AIM To study protective effect of [D-Ala~2, D-Leu~5]-enkephalin (DADLE) against hydrogen peroxide (H_2O_2) induced myocardial cell injury and its possible mechanisms. METHODS Myocardial cells were isolated from neonatal rats and cultured for 48 h. Then the cells were randomly assigned into normal control, H_2O_2（200 μmol·L~(-1)）, H_2O_2+DADLE(1 μmol·L~(-1)), H_2O_2+DADLE +naltrindole(10 μmol·L~(-1)) and H_2O_2+DADLE +U0126(10 nmol·L~(-1)) groups and cultured for another 48 h.[~3H]TdR incorporation assay and flow cytometry were used to measure the cell proliferation and apoptosis rate. The lactate dehydrogenase (LDH) activities in culture supernatant measured by using LDH activity kit. The superoxide dismutase (SOD) activity and malondialdehyde (MDA) content in cells were measured with xanthine oxidase method and color reaction of thiobarbituric acid, respectively. The expressions of extracellular signal-regulated kinase (ERK) and phosphorylated-ERK (p-ERK) were observed with Western blot. RESULTS ① Compared with normal control group, the incorporation of [~3H]TdR in myocardial cells of H_2O_2 group was significantly lower, apoptosis rate was higher, LDH activity and MDA content in cells were higher, while SOD activity in cells was lower. In addition, the ratio of IA_( p-ERK) /IA_( ERK) was decreased. ② Compared with H_2O_2 group, the incorporation of ［3H］TdR in H_2O_2+DADLE group was significantly higher, apoptosis rate was lower, LDH activity and MDA content in cells decreased, while SOD activity increased significantly. The ratio of IA_( p-ERK) /IA_( ERK) was increased. ③ δ-Opioid receptor antagonist naltrindole and ERK antagonist U0126 inhibited this effect of DADLE on the above index changes induced by H_2O_2. CONCLUSION The δ-opioid receptor has protective effect against H_2O_2-induced myocardial cell injury, and its possible mechanism may be related to its promotion of antioxide capacity and ERK phosphorylation.

Background and purpose: High-dose chemotherapy followed by autologous stem cell transplantation (auto-HSCT) is considered as the ifrst line treatment for patients with relapse/refractory lymphoma after conventional chemotherapy. However, most of these patients still relapse the second time. The purpose of this study was to analyze the efifcacy of the consolidation chemotherapy after autologous stem cell transplantation (HSCT) refractory/relapse lymphoma in high risk. Methods:A total of 38 patients with relapsed/refractory lymphoma including Hodgkin lymphoma (HL) and non-Hodgkin lymphoma (NHL) were included, who were underwent auto-HSCT in our transplan-tation department from Jan. 2010 to Dec. 2013. In treatment group, 19 patients received 2 courses of consolidation che-motherapy after auto-HSCT every 2 to 3 months, with the regimen of mini-BEAM or modiifed mini-CBV. Another 19 patients had no chemotherapy after auto-HSCT as control group. Results:The median follow-up duration was 17.2 and 7.5 months in the treatment and control group respectively. The follow-up data demonstrated prolonged progression-free survival (PFS) in the treatment group than the control group [24.7 months vs 7.8 months, P=0.029 under intend-to-treat analysis ITT;24.7 months vs 5.2 months, P=0.01 under per protocol analysis(pp)]. There is also a trend of improved overall survival (OS) in the treatment group (P=0.055, ITT). Conclusion:Consolidation chemotherapy after auto-HSCT for refractory/relapsed lymphoma patients delay the relapse and tend to improve the overall relapse rate.

Objective To discuss the measurement of bone tumor volume on the basis of three dimensional images segmentation technology. Methods Twenty patients with lacunar bone tumor from Tianjin hospital and Tongji hospital were included in the study from January 2010 to August 2010. There were 11 males and 9 females. Each patient was exposed to spiral CT preoperatively. Then these primitive CT dates were imported into digital orthopedics clinical research platform (SuperImage orthopedics edition 1.1, Cybermed Ltd). The volume and maximum diameter of bone tumor were measured before operation by three-dimensional reconstruction technology. The actual tumor volume was measured during the operation. The tumor volume was also calculated from plain X-rays and CT scans as ellipsoidal or cylindrical depending on the tumor configuration and presence or absence of a soft tissue component. Results The tumor volume was measured to be (14.92±7.34) mm3, (16.65±6.95) mm3 and (34.29±15.70) mm3 using three-dimensional reconstruction technology, intraoperative elevation, and traditional radiograph measurement separately. It was found that there was no difference regarding the outcomes of measurement between three-dimensional reconstruction technology and gross intraoperative measurement. But obvious difference was detected between gross intraoperative measurement and traditional radiograph measurement. Coefficient of correlation between diameter and volume of bone tumor was 0.325 (P=0.162). Conclusion Digital measurement is a precise, efficient,convenient and repeatable method for bone tumor measurement.

Objective Digital PCR ( dPCR ) was established to detect plasma epidermal growth factor receptor (EGFR) T790M mutation of non-small cell lung cancer (NSCLC) patients and was evaluated in terms of analytical performance and clinical application significance.Methods The specific primers and probes for EGFR T790M mutation and wildtype were designed to establish dPCR platform.Limit of blank, sensitivity and linearity of dPCR were evaluated by the detection of plasmids with different concentrations to set up optimal reporting system and reanalyzing process.The mutation of EGFR T790M in plasma and tissue samples from 10 patients with advanced NSCLC resistant to EGFR-TKI therapy who were enrolled in Zhongshan Hospital Fudan University from January 2014 to October 2015 were analyzed by dPCR and amplification refractory ( ARMS) , respectively.The consistency was evaluated between dPCR and ARMS by Chi-square test.The correlation of T790M abundance detected by dPCR between plasma and tissue samples was also analyzed by Peasrson correlation analysis.Results Limit of blank and sensitivity of dPCR was 10 copies and 0.01%, respectively.dPCR was evaluated as linear in the range of 0.01%-100%( Y=1.226X-3.984,R2 =0.999 ).The consistency between dPCR and ARMS of tissue samples was good ( kappa=0.80), while the positive rates of plasma T790M detected by dPCR was significantly higher than ARMS (50%vs 20%,P<0.05).It was found that T790M abundance detected by dPCR was highly correlated between lung cancer tissue and plasma ( R =0.923, P <0.05 ) using Pearson correlation analysis. Conclusions A new method of dPCR with high sensitivity and absolute quantification is established for the detection of EGFR T790M mutation in plasma from advanced NSCLC patients, which brings tumor liquid biopsy into real.It has the ability to provide the most direct and valuable guidance for clinicians to make decision on EGFR tyrosine kinase inhibitors therapy in patients with advanced NSCLC resistant to EGFR-TKI.

Using uniform design to optimize some relative factors influencing the expressing of t-PA variant-rPA(K) in E.coli system, and found the best expressive conditions. They were(by using 300ml culture fask): the volume of media was 25ml, inducing time was 5.3 hours, pH was 6.0, the concentration of IPTG was 0.1mmol/L, inducing time was 25℃, and best culture media was HD. After being optimized, the yield of expression had been improved from 0.16 to 0.48, and it was as 3 times as before. The results above will offer the basement for purification and renaturation of rPA(K).

The new intein-mediated PHB purify protein system is a high expression, automatic cutting, for purification, low-cost protein purification system,it is conducive to large-scale protein purification.Choose human antibacterial peptide LL-37 as the purification objects,which is poison to prokaryotic cell.We construct intein-mediated PHB purified human antimicrobial peptide LL-37 system through genetic engineering technology and use this system to purify LL-37. The results show that this system can highly express LL-37 fusion protein and purifiy the product as same size with expectations.

ObjectiveTo evaluate the clinical outcome of new proximal femoral nail antirotation (PFNA) in treatment of subtrochanteric fractures. MethodsThere were 31 patients with traumatic subtrochanteric fractures intramedullarily fixed with PFNA. The patients included 14 males and 17 fe-males at a mean age of 54.6 years (range 34-90 years). According to Seinsheimer classification of sub-trochanteric fractures, 14 patients were with type Ⅱ fractures (including four patients with type ⅡA,seven with type ⅡB and three with type ⅡC), 12 with type Ⅲ fractures (including eight with type ⅢA and four with type ⅢB), two with type Ⅳ fractures and three with type Ⅴ fractures. Close reduction was performed under traction and C-arm fluoroscopy in 26 patients and open reduction through a mini-incision before inserting the nail in five patients due to difficult close reduction. ResultsThe operation lasted for a mean time of 72.5 minutes (range 45-120 minutes), with mean blood loss of 127.5 ml (range 100-350 ml). Of all, 27 patients (87.1%, 27/31) were followed up for a mean duration of 14.3 months (range 8-24 months), which showed fracture union in all patients, with a mean union period of 17.4 weeks (range 10-21 weeks). According to Harris Hip Rate Scale, the results were. excellent in 21 pa-tients, good in two and fair in four, with excellence rate of 85.2% (23/27). ConclusionsPFNA is an effective device for treatment of subtrochanteric fracture, with a high union rate and a low complication rate, for it has advantages of easy use and minor trauma, low bleeding and stable fixation in operation.

