Objective:To establish a cell line stably expressing the transient receptor potential melastatin 2(TRPM2)channel for screening TRPM2 inhibitors based on PiggyBac transposition system.Methods:A plasmid PiggyBac-human TRPM2(pPB-hTRPM2)eukaryotic expression vector was constructed using PiggyBac transposition system.The plasmid and a helper plasmid were co-transfected into HEK293T cells to express TRPM2,which was identified by fluorescence and patch-clamp assays.The high throughput screening performance was assessed with the Z'factor.Calcium imaging and patch clamp techniques were employed to assess the initial activity of eleven compound molecules,confirming the inhibitory effects of the primary molecules on TRPM2.The protective effect of the screened compounds on damaged cells was validated using the oxygen-glucose deprivation/reperfusion(OGD/R)injury model and CCK-8 kit.The level of cellular reactive oxygen species(ROS)was detected by flow cytometry.The neuroprotective effects of the compounds were evaluated using a transient middle cerebral artery occlusion(tMCAO)mouse model.Results:The HEK293T cells transfected with pPB-hTRPM2-EGFP showed high TRPM2 expression.Puromycin-resistant cells,selected through screening,exhibited robust fluorescence.Whole-cell patch results revealed that induced cells displayed classical TRPM2 current characteristics comparable to the control group,showing no significant differences(P＞0.05).With a Z'factor of 0.5416 in calcium imaging,the model demonstrated suitability for high-throughput screening of TRPM2 inhibitors.Calcium imaging and electrophysiological experiments indicated that compound 6 significantly inhibited the TRPM2 channel.Further experiments showed that 1.0 μmol/L of compound 6 enhanced cell viability(P＜0.05)and reduced the level of ROS(P＜0.05)of SH-SY5Y under OGD/R injury.0.3 and 1.0 mg/kg of compound 6 reduced the cerebral infarction volume in tMCAO mice(both P＜0.05).Conclusions:A stable TRPM2 gene expressing cell line has been successfully established using PiggyBac gene editing in this study.TRPM2 channel inhibitors were screened through calcium imaging and patch clamp techniques,and an inhibitor compound 6 was identified.This compound can alleviate cell damage after OGD/R by reducing cellular ROS levels and has a protective effect against cerebral ischemia-reperfusion injury in mice.

Objective This study aims to evaluate the application of BOPPPS teaching model combined with case and problem-based learning(CPBL)and the scenario simulation in cardiovascular internal medicine education.Methods The study included cardiovascular internal medicine graduates from the 2022 and 2023 cohorts.The 2022 cohort(n=12)served as the control receiving traditional teaching methods,while the 2023 cohort(n=16)was the experimental group,which utilized the BOPPPS model combined with CPBL and scenario simulation.The students theoretical examination scores were assessed and a questionnaire survey was conducted to evaluate self-assessment of abilities and teaching satisfaction among the students in the experimental group.Results The examination scores of experimental group were significantly higher(97.94±3.46)as compared to that of control group(69.13±12.88)(P＜0.05).Over 93.75%of the students in the experimental group strongly or somewhat agreed that the combined teaching model stimulated their learning interest,improved learning efficiency,facilitated their capacity building of autonomous learning,theoretical knowledge,helped establish clinical reasoning,and improved their teamwork skills.Additionally,over 93.75%of the students were highly satisfied with the course design,emphasizing key content,pro-moting teacher-student interaction,and expressed willingness to continue with this teaching method in the future.Conclusions The application of the BOPPPS teaching model combined with CPBL and scenario simulation in cardio-vascular internal medicine education achieved excellent teaching outcomes.It effectively stimulated students interest in learning,enhanced autonomous learning abilities,and received high satisfaction ratings from students.

Acetaldehyde dehydrogenase 2 (ALDH2) is an important kind of aldehyde dehydrogenase in mitochondria, which has the function of eliminating acetaldehyde and other toxic aldehydes substances. Furthermore, it is abundant in liver and is closely related to the occurrence and development of a variety of liver diseases. ALDH2 genetic polymorphisms plays an important role in the occurrence of a variety of liver diseases in the human population.This paper mainly reviews the research progress of ALDH2 in liver diseases in recent years, with a view to provide theoretical basis for clinical prevention and treatment.

Objective:To develop the Exercise Knowledge and Behavior Questionnaire for Osteoporosis Patients suitable for China, and to test its reliability and validity.Methods:Guided by the theory of knowledge, attitude and practice, a questionnaire framework was constructed, and the first draft of the Exercise Knowledge and Behavior Questionnaire for Osteoporosis Patients was formed through literature review, group discussion, semi-structured interview, Delphi expert consultation and pre-investigation. From June to August 2020, convenience sampling was used to select 120 osteoporosis patients admitted to the First Affiliated Hospital of Soochow University as the research object to conduct a questionnaire survey to test the reliability and validity of the questionnaire. A total of 120 questionnaires were distributed, 110 valid questionnaires were recovered, and the valid recovery rate was 91.67% (110/120) .Results:Among 2 rounds of Delphi expert consultation, the expert authority coefficients were 0.880 and 0.900, respectively, and the Kendall harmony coefficients were 0.308 and 0.332, respectively, with statistically significant differences ( P<0.01) . The Exercise Knowledge and Behavior Questionnaire for Osteoporosis Patients included two sub-questionnaire of knowledge and behavior, with a total of 19 items, including 10 items in knowledge and 9 items in behavior. Exploratory factor analysis showed that 2 common factors were extracted from the exercise knowledge sub-questionnaire, and the cumulative variance contribution rate was 60.712%. A total of one common factor was extracted for the exercise behavior sub-questionnaire, and the cumulative variance contribution rate was 72.075%. The content validity index of the total questionnaire and each sub-questionnaire were 0.925, 0.921, and 0.929, respectively, and the item-level content validity index of was 0.857 to 1.000. The Cronbach's α coefficients of the total questionnaire and each sub-questionnaire t were 0.935, 0.830, and 0.953, respectively, and the split-half reliability coefficients of the total questionnaire and each sub-questionnaire were 0.979, 0.898, and 0.974, respectively, and the test-retest reliability coefficients of the total questionnaire and each sub-questionnaire were 0.878, 0.813, 0.860, respectively. Conclusions:The Exercise Knowledge and Behavior Questionnaire for Osteoporosis Patients has good reliability and validity, and can be used to evaluate exercise knowledge and behavior in patients with osteoporosis.

Objective To study effects of backpack gravity center position on kinetics and kinematics of lower-extremity joints in parachuting landing and evaluate the injuries. Methods Seven participants performed parachuting landing with backpack gravity center on three positions: low-back (position 1), upper-back (position 2) and abdomen (position 3). Results The peak vertical ground reaction force (GRF) with backpack on position 2 was significantly lower than that on position 1. The joint moment on sagittal plane of the hip with backpack on position 2 was significantly higher than that on position 1 and position 3. The joint energy absorption of the hip with backpack on position 2 was significantly higher than that on position 1. The angular displacement of the hip on sagittal plane with backpack on position 2 was significantly higher than that on position 1 and was significantly lower than that on position 3. The angular velocity of the hip on sagittal plane with backpack on position 2 was significantly lower than that on position 3. Conclusions Different positions of backpack gravity center could significantly influence kinetic and kinematic parameters of the hip. Backpack gravity center on upper-back position could decrease the lower-extremity injuries. The results can provide evidences for evaluating backpack gravity center and decreasing injuries in parachuting landing.

OBJECTIVE: To evaluate the efficacy of internal fixation of lateral and medial borders for displaced scapular body fractures via the minimally invasive approach. METHODS: The internal fixation of lateral and medial borders via minimally invasive approach was applied in surgical treatment of 23 patients with scapular body comminuted fractures from January 2014 to June 2018. The lateral approach was made straightly orienting over the lateral border of scapula. The dissection was taken down to the deltoid fascia. The deltoid was retracted cephalically, revealing the external rotators. Blunt dissection was used down to the lateral border between infraspinatus and teres minor, exposing the fracture site. The medial incision was done along the medial border of the scapula over site of the fracture. Dissections were taken down to the fascia and the periosteum. A subperiosteal dissection was then performed to elevate the infraspinatus to the degree necessary to visualize the fracture. The medial and lateral borders of scapula body were fixed with plates and screws in a frame-like way. RESULTS One patient developed the delayed healing of the incisions due to liquefactive fat necrosis. The other 22 patients showed no complications of the incisions. The glenopolar angle (GPA) of fractured scapula was increased from preoperative (25±12) degrees to postoperative (41±5) degrees (P<0.01). The healing time of fractures healed was 3-8 months, with an average time of (4.4±1.3) months. CONCLUSIONS The lateral-medial combined fixation through minimally invasive surgical approach for the scapula body fractures allows visualization of fracture reduction without extensive muscular or subcutaneous flaps, and is associated with successful fracture healing and high functional scores of the shoulder.

OBJECTIVE: To explore the causes of oocyte vitrification and its application in assisted reproduction. METHODS: We retrospectively analyzed the data of 26 patients with 27 cycles of oocyte vitrification cryopreservation undergoing intracytoplasmic sperm injection (ICSI) and embryo transfer between January, 2008 and October, 2018. The causes of oocyte vitrification and the outcomes of ICSI and clinical pregnancy were analyzed. RESULTS: The causes of oocytes vitrification included mainly azoospermia or severe spermatogenesis disorder of the husband, failure to obtain sperms from the husband, failure of the husband to be present on the day of oocyte retrieval and acute diseases of the husband to not allow sperm collection. A total of 274 oocytes were frozen in 27 oocyte retrieval cycles, and 217 eggs were thawed in 19 cycles with a survival rate of 81.11% (176/217). The normal fertilization rate, cleavage rate and high-quality embryo rate was 74.81% (98/131), 89.80% (88/98) and 36.73% (36/98), respectively. Fifteen patients underwent embryo transfer, and the clinical pregnancy rate and live birth rate was 53.33% (8/15) and 33.33% (5/15), respectively. Compared with patients below 35 years of age, the patients aged above 35 years had significantly lower oocyte survival rate after thawing (82.76% 74.42%, =0.211), clinical pregnancy rate (77.78% 16.67%, =0.041) and live birth rate (55.56% 0, =0.044). CONCLUSIONS Oocytes vitrification can be used as a remedy for infertile couples who fail to provide sperms due to male factors on the day of oocyte retrieval. Vitrification of the oocytes does not significantly affect the fertilization rate or the clinical pregnancy rate. The survival rate of the thawed oocytes is related to the age of the wife, and an age younger than 35 years can be optimal for achieving favorable clinical pregnancy outcomes after oocyte vitrification.
Adult ; Cryopreservation ; Embryo Transfer ; Female ; Humans ; Male ; Oocytes ; Pregnancy ; Pregnancy Rate ; Retrospective Studies ; Vitrification

Objective To explore the clinical effect of enhanced recovery after surgery and pain management during the perioperative period in rectal cancer patients.Methods 100 rectal cancer patients after radical resection were divided into ERAS group (50 cases) and routine care group (50 cases).Results Compare with the routine group,the time of ERAS group was shorter in postoperative bowel function recovery [(1.8 ± 0.6) d vs.(3.4 ± 0.6) d,t =-8.1,P ＜ 0.001],oral feeding [(1.3 ± 0.6) d vs.(3.2 ± 0.6) d,t =-10.1,P ＜ 0.001],intraperitoneal catheter drain [(3.6 ± 0.7) d vs.(5.3 ±0.8) d,t=-6.7,P＜0.001] and mobilization[(1.1 ±0.3)d vs.(2.7 ±0.5) d,t=-12.7,P＜0.001].ERAS group was associated with shorter hospital stay [(4.6 ± 0.6) d vs.(6.1 ± 0.6) d,t =-7.7,P ＜ 0.001],lower costs (P =0.014),lower pain score at the time of 6 h,12 h,24 h and 48 h after surgery (P ＜0.001).There was no significant statistical difference in postoperative complication rate 8％ and 10％ (P =1.000).Conclusions ERAS management in rectal cancer patients after radical operation enhanced postoperative recovery.

To screen the pathogenic mutation location in a genetic family with the neurofibromatosis (NF1) by the next generation sequencing and analyze the clinical phenotype,Illumina Miseq sequencing was applied to capture and analyze the target regions of NF1 family's probands,and furtherly find out the suspicious mutations,as well as to verify the family members by Sanger sequencing.Two rare variants were identified in proband,including the heterozygous missense mutation c.C3649T (p.P1217S) in KIF1B gene and the missense mutation c.T6311C (p.L2104P) on exon 41 of NF1 gene (NM_000267.3).The amino acid at position 2104 was found to be changed from leucine to proline in NF1.The protein prediction SIFT and Polyphen-2 values were 0,0.997,which predicted a conformational change in the encoded protein and eventually affected its function.The mutation c.T6311C in NF1 gene was detected in all patients in this family,which showed genetic co-segregation.The clinical phenotype was neurofibroma in the spinal canal.There were no café au lait spots,iris Lisch nodules,scoliosis,tinnitus,heating loss,or elevated intracranial pressure.The missense mutation c.T6311C (p.L2104P) in NF1 gene might be the genetic cause of this hereditary disease of neurofibromatosis.