Objective To evaluate prognostic factors for early bronchopleural fistula after pneumonectomy with non small cell lung cancer,and establish a validated clinical model to estimate the risk of early-BPF.Methods We reviewed the medical records of 429 patients who underwent pneumonectomy for NSCLC at our institution.We used univariate and multivariate analysis to identify potential independent risk factors for early-BPF after pneumonectomy for NSCLC.A model to estimate risk of early-BPF was developed by combining independent risk factors.Results The rate of early-BPF after pneumonectomy for NSCLC was 6.5％ (28/429).Three factors were independently associated with early-BPF:neoadjuvant therapy (HR:2.406),bleeding (HR:2.171)and diabetes (HR:1.144).A scoring system for early-BPF was developed by assigning 2 points for each major risk factor (neoadjuvant therapy and bleeding) and 1 point for each minor risk factor(diabetes).Scores were grouped as low (0-1),intermediate (2-3),and high (3),yielding the rate of early-BPF was 14％,27％,and 43％,respectively.Conclusion This clinical model is established on the basis of independent risk factors.This model can be used as a predictive tool for early-BPF after pneumonectomy for NSCLC.

This paper introduces a programming cardiac trigger apparatus used for the study of quantitative measurement of myocardial blood flow with myocardial contrast echocardiography.The design of hardware circuit based on MCU and the scheme of software based on Keil C51 are mainly discussed.With stable and reliable working,this apparatus provides a kind of technical support for the study of quantitative measurement of myocardial blood flow with continuous intravenous injection of sonicated microbubbles.

Objective To explore whether dithiothreitol(DTT)is helpful for PreScission Protease to cut off the GST from GST?CT3 protein. Meth?ods The pGEX?6P?3/CT3 recombinant plasmid was transfected into Escherichia coli BL21,and the GST?CT3 fusion protein was purified by B?PER method. PreScission Protease was applied with 10 mmol/L DTT to cut off the GST,then the SDS?PAGE was performed for identification of the CT3 protein. Results Without DTT,it was very difficult for PreScission Protease to cut off the GST from GST?CT3 protein. However,in the pres?ence of 10 mmol/L DTT,PreScission Protease could cut off the GST easily as identified by SDS?PAGE. Conclusion 10 mmol/L DTT can help Pre?Scission Protease to cut GST from GST?CT3 protein,so as to achieve high concentration of CT3.

Objective To evaluate the anestheisa efficacy of periprostatic nerve block in transrectal ultrasound (TRUS) guided biopsy of the prostate.Methods A total of 223 patients received prostate biopsy in our hospital from July 2010 to December 2013 were retrospectively studied,and were divided randomly into two groups.One hundred and sixteen cases in nerve block group accepted local anesthesia of prostate capsule and periprostatic nerve block after local perineal skin anesthetia,and 107 cases in local anesthesia group only accepted local perineal skin anesthetia and local anesthesia of prostate capsule.Patients in the 2 groups underwent prostate biopsy successfully.The visual analogue scale (VAS) and complications were recorded.Results The age,serum PSA level before biopsy,prostate volume and the number of puncture needles had no significant differences between the 2 groups (P＞0.05).The average VAS score was 2.3± 1.1,and 4.9±2.3 in the 2 groups.The VAS had significant difference between the 2 groups (P＜0.05).The incidences of hematuria,hemospermia and urinary retention were 37.1％ (43/116),3.4％ (4/116) and 1.7％ (2/116) in nerve block group,and 39.3％ (42/107),4.7％ (5/107) and 1.9％ (2/107) in local anesthesia group.The difference was not significant (P＞0.05).Conclusion Periprostatic nerve block for TRUS guided biopsy of the prostate could be safe with good analgesic effect.

Aim To construct 3 D structure model of cardiac Cav1.2 channel and check its accuracy and re-liability.Methods Homology model of Cav1.2 chan-nel α1 subunit was constructed using SWISS-MODEL server.The model was submitted to an online testing server built by University of California and scored by it.The binding of Cav1.2 channel with blocker or drug was simulated by MOE software molecular docking pro-gram to check the model′s accuracy and reliability.Re-sults Both the target sequence Cav1.2 α1 C and the template sequence Cav1.1 α1 S searched by SWISS-MODEL server belonged to L-type Ca2+channel.Since the homology was 7 1.5% revealed by sequence align-ment,homology modeling was performed using automa-ted mode.L-type Ca2+ channel blockers Verapamil, Nifedipine and Diltiazem could bind to the 3 D structure model of Cav1.2 channel,while sodium channel bloc-ker TTX could not.Furthermore,active ingredient of traditional Chinese drug Praeruptorin A and Berberine could also bind to the 3D structure model of Cav1.2 channel.Conclusion The 3 D structure model of Cav1.2 channel was constructed successfully,which provides reliable materials for further studies and estab-lishes the foundation for the application of homology modeling in the study of 3 D structure prediction of ion channels.