Objective To analyze the related factors that influence the stone free rate ( SFR) in flexible ureteroscopic lithotripsy ( FURL ) and develop a stone free index ( SFI ) model to estimate and predict the outcome of FURL.Methods A total of 393 patients receiving FURL were included in this study from May 2013 to August 2014.All patients′and calculous characteristics were recorded.It was evaluated the correlation of one-stage SFR with body mass index, the degree of hydronephrosis, the sterile urine, the renal insufficiency, the stone location, the stone number, the cumulative stone diameter ( CSD) , the stone density, the average of CT values, the minimum angle of pelvis ureter long axis with lamp long axis, the average length of stone located calyx-neck, and the minimum ratio of stone located calyx-neck′width with calyx′width.Multivariate regression analysis was used to analyze the relationship between preoperative characteristics and the SFR.Results The one-stage postoperative SFR in our study was 92.4% ( 363/393).We found that the staghorn stone, bacteriuria, CSD, average of CT values, the average length of stone located calyx-neck, the minimum ratio of stone located calyx-neck′width with calyx′width were significantly correlated with the postoperative SFR ( P <0.05 ) .We used logistic regression analysis to determine statistical significant variables and to create predictable mathematical model.The SFI system was consist of four stone characteristics, including the staghorn stone, the cumulative stone diameter, the average length of stone located calyx-neck, and the minimum ratio of stone located calyx-neck′width with calyx′width.The SFI had a high ROC curve (AUC=0.867) for predicting the one-stage postoperative stone free outcome.SFI score >7.5 meant a relatively high SFR ( SFR>85%) of FURL.Conclusions A SFI model using preclinical data was developed to predict the postoperative outcome of FURL, as well as the one-stage SFR.This model needs further prospective studies in the future.

The complete genomic sequence of Duck hepatitis virus 1 (DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides (nt) in length with a 5'-terminal un-translated region (UTR) of 626 nt and a 3'-terminal UTR of 315 nt (not including the poly(A) tail). One large open reading frame (ORF) was found within the genome (nt 627 to 7 373) coding for a polypeptide of 2 249amino acids. Our data also showed that the poly (A) tail of DHV-1 has at least 22 A's. Sequence comparison revealed significant homology (from 91.9% to 95.7%) between the protein sequences of the virus in the Picornaviridae family, its genome showed some unique characteristics. DHV-1 contains 3copies of the 2A gene and only 1 copy of the 3B gene, and its 3'-NCR is longer than those of other picornaviruses. Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates (from 92.9% to 99.2%), indicating that DHV-1 is genetically stable.

Abstract: Objective　To analyze the detection of SARS‑CoV‑2 in household environment and public place environment of Huangpu District, and describe the feature of SARS‑CoV‑2 contamination in the environment exposure to the infected cases, so as to support the control strategies such as disinfection and health communication. Methods　The results of RT-PCR test for the environmental samples exposure to the cases infected by SARS‑CoV‑2 during February 1 to March 31 2022 in Huangpu District of Shanghai were collected as the research data. Pearson χ2 was used to test the significance of the differences between positive rates of SARS‑CoV‑2 contamination. Results　From February 1 to March 31, household environment samples had a higher positive rate (6.47%, 234/3 618) of SARS‑CoV‑2 contamination while the public place samples had a lower one (1.22%, 47/3 582) in Huangpu District of Shanghai (χ2=141.908, P<0.01). Among the household buildings, the lane houses of old style representing poorer living condition had the highest positive rates (8.31%, 96/1 155) of SARS‑CoV‑2 contamination while the apartments representing better living condition had the lowest (3.59%, 22/612) (F=5.25，P<0.05). Among the samples from household environment, samples regarding sewerage had the highest positive rates (13.30%,58/436) of SARS‑CoV‑2 contamination, while samples regarding the tool of cooking and sweeping had the lowest (3.10%,17/548) (F=9.84，P<0.01). Among the samples from public place environment, samples regarding entertainment tools had the highest positive rates (13.33%, 2/15) of SARS‑CoV‑2 contamination, while samples regarding the tool of cooking and sweeping had the lowest (0.62%, 4/646) (F=4.22，P<0.01). Conclusion　In the environment exposure to the SARS‑CoV‑2 infected cases, the disinfection, ventilation and cleaning should be intensified strictly. SARS‑CoV‑2's surviving in sewage environment should be evaluation dynamically. More health communication should be pushed to people of poorer living condition.