1.Significance and successful construction of stable transfection of MH7A cells with lower TRAF2 expression using lentiviral vector
Luying CHEN ; Liping JIANG ; Weikang WANG ; Shujun ZUO ; Jiajie KUAI ; Yang MA ; Chenchen HAN ; Wei WEI
Acta Universitatis Medicinalis Anhui 2024;59(2):193-199
Objective To construct a stable synovial cell line MH7A from rheumatoid arthritis(RA)patients using lentiviral vectors that interfere with the expression of tumor necrosis factor receptor associated factor 2(TRAF2),and to study the role of TNF-α-TRAF2 signaling in MH7A abnormal proliferation.Methods Based on the design principles of human TRAF2 gene sequence and shRNA sequence,three pairs of TRAF2 shRNA interference se-quences were designed and synthesized.The primers were annealed by PCR,and a linear vector was obtained by double enzyme digestion PLKO.1-puro.The linearized vector was connected to the annealed primers through Solu-tion I,and the connected products were introduced into receptive cells.The plates were coated,and positive colo-nies were selected for sequencing.Three different recombinant plasmids of PLKO.1-TRAF2-shRNA lentivirus were constructed,and lentivirus packaging plasmids was used to package logarithmic growth phase HEK 293T cells.Vi-rus solution was collected to infect MH7A cells.At the same time,puromycin was used to screen MH7A stable transgenic strains with low TRAF2 expression.CCK-8 method,Western blot,and qPCR were used to detect the proliferation function of MH7A induced by TNF-α and low expression of TRAF2,as well as downstream signal TRAF2,P65 protein expression and mRNA levels.Results PLKO.1-TRAF2-shRNA(1),PLKO.1-TRAF2-shR-NA(2),and PLKO.1-TRAF2-shRNA(3)lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were successfully constructed.The three TRAF2-shRNA lentivirus vector plasmids and control group lentivirus vector plasmids PLKO.1-puro were respectively introduced into the lentivirus packaging plasmid of HEK 293T to obtain virus solution.After infecting MH7A cells with the virus solution,they were treated with puromycin(2.00 μ G/mL)screening and obtaining MH7A stable transgenic plants after 2 days.Through qPCR and Western blot results,it was found that the expression of TRAF2 mRNA and protein in PLKO.1-TRAF2-shRNA(1)MH7A stably transfected cells was significantly reduced compared to the negative control group.The results of CCK-8 and Western blot showed that after knocking down TRAF2 in MH7A,the proliferation of MH7A cells with low TRAF2 expression induced by TNF-α and the phosphorylation level of P65 were significantly reduced.Conclusion A sta-ble transgenic strain of PLKO.1-TRAF2-shRNA(1)MH7A cells was successfully constructed to investigate the role of TNF-α-TRAF2 signal activation in mediating abnormal proliferation of RA synovial cells.
2.Effects of body mass index on plantar pressure
Chenchen ZHU ; Yuan YIN ; Yingji ZHOU ; Tingting MA ; Liyao SU ; Ming ZHANG
Chinese Journal of Tissue Engineering Research 2024;28(18):2808-2813
BACKGROUND:Human plantar pressure can reflect the health status of the lower limbs and even the whole body,which is an important basis for gait analysis,and body mass index is an important influencing factor. OBJECTIVE:To investigate the effect of body mass index on plantar pressure. METHODS:Twenty young college students from Xuzhou Medical University,including 10 males and 10 females aged 19-21 years,were selected as test subjects and divided into three groups according to the body mass index value:overweight group(body mass index>25 kg/m2,n=3),lean group(body mass index<18 kg/m2,n=4),and normal group(body mass index 18-25 kg/m2,n=13).A natural walking gait test was carried out on the three groups of subjects with a Zebris pressure distribution measurement plate to obtain the complete gait cycle parameters.The time proportion of support time phase,peak pressure,time to peak force,peak force and impulse volume were analyzed and the correlation between each parameter and the body mass index was analyzed by Person analysis. RESULTS AND CONCLUSION:(1)Compared with the other two groups,the time proportion of support time phase of subjects in the overweight group was relatively small,while the time proportion in the foot heel contact period and forefoot extension period was relatively large.There was a positive correlation of the time proportion of the foot heel contact period and forefoot extension period with body mass index,while there was a negative correlation between the time proportion of the arch support period and body mass index.(2)The peak pressure of the left arch and palm of the foot of the subjects of the overweight group was higher than that of the normal group,and the peak pressure of the left and right palm of the foot of the lean group was lower than that of the normal group.The peak pressure was positively correlated with the body mass index during the foot heel contact period.There was a significant positive correlation between the peak pressure of the left foot and body mass index during the arch support period as well as the peak pressure of both feet and body mass index during the forefoot extension period.(3)Plantar peak force time in the order of the gait cycle in increasing order:heel
3.Idioms reflecting the interplay between cognition and emotion in cognitive behavioral therapy
Jingren XU ; Hua YANG ; Chenchen LIN ; Na LIU ; Hui MA ; Tao PEI ; Mingyu HU ; Ning ZHANG
Sichuan Mental Health 2024;37(4):360-363
Background Cognitive behavioral therapy(CBT)is a widely used and empirically supported psychotherapy method.In recent years,the localization of CBT related to culture has received increasing attention from Chinese scholars.However,there is still a lack of systematic research on the localization of CBT using idioms as materials.Objective To explore whether idioms reflect the mutual influence between the two core therapeutic elements of CBT(cognition and emotion)and to use them as CBT treatment tools among college students,so as to provide references for the localization of CBT.Methods Using the Xinhua Idiom Dictionary(second edition)published by the Commercial Press,a three-round manual screening method was used to select idioms that reflected the mutual influence between cognition and emotion in CBT.Based on the selected idioms,a survey questionnaire titled"survey on idiom understanding level"was developed.From August to October,2022,stratified sampling was conducted among college students from different majors and universities,and 815 valid questionnaires were obtained through the Questionnaire Star platform.By analyzing the understanding of each idiom among college students,idioms that could be used as CBT treatment tools were identified.Results In the Xinhua Idiom Dictionary(second edition),there were 13 idioms that reflected the influence of cognition on emotion and 5 idioms that reflected the influence of emotion on cognition.12 idioms could be used as CBT treatment tools,including 8 idioms that reflected the influence of cognition on emotion and 4 idioms that reflected the influence of emotion on cognition.Conclusion There may be idioms that reflect the mutual influence between the two core therapeutic elements of CBT(cognition and emotion)and these idioms can be attempted as a therapeutic tool for CBