Objective To construct five shRNA-expression plasmids and to investigate the expression of Smad2 in TGF-?/ Smads signal transduction treated with shRNA-expression plasmid.Methods Five shRNA-Smad2 DNA sequences from mRNA sequence of mouse Smad2 gene were designed and synthesized.DNA oligonucleotides encoding an appropriate shRNA were inserted to shRNA expression vector respectively.Five shRNA-Smad2 expression plasmids were obtained and then transfected into NIH/3T3 cells.The suppressed expression of Smad2 was assessed by RT-PCR and Western-blotting.Results The shRNA-expression plasmid numbered 2.4 could markedly reduce the expression of Smad2.The suppression effect of the RNAi-pool composed of four different plasmids was more obvious than that of any single.Conclusion The shRNA-expression plasmids were successfully constructed,which could specifically and effectively suppress the expression of Smad2.The method of using a mixture of RNAi plasmids to improve the RNAi efficiency was established.

Objective It aims to investigate the relationships among the categories of Comprehensive Version for Stroke as described in the International Classiifcation of Functioning, Disability and Health (ICF) Core Set, and to provide new supports for Judicial Appraisal of functioning in stroke by ICF functioning mapping.Methods The variables of 59 categories of ICF assessment scale and the samples of 106 persons’ are selected and used in the least absolute shrinkage and selection operator (LASSO) for mining dependencies among those variables. The graphical modeling and analyzing with the software Gephi provides a visual map of the correlations among those classiifcations. Results 59 interconnected categories which organized into the functioning mapping. b340, b735, b175 and b152 are centrally positioned categories because of their high correlation.Conclusion Functioning mapping by graphical modeling can reveal complex relational structures embedded in functioning classiifcations, which provides the support for using ICF to appraisal stroke.

Objective To investigate the relationship between Ct value of mice liver and postmortem interval (PMI) under various ambient temperatures. Methods mice were stored at 10℃, 15℃, 20℃, 25℃ and 30℃ after execution, and total RNA was extracted from mice liver every 6 hours (PMI 6h to 72h). The levels of 18s rRNA were examined using real-time PCR. The results were expressed by cycle threshold (Ct) value to explore relationship between PMI and Ct value, and the interpolation functions were established to estimate PMI. Results In each group, Ct value increased with PMI increased. Surface equation was obtained after interpolation analysis on temperature range 10℃~30℃. The three-variable quintic surface equation was f(x, y)=-426.9+30.82x+44.48y-1.297x2-1.837xy-1.388y2+0.034 38x3+0.038 17x2y+0.038 67xy2+0.028 77y3-0.000 612 9x4-3.897e-7x3y-0.001 223x2y2+0.000 256 6xy3-0.000 537 4y4+3.606e-6x5-2.846e-6x4y+1.009e-5x3y2-3.439e-6x2y3-2.556e-7xy4+2.664e-6y5(r2=0.999 4). Conclusion The rule of Ct value changes at ambient temperature complied with three-variable quintic surface equation distribution. Measurement of interpolation function may be used for PMI estimation at ambient temperature.

OBJECTIVETo introduce the use of a self-made specimen protective sleeve in laparoscopic resection for upper or mid rectal cancer and sigmoid colon cancer with transrectal specimen extraction surgery and the improvement of implantation method, so as to avoid and reduce bacterial contamination and tumor cell dissemination in abdominal cavity.

METHODSDuring June 2015 and May 2017, 48 cases of high located rectal or sigmoid colon cancer were operated laparoscopically with natural orifices specimen extraction surgery (NOSES) using a self-made specimen protecting sleeve. Operation indication: (1) Rectum and sigmoid colon cancer with the distance of more than 6 cm from tumor inferior margin to dentate line. (2) The maximum diameter of intestine together with mesangial and tumor <7 cm by intraoperative judgment. (3) No anal and distal rectal surgery, no anorectal stenosis or lack of expansion capacity caused by trauma. (4) No ulcerative colitis, Crohn's disease or radiation proctitis. After transecting the rectum, the specimen protective sleeve was inserted through the right lower 12 mm main Trocar (This sleeve was tailored from the laparoscopic protective sleeve produced by China 3L Corporation, which was intercepted with 25-35 cm from one end of the sleeve according to the length of distal rectal retention. One end was ligated and the other was open with a ligature band. About 5 ml paraffin oil was used to rinse and lubricate during the operation). The rectal stump retained 7-8 cm in abdominal cavity. The transanal ligation part of the protective sleeve was cut off, then the stapler nail seat was inserted and specimen was pull out through the sleeve and rectum.

RESULTSThere were 30 males and 18 females. The average age was (64.5±14.1) years, the BMI was (25.4±3.9) kg/m, the tumor diameter was (3.3±1.1) cm, the maximum diameter of specimen was (5.4±1.5) cm and the length of specimen was (18.6±4.3) cm. Among these 48 cases, specimens of 36 patients were pulled out through inside of the sleeve easily, while specimens of 12 patients were quite difficult with resistance. Of 12 cases, 7 needed the help of transverse forceps, 4 needed to make 1 cm incision in pull-through bowel and insert a suction to decrease the volume of large specimens with gathering of gas and fluid, and 1 received small abdominal incision to remove specimen and perform intestinal reconstruction due to big specimen (the diameter of tumor and mesentery was 7.5 cm). Specimen tears of 6 patients didn't result in dissemination thanks to the specimen protecting sleeve. The operation time was (113.2±76.1) min, the bleeding amount was (38.5±17.3) ml, the time to first oral intake was (47.9±4.4) h, and the postoperative hospitalization length was (8.5±1.7) d. Anastomotic leakage occurred in 1 case (2.1%). No intra-abdominal and trocar infection, and obstruction were found.

CONCLUSIONThe use of protective sleeve and the improvement of the method of intraperitoneal implantation can effectively reduce the abdominal contamination during the specimen extraction. It can be applied to big specimens as well.

Objective The article is to study on the detection of α-helix proteins in post-traumatic epileptogenic focus by FTIR-mapping. Methods Fourier transform infrared spectroscopy-mapping were applied to identifying α-helix by point-by-point scanning in post-traumatic epileptogenic focus sections and to develop FTIR-mapping profiles. Result The high absorbance of α-helix is accord with post-traumatic epilepsy, there are some significant differences between high absorbance and low absorbance. Conclusion α-helix proteins are distributed in post-traumatic epileptogenic focus widely, thus α-helix protein are involved in post-traumatic epilepsy.

The article is comparative study about spine and extremities on clause for the identification of the body injured and The classification of the body impairment. We reviewed the terms and provisions about spine and extremities as follows, amputation impairment, function impairment, and amputation impairment combined function impairment. This paper provides a comprehensive access and analysisofthe similarities and differences between the two standards.

Objective To investigate the effects of changing the pancreatic stent number based on the last procedure on the prognosis of children and adolescents with recurrent chronic pancreatitis. Methods Data of 11 pediatric patients ( 3-16 years old) with chronic pancreatitis, undergoing ERCP for at least twice were retrospectively analyzed. Based on whether the number of stents changed in the second procedure, patients were divided into the control group( more than once and no change in the number) ,and the changing group ( more than once and change in the stent number in the second procedure ) . The hospitalization and surgery percentage and number of hospitalizations due to abdominal pain in follow-up period were compared to study whether number change of stents was efficient to cure pediatric patients with chronic pancreatitis. Results Eleven pediatric patients undergoing at least 2 ERCP procedures were recruited into our group. The hospitalization percentages were 140％( n=7) and 33. 3％( n=2) in the control group( n=5) and the changing group ( n=6) respectively( P<0. 001) . The number of hospitalizations due to abdominal pain was 1. 5 and 0. 8 in the control group and the changing group respectively ( P<0. 001) . The percentage of patient hospitalizations due to abdominal pain significantly reduced in the changing group ( P=0. 003) . Conclusion Increase in pancreatic stent number in the second procedure in pediatric patients can decrease hospital admissions related to the pancreatitis.

Objective:To explore the value of ex- vivo porcine stomach model for endoscopic submucosal dissection (ESD) training for international trainees. Methods:Fifteen international students received ESD training and learning for twenty days. Firstly, students learned basic theory of ESD and completed a questionnaire. Then they were randomized to receive endoscopic training either on the vitro animal (group A) or on training experience (group B) of the clinical observation. At last, one case was assessed by an experienced endoscopist. The total and step-by-step operating time, complete resection, size of specimen and complications were recorded. All students completed the questionnaire once again. SPSS 20.0 was used for t test and chi-square test. Results:There was significant difference in total ESD operation time between group A and group B ( P<0.05). There were significant differences between the two groups on cutting time and dissection time, but there was no difference on marking time, injection time and operating speed. There was only one case of block resection and perforation in each group. For group A, their mastery and clinic confidence of ESD had been obviously increased after the animal training course. Compared with the operation before the training in group A, it was found that the total time and cutting time of the ESD after the training were also significantly improved. Conclusion:Theory combined with endoscopic training on vitro animal model can make the trainees familiar with the basic theory and master the operational skills, which is helpful and valuable for them to perform ESD in further clinic practice.

OBJECTIVETo investigate the safety and feasibility of 3D laparoscopic surgery via transrectal extraction of specimens without abdominal incision in the treatment of slow transit constipation (STC).

METHODSFrom May 2015 to January 2017, 8 STC patients (6 females and 2 males) with informed consent were selected to receive subtotal colectomy with 3D laparoscopy as the no-incision incision group, in which the initial part of ascending colon and rectum were end-to-end anastomosed directly after extraction of the specimen through the rectum. Twelve STC patients (9 females and 3 males) undergoing traditional subtotal colectomy with 3D laparoscopy were selected as the traditional group by case matching method (gender, age, BMI, the difference of receiving operation time less than 12 months, same surgeon team). Perioperative parameters (operation duration, intraoperative blood loss, exhausting time, postoperative hospital stay, complications, postoperative pain score and additional pain management), inflammation index at postoperative day 1 and day 3 (leukocyte, procalcitonin, interleukin 6, C-reactive protein), postoperative peritoneal infection, wound healing, short-term and long-term efficacy, patient satisfaction evaluation (subjective hundred-mark system) at postoperative one year were compared between two groups.

RESULTSThere were no significant differences between two groups in operation duration, intraoperative blood loss, exhausting time, postoperative hospital stay and morbidity of complication (all P>0.05). Significantly lower pain scores at postoperative 6-hour (median 3.0 vs. 4.5, U=23.0, P=0.042), lower ratio of additional analgesic at postoperative day 1(1/8 vs. 7/12, P=0.040) were found in the no-incision group. Leukocyte level at postoperative day 1 was significantly lower in the no-incision group [(11.0±3.5)×10/L vs. (14.7±3.6)×10/L, t=-2.281, P=0.035]. C-reactive protein concentration at postoperative day 3 was not significantly different between two groups but with different trend [median 78.1(0.1 to 154.0) mg/L vs. 22.0 (7.0 to 55.9) mg/L,U=33.0, P=0.047]. There were no significant differences of interleukin-6 and procalcitonin between two groups(all P>0.05). All the patients had follow-up for 14-31 months. Subjective effectiveness score was 90±9 in the no-incision group and 94±6 in the traditional group without significant difference(t=-1.099, P=0.286). No long-term complications associated with abdominal infection was observed in the no-incision group.

CONCLUSION3D laparoscopic subtotal colectomy via transrectal extraction of specimens without abdominal incision in the treatment of STC has similar short-term and long-term efficacies compared with traditional laparoscopic assisted surgery, and does not increase the probability of abdominal contamination.

Colectomy ; methods ; Constipation ; surgery ; Female ; Humans ; Laparoscopy ; Length of Stay ; Male ; Operative Time ; Rectum ; Treatment Outcome