Objective: To obtain monoclonal antibodies against programmed cell death 10(PDCD10) for further study of the structure and function of PDCD10 protein.Methods: Balb/c mice were immunized with recombinant PDCD10,hybridoma cell lines secreting monoclonal antibodies against PDCD10 were screened by regular cell fusion and subcloning approach.The specificities of these monoclonal antibodies were determined by ELISA,Western blotting and Immunofluorescecence assay.Results: Three hybridoma cell lines(5G1,4F7 and 3H5) stable in secreting specific monoclonal antibodies were successfully obtained.Subclass of IgG belonged to IgG1(4F7 and 5G1)and IgG2b(3H5),respectively.The ascite titers of these monoclonal antibodies reached 1∶10~7.They could specifically bind to recombinant PDCD10 and endogenous and overexpressed PDCD10 proteins proved by ELISA and Western blotting.They failed to react with E.coli lysates and glutathione S-transferase(GST).In addition,these three monoclonal antibodies could recognize different epitopes of PDCD10 proteins assessed by immune fluorescence competitive binding assay.Both endogenous and overexpressed PDCD10 protein mainly located in the nucleus.Conclusion: Monoclonal antibodies against PDCD10 with high titers and specificity have been successfully prepared,which has laid the foundation for further study of PDCD10 protein.

Objective:Mutations in epidermal growth factor receptor (EGFR) can predict tumor response to tyrosine kinase inhibitors (TKIs) in non-small cell lung cancer (NSCLC). However, not all cases of NSCLC with EGFR mutations can respond well;thus, discovering the heterogeneity of NSCLC at the molecular level is necessary. This study aimed to determine the discrepancy in EGFR mutations in primary tumors, N2 lymph nodes, and plasma samples. Methods:Primary tumors, N2 lymph nodes, and plasma samples obtained from 49 patients with stageⅢA-N2 NSCLC were analyzed for EGFR mutations in exons 19 and 21 by using mutant-enriched liquidchip technology. Results:In 49 patients, we detected 18 (36.7%) EGFR mutations in primary tumors, 11 (22.4%) mutations in N2 lymph nodes, and 2 (4.1%) mutations in plasma samples. Eleven (22.4%) cases showed discordance in EGFR mutations between primary tu-mors and N2 lymph nodes. In nine cases, EGFR mutations were detected only in primary tumors, whereas EGFR mutations were de-tected only in N2 lymph nodes in two cases. In addition, EGFR mutations were detected in the plasma samples of two patients, who al-so carry mutations in their primary tumors. Conclusion:A considerable proportion of NSCLC cases showed discrepancy in EGFR muta-tions between primary tumors and N2 lymph nodes. In addition, the detection rate of EGFR mutations was lower in plasma samples obtained from patients with stage IIIA-N2 NSCLC. All of the results indicated tumor heterogeneity at the molecular level during metas-tasis, and this heterogeneity may have implications during treatment with TKIs.

Objective To probe the therapeutic effect of ginger-salt-partitioned moxibustion on urge incontinence after stroke. Methods 40 stroke patients following urge incontinence were randomly divided into control group (n=20) and experimental group (n=20). 2 groups all received the same treatment, routine acupuncture and rehabilitation. Additionally, the experimental group received ginger-salt-partitioned moxibustion at Shenque (CV8). The curative effect was compared after treatment (4 weeks). Results 36 cases finished the treatment, 17 in the control group and 19 in the experimental group. Total number of urination, urinary incontinence and nocturia reduced, and the average volume of each urine increased (P<0.05), and volume of bladder residual urine reduced in 2 groups after treatment (P<0.001), while the experimental group was better than the control group (P<0.05). The scores of modified Barthel Index increased in 2 groups after treatment, but there was no significant difference between 2 groups (P>0.05). Conclusion The effect of ginger-salt-partitioned moxibustion at Shenque on poststroke following urination disorders is remarkable.

Objective To prepare recombinant adenovirus pAd-gal-9 containing murine galectin-9 and explore galectin-9's pro-apoptotic effect on T lymphocytes. Methods The recombinant adenovirus plas-mid pAd/CMV/V5-DEST-gal-9 was prepared by conventional molecular cloning and LR reaction. The pAd/ CMV/V5-DEST-gal-9 linearlized by Pac I was transfected into 293A cells with Lipofectin 2000. Eight days after transfection, the 293A cells were subjected to freeze/thraw circle for three times and the supernatant was collected after centrifugation. Higer titer pAd-gal-9 was produced by large-scale infection of 293A cells with the supernatant containing pAd-gal-9. The supernatant was condensed to get purified pAd-gal-9 by CsCl density gradient centrifugation. After titer determination with gradient dilution of harvested pAd-gal-9 infec-tion in 293A-seeded 96-wells, pAd-gal-9 was used to infect the CHO cell line. Immunohistological assay, Western blot and flow cytometry were employed to ascertain the subcellular location expression of galectin-9. We added solid-phase transgenic CHO cells or freshly-cultured supernatant to medium containing activated T cells to detect the pro-apoptotic effect of galectin-9. Results The pAd-gal-9 was prepared successful. Im-munohistochemical staining of CHO infected with pAd-gal-9 confirmed that galectin-9 was expressed in the cytosol. Intercellular staining indicated that mean fluorescence intensity of galectin-9 was significantly higher in pAd-gal-9-infected CHO group than control group. Supernatant from pAd-gal-9-infected CHO promoted the apoptosis of T cells. The percent of apoptotic T cells was higher than the Tim-3 positive T cells. Conclu-sion CHO infected with pAd-gal-9 can secret galectin-9 to promote the apoptosis of activated T cells via Tim-3-independent mechanisms.

Objective:To understand the views of respiratory nurses on the status quo of nursing quality evaluation in acute exacerbation of chronic obstructive pulmonary disease (AECOPD), and to preliminarily screen the key indexes, so as to provide reference for scientifically constructing the evaluation index system of nursing quality during AECOPD.Methods:Eighteen clinical nurses and nursing managers with rich experience and working in respiratory department from three general tertiary hospitals in Shanghai were selected by means of purpose sampling to conduct focus group interviews, and analyze the collected materials by means of Colaizzi content analysis.Results:Three themes were extracted, namely, lack of specificity and sensitivity of disease care in current nursing quality index, opinions and suggestions on the setting of AECOPD nursing quality index and the setting of key index of AECOPD nursing quality. Besides, twenty-five key indexes were obtained, inclusive of four structural indexes, sixteen process indexes and five result indexes.Conclusions:It is necessary and important to establish AECOPD nursing quality evaluation index system in a scientific manner and it is advised that the qualities of structure, process and result should all be taken into consideration in the course of establishment. The twenty-five key indexes picked out at this stage can serve as reference for further establishment of a scientific AECOPD nursing quality evaluation index system.

Objective To summarize the clinical characteristics and outcome of renal cyst infection in patients with autosomal dominant polycystic kidney disease (ADPKD). Methods Clinical data of 40 ADPKD patients with 43 episodes of renal cyst infection admitted in Shanghai Changzheng Hospital from 1st January 1991 to 31st December 2010 were retrospectively analyzed.Differences of microbiological data and treatments between 1st January 1991 to 31st December 2000 and 1st January 2001 to 31st December 2010 were compared. Results Among 473 identified patients with ADPKD and 662 episodes of hospitalization,40 patients had 43 episodes of renal cyst infection,including 8 definite and 35 likely cases.Microbiological documentation was available for 34 episodes (79.0％),Escherichia coli accounting for 82.4％ of all retrieved bacterial strains.Resistant Escherichia coli to quinolone and certain β-lactamine increased in recent decade.Clinical efficacy of initial antibiotic treatment was noted in 69.8％ of episodes. Antibiotic treatment modification was more frequently required for patients receiving initial monotherapy compared with those receiving combination therapy.In the first ten-year group,initial combination therapy and clinical efficacy were noted in 30.0％ and 60.0％ of episodes respectively,and hospital stay was (20.2±6.7) d.In the second ten-year group,initial combination therapy and clinical efficacy were noted in 61.9％ and 78.2％ of episodes respectively,and hospital stay was (16.3±3.2) d.Large infected cysts (diameter ＞5 cm) frequently required drainage. Conclusions In renal cyst infection,the source of the organisms is often a gram negative enteric organism.Empiric therapy is often initiated with two antibiotics.The drainage of large infected cysts remains the main treatment for cyst infection.

To identify acute renal allograft rejection biomarkers in human serum, two-dimensional differential in-gel electrophoresis (2-D DIGE) and reversed phase high-performance liquid chromatography (RP-HPLC) followed by electrospray ionization mass spectrometry (ESI-MS) were used. Serum samples from renal allograft patients and normal volunteers were divided into three groups: acute rejection (AR), stable renal function (SRF) and normal volunteer (N). Serum samples were firstly processed using Multiple Affinity Removal Column to selectively remove the highest abundance proteins. Differentially expressed proteins were analyzed using 2-D DIGE. These differential protein spots were excised, digested by trypsin, and identified by RP-HPLC-ESI/MS. Twenty-two differentially expressed proteins were identified in serum from AR group. These proteins included complement C9 precursor, apolipoprotein A-IV precursor, vitamin D-binding protein precursor, beta-2-glycoprotein 1 precursor, etc. Vitamin D-binding protein, one of these proteins, was confirmed by ELISA in the independent set of serum samples. In conclusion, the differentially expressed proteins as serum biomarker candidates may provide the basis of acute rejection noninvasive diagnosis. Confirmed vitamin D-binding protein may be one of serum biomarkers of acute rejection. Furthermore, it may provide great insights into understanding the mechanisms and potential treatment strategy of acute rejection.

Objective To explore the accuracy of endoscopic ultrasonography (EUS) for evaluating T3 esophageal squamous cell carcinoma (ESCC).Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 733 patients diagnosed with T3 ESCC by preoperative EUS who were admitted to the Sun Yat-sen University Cancer Center from January 2003 to December 2015 were collected.All the patients underwent radical resection of ESCC.The postoperative pathological stage as a gold standard,the accuracy,overstaged and understaged rates of clinical staging by preoperative EUS were assessed.Observation indicators:(1) comparison between clinical T staging evaluated by preoperative EUS and postoperative pathological T staging;(2) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect patients' diseases and postoperative survival up to December 30,2016.Overall survival time was from operation time to death or last effective follow-up.Measurement data with normal distribution were represented as (x)±s.Measurement data with skewed distribution were described as M (range).Count data were represented as cases and percentage.The survival curve was drawn by the Kaplan-Meier method,and survival analysis was done using the Log-rank test.Results (1) Comparison between clinical T staging evaluated by preoperative EUS and postoperative pathological T staging:all the 733 patients were confirmed as T3 ESCC by preoperative EUS.Postoperative pathological diagnosis showed that 9 patients were detected in pT1b,87 in pT2,630 in pT3 and 7 in pT4a.The accuracy,overstaged and understaged rates of preoperative EUS in evaluating T3 ESCC were 85.95％(630/733),13.10％(96/733) and 0.95％(7/733),respectively.N0,N1,N2 and N3 of postoperative pathological N stage were respectively detected in 329,247,110 and 47 patients.Twenty-seven,323 and 383 patients were in stage Ⅰ,Ⅱ and Ⅲ of TNM stage,respectively.The high-,moderate-and lowdifferentiated tumors were respectively detected in 125,403 and 205 patients.(2) Follow-up and survival situations:among 733 patients,639 were followed up for 1.0-153.0 months,with a median time of 29.0 months.The median survival time,1-,3-,5-year overall survival rates were 53.0 months (range,37.7-68.3 months),85.3％,58.1％ and 48.2％ in 733 patients,respectively.The 5-year overall survival rate was 75.2％ in 9 patients with pT1b,63.0％ in 87 patients with pT2,46.3％ in 630 patients with pT3 and 0 in 7 patients with pT4a,respectively,with a statistically significant difference (x2=24.089,P＜0.05).Conclusion There is a higher accuracy of EUS for evaluating T3 ESCC,however,the stage migration should be noted.

Objective To investigate the correlation between TYMS gene mRNA expression level and EGFR mutation in stage Ⅲ A-N2 non-small-cell lung cancer tissue (NSCLC).Methods The branched DNA-liquidchip technology (bDNA-LCT) and mutant-enriched liquidchip (MEL) technology were used to detect the TYMS mRNA expression and EGFR mutations at exon 19 and 21 in NSCLC tissues from 30 patients with stages Ⅲ A-N2 NSCLC,and the results were analyzed.Results Among 30 cases,low TYMS mRNA expression was in 14 cases (46.7 ％),middle to low TYMS mRNA expression in 7 cases (23.3 ％),middle mRNA expression in 7 cases (23.3 ％),middle to high TYMS mRNA expression in 0 case and high TYMS mRNA expression in 2 cases (6.7％);12 cases of EGFR mutation were detected out with the mutation rate of 40.0％,including 6 cases of exon 19 deletion and 6 cases of exon 21 missense mutation.The TYMS mRNA expression level was correlated with the EGFR mutation.The EGFR mutation commonly occurred in the tumor tissue of the patients with TYMS mRNA low expression (Z=-2.604,P=0.009).Conclusion The TYMS mRNA expression in NSCLC tissue is correlated with the EGFR gene mutation,which can provide references for the drug selection aiming at the patients with different conditions.

The etiology underlying ischemic stroke is still elusive. Previous studies have indicated that inflammation might play a key role in the pathogenesis, which provided a novel insight into the therapeutic strategy of ischemic stroke. In this review, we summarize some drugs which regulate the activation and polarization of microglia and further alleviate neurological symptoms.