Cloning and bioinformatics analysis of SBACT5 gene in Echinococcus multilocularis from Southern Qinghai Plateau / 医学研究生学报

Objective Sodium-bile acid cotransporter plays an important role in the development of Echinococcus.The present study aimed to clone sodium bile acid cotransporter gene in Echinococcus multilocularis (EmSBACT5) and to analyze the bioinformatics of its coding protein.Methods EmSBACT5 gene was amplified by reverse transcription RCR (RT-PCR) technology and its nucleotide sequence was sequenced.Bioinformatics softwares were used to predict and analyze the physical and chemical properties, hydrophilicity/hydrophobicity, transmembrane domain, post-translational modification sites, structural domain, secondary structure, tertiary structure, subcellular localization and biological functions of the coding protein.Results The complete open reading frame was amplified with 654 bp in length, encoding 217 amino acids.The homology of the nucleic acid sequence and amino acid sequence of EmSBACT5 gene were 98% and 96% with the published SBACT5 in Echinococcus granulosus (EgSBACT5) respectively.Protein analysis results showed that the molecular formula of EmSBACT5 protein was C1141H1797N273O284S11.Its relative molecular mass was 24240 and isoelectric point was 8.99.There were 9 post-translational modification sites and 4 typical domains.Alpha helical, β-sheet, β-turn and random coil accounted for 29.95%, 31.80%, 7.83% and 30.41%, respectively.This protein was a hydrophobic membrane protein and was mainly located in the cytoplasmic membrane, and it might play a role in the processes of material transport and signal transmission.Conclusion The EmSBACT5 gene was cloned successfully and the informatics characteristics of its coding protein were obtained, which provides basic information for prevention and control of echinococcosis.

Efficient identification of gene knockout mutant mediated by CRISPR/Cas9 by CELⅠ crude extracts / 生物工程学报

CRISPR/Cas9, emerged as an efficient and powerful gene editing technology, has become the mainstream genome editing technology. Constructing mutants using CRISPR/Cas9 system is of great significance to the functional study and breeding application of useful genes. As the basis of the technology, a method for identification of mutation with efficiency and lower cost is needed. In this report, we studied the factors influencing mutation detected by CEL Ⅰ crude extracts, such as the amount of protein, enzyme incubation time, PCR buffers. Under the optimized conditions, we can integrate the mutation detection steps into one-tube reaction. We used this system to examine the mutation types and frequency of rice stn1 mediated by CRISPR/Cas9. We also used this method to identify different mutation types including homozygous, heterozygous and bi-allelic mutations. The accuracy of this method reached 100% verified by sequencing. Altogether, our results showed that using CELⅠ crude extracts was an efficient and low cost method for identification of CRISPR/Cas9 mediated mutation.

Effect of different anesthetic techniques on perioperative changes in T-lymphocyte subsets in patients with cardia cancer / 中国医师进修杂志

Objective To observe the effects of different anesthetic techniques on T-lymphocyte subsets in patients with cardia cancer. Methods Thirty-two patients undergoing elective radical operation for cardia cancer who had been average assigned into two groups by random digits table, group Ⅰ :general anesthesia;group Ⅱ :epidural anesthesia combined with general anesthesia (16 cases in each group).Peripheral blood T-lymphocyte subsets were measured before induction, after anesthesia, end of operation, 1d,3 d after operation. Results CD3,CD4,CD8 and CD4/CD8 all decreased in two groups after anesthesia,end of operation and 1 d after operation than before induction (P < 0.05). The index almost returned to the baseline values at 3 d after operation in group Ⅱ [ (60.75 ± 4.22 )%, (39.65 ± 3.64)%, (25.90 ± 1.17 )%,1.57 ±0.15](P >0.05),while in group Ⅰ still lower [(55.83 ±5.20)%, (35.15 ±5.65)%, (23.00 ±1.03 )%, 1.47 ± 0.35 ](P< 0.05 ). The two groups ontrast had significant deviation at 3 d after operation (P<0.05). Conclusion Epidural anesthesia combined with general anesthesia can reduce depression of Tlymphocyte subsets induced by surgical trauma and anesthesia.

Management of chronic lateral ankle instability / 中华创伤杂志

Objective To retrospectively analyze management of chronic lateral ankle instability and evaluate its clinical results. Methods There were 74 patients including 43 males and 31 females at age range of 15-63 years (mean 39 years). All patients had at least half year of disease history, more than twice repeated strains and over six weeks of conservative management. Of all, 41 patients were trea-ted with surgical operations including modified Brostrom in 12 patients, Myerson in eight and Chrisman-Snook in 21. The aasoeiated pathological problems should be treated at the same time, ie, tenedesis for the peroneal tendon subluxations in six patients, Achilles tendon lengthening in nine, lateral shift of cal-caneal osteotomies in eight and clearance and holes-making for the osteochondral lesions in 13. Results Of all, functional instability was alleviated in 21 patients through conservative treatment before surgery. Of 53 patients with either functional or mechanical instability, 10 patients showed symptomatic relief, two refused surgical operation and the remained 41 patients were treated surgically. Thirty-nine patients trea-ted surgically and 28 treated conservatively were followed up for 6-91 months. The Roos Functional Out-come Score was used for validating the results. The average value of the ankle functional outcome was 86.24 and 97.34 respectively following operation and conservative management. Conclusions The full examination is needed to remove any possible pathological factors for the patients with chronic lateral ankle instability. The surgical operation can obtain satisfactory functional recovery of the ankle if the conserva-tive management is failed.

Effects of different anesthetic techniques on perioperative changes in T-lymphocyte subsets in patients with esophageal carcinoma / 中国基层医药

Objective To determine the effects of different anesthetic techniques on T-lymphocyte subsets in patients with esophageal carcinoma. Methods Forty patients were randomly assigned into general anesthesia group (group Ⅰ),or combined generae anesthesia with epidural anesthesia group(group Ⅱ). Peripheral blood CD3, CD4,CD8 were measured before induction ( T1 ), after anesthesia ( T2 ), end of operation ( T3 ), 1d ( T4 ), 3d ( T5 ) after surgery. Resolts CD3,CD4,CD8,CD4/CD8 decreased at T2 in the two groups. In group Ⅱ ,CD3,CD4,CD8,CD4/CD8 ratio almost returned to the baseline values at T4 ,while group Ⅰ did not. Conclusion Epidural anesthesia combined with general anesthesia can reduce depression of T-lymphocyte subsets induced by surgical trauma and anesthesia, and is the anesthetic tecnique of choice for cancer patients undergoing major operation.