Nano-scale particles of silk fibroin peptide (SFP) were prepared from discarded materials of cocoon or filature by dissolving and enzymolysis. Polyvinyl Alcohol films inlaid with silk fibroin peptide nano-scale particles (SFP in PVA) were prepared by blending nano-SFP and PVA in water according to different blending ratios. The films' characteristics and their promoting cell growth functions were investigated. Silk fibroin fiber was dissolved in 60% NaSCN solution, and was decomposed with alpha-Chymotrypsin, Trypsin and Neutral, respectively. The uniformity of size of SFP nano-particles prepared by Neutral was better and appeared about 80-150 nm. (SFP in PVA) films were characterized by infrared spectroscopy (IR) measurement which demonstrated the combination of SFP and PVA. Scanning electron microscopy revealed the PVA films already inlaid with SFP micro-segment. The surface and form stability in water of the (SFP in PVA) films with blending ratios of 10/90, 20/80, 30/70 and 40/60 were observed. And the results showed that SFP/PVA film with the blending ratio of 30/70 has smoother surface and better stability in water. The Chinese hamster ovary (CHO) cells were cultured, and the promoting cell growth function of (SFP in PVA) films was assessed by MTT colorimetric assay. These findings indicate that SFP/PVA (30/70) film has excellent function of promoting cell growth.
Animals ; CHO Cells ; cytology ; Cell Proliferation ; drug effects ; Cricetinae ; Cricetulus ; Fibroins ; chemistry ; Membranes, Artificial ; Nanoparticles ; chemistry ; Peptides ; chemistry ; Polyvinyl Alcohol ; chemistry

Objective To construct plasmid vectors of calmodulin(CaM)Mg2+binding site mutants,and to express,purify and identify the mutant proteins. Methods Three kinds of cDNAs coding for the mutated CaM were cloned into pGEX?6P?3 plasmid vectors. These recombinant plasmids were transfected into Escherichia coli BL21 to express GST fusion proteins of CaM mutants. The fusion proteins were purified with Glutathione?Sep?harose 4B beads and PreScission protease. Results Both enzyme digestion analysis and DNA sequence identification proved the successful con?struction of the CaM mutant plasmids. SDS?PAGE results showed the high purity of each CaM mutant protein. The concentrations of three CaM mu?tants were around 1.0 mg/mL. Conclusion Prokayotic expression vectors of CaM Mg2+binding site mutants were successfully developed,and the eli?gible CaM mutant proteins were obtained. This study provided an important basis for further study on CaM’s biological function.

Objective To observe the effects of nonylphenol(NP)on the intracellular calcium concentration changes and cell proliferation,and the involvement of GPR30 receptor in H9c2 cell. Methods The intracellular calcium concentration changes were recorded by using intracellular calcium determination method and cell proliferation was observed by MTT method in H9c2 cell. Results NP(1×10-10 mol/L)increased the intra?cellular calcium concentration changing amplitude and promoted the proliferation of H9c2 cells,while NP(1×10-6 mol/L)decreased intracellular calcium concentration changing amplitude and suppressed cell proliferation. G15 could block the promoting effect of 1×10-10 mol/L NP on the intracel?lular calcium concentration and cell proliferation,but could not block the inhibition of 1×10-6 mol/L NP on the intracellular calcium increase and cell proliferation. Conclusion The results indicate that NP affect rapid calcium signal changes and cell proliferation in non?monotonic dose dependent manner,and its mechanism may be due to the different involvement of GPR30 receptor in different concentrations.

Surgery for rectal cancer has obtained quick improvement in techniques and concepts in recent years but still has challenging areas. Colorectal surgeons always seek to make operations clearer and easier, so that surgery can be safer and less time-consuming while guaranteeing surgical goals. With this purpose, our team have explored to make innovations in operations for rectal cancer and translate relevant patents from 2009. We summarize our achievements in this article as follows: (1) Reverse Miles operation (perineal operation first then laparoscopic abdominal operation) with two relevant patents—specialized instruments bag for laparoscopic operations (patent number ZL201520442331.0) and accessory spotlight for ultrasound scalpel (patent number ZL20102 0137689.X). (2) Laparoscopic sphincter-saving surgery for low rectal cancer through marker meeting approach with two patents—vacuum rectal drainage tube with functions of irrigation and ventilation (patent number ZL201520374385.8) and sterile sleeve cover of ultrasound scalpel handle (patent number ZL201920648102.2). (3) Laparoscopic radical resection of colorectal cancer and natural orifice specimen extraction. Different methods were designed according to the location of the tumor that classified as 20-40 cm, 10-20 cm and 5-10 cm to anus. Two relevant patents were specialized instruments for natural orifice specimen extraction (patent application number ZL2017101480141) and plastic film sleeve for natural orifice specimen extraction (patent application number ZL 201921169857.0). Reformation of surgical technique and innovation of surgical instruments should be conducted by surgeons with innovative thinking who always seek the way to translate ideas to patents and then real products to promote surgical treatment.

Objective To construct a CaME141G fusion protein-expressing plasmid,and to express,purify,and identify the activity of the recombinant protein. Methods The 141st site of the wild type CaM,E (GAG),was mutated to G (GGG),using site-specific mutagenesis technology. Escherichia coli BL-21 was transformed with the mutant plasmid. The GST-CaME141G fusion protein was mass-cultured and induced for expression. Subsequently,the GST-CaME141G fusion protein was purified using GS-4B beads. PreScission protease was applied to remove the GST,the Bradford method used to determine the concentration of purified protein,and SDS-PAGE used to detect its relative molecular weight and purity. The GST pull-down assay was used to study the protein's biological activity. Results The CaME141G protein was successfully purified at a high concentration and purity. The protein could interact with PreIQ protein fragments from the myocardial CaV1. 2 calcium channel C terminal,in a CaME141G concentration-dependent manner. Therefore,CaME141G has the ability to bind with the CaV1. 2 calcium channel. Conclusion This study successfully constructed a CaME141G fusion protein-expressing plasmid and purified the CaME141G protein. This lays a foundation for regulating the function of CaM mutations in the myocardial CaV1. 2 calcium channel,and for the study of its relationship with diseases of the cardiovascular system.