01) as well as the serum level of TSH (3.4 ± 2.4 vs 2.4 ± 1.2 pmol/L, P < 0.05) in the patients with vitiligo compared with the normal human controls. It is worth mentioning that all patients positive for ATG or ATPO were diagnosed with vitiligo vulgaris. The positivity rates of ATG and ATPO in patients with vitiligo aged from 11 to 20 years and 21 to 40 years were significantly higher than those in age-matched normal controls (all P < 0.05). Also, female patients had a higher positivity rate of ATG and ATPO than female controls did (34.1% vs 8.5%, χ2 = 8.90, P < 0.01; 34.1% vs 10.6%,χ2 = 7.29, P < 0.05). The highest positivity rates of both ATG and ATPO were 53.3%, which were observed in vitiligo patients aged from 11 to 20 years, followed by patients from 21 to 40 years (ATG 34.5%, ATPO 34.5%). In patients with vitiligo positive for both ATG and ATPO, the occurrence of autoimmune thyroid disease was 70% (14/20), significantly higher than that in ATG- and ATPO- positive healthy controls (16.7%, χ2 = 5.4, P < 0.05). Conclusions ATG and ATPO were observed in young female patients with vitiligo vulgaris, and they may be associated with the development of autoimmune thyroid diseases.

Objective To study the gene mutations and clinical features of mandibuloacral dysplasia with type A lipodystrophy (MADA) in a Chinese family. Methods The information of 5 family members including 2 siblings suspected atyp-ical progeria was assembled. Genomic DNA was extracted from peripheral blood of 5 family members, the 12 exons of LMNA gene were ampliifed by PCR and then the PCR products were directly sequenced and analyzed by using Blast software online. The SIFT and PolyPhen-2 software were used to predict the harmfulness of mutations. Results The 2 siblings were clinically diagnosed as MADA. Heterozygous c.1579C>T (p.Arg527Cys) and c.1583C>T (p.Thr528Met) mutations were detected in this family. The father carried c.1583C>T (p.Thr528Met) mutation, the mother carried c.1579C>T (p.Arg527Cys) mutation, and their normal daughter were all heterozygous carriers with c.1583C>T (p.Thr528Met) mutation. Compound heterozygous c.1579C>T (p.Arg527Cys) and c.1583C>T (p.Thr528Met) mutations in 2 siblings led to MADA. The MADA showed an autosomal re-cessive inheritance pattern in this family. Conclusions The 2 siblings with MADA in this family were caused by compound heterozygous mutations in LMNA gene.

Objective To prepare monoclonal antibody(McAb) against human tissue kallikrein (HK) and develop an ELISA kit allows for the in vitro quantitative determination of human tissue kallikrein in urine. Methods To generate a monoclonal antibody specific for TK, the synthetic TK peptide consisting of 12 amine acids(12P), was fused to keyhole limpet hemocyanin(KLH) and used for immunization. Using hybridoma screening, monoclonal secreting cell lines were identified and used to generate ascites in BALB/c mouse. Antibody was purified by affinity column chromatography. 12% SDS-PAGE and Western blot were used to visualize the purified antibody. This kit employs indirect competitive ELISA technique and BiotinAvidin System. 12P was fused to bovine serum albumin(BSA) and has been pre-coated onto a microplate at first. Standards and samples were added to the appropriate microplate wells with a biotin-conjugated McAb croplate well. A TMB substrate solution is added to each well. The enzyme-substrate reaction is terminating by the addition of a sulphuric acid solution and the color change is measured spectrotometrically at a wavelength of 450 nm. The concentration of tissue kallikrein in the samples is then determined by comparing the O.D. of the samples to the standard curve. Results 8 hybridoma cell lines secreting mAbs special to HK,SDS-PAGE and Western blot demonstrated successful preparing and purification of McAb( 100% ). The linearity of this ELISA kit is demonstrated(r =0. 990). The range of detection of the assay is 0.008 μg/ml to 0. 5 μg/ml. The assay remained stable, with no change in the values measured, over five cycles of freezing and thawing. Conclusion 8 McAbs against HK have been prepared successfully and possess high titer and specificity. The development of an ELISA kit for detecting HK can meet the needs of detection of HK in urine samples.

Clinics are a main institutional form for doctors to open personal business in China .The develop-ment process of clinic reflects the situation of medical staff free practice .This study summarized the supervision poli-cies on clinic in China since the founding of China and got three conclusions .The first one was the attitude of the practice of the clinic has changed significantly .The change include four stages which were authorization ( 1949—1957 ) , limitations ( 1958—1977 ) , re-authorization ( 1978—1996 ) , promotion and encourage ( 1997—) along with macroeconomic system reform and the changes of government's governance ideas on health sector .The second conclu-sion was that the government gradually raised awareness of the status and role of the clinic institutions in the health system over the past several decades .The third conclusion was the supervision policies became more meticulous .In the future , clinical institutions can be a useful supplement to public medical institutions in China for its development process and characteristics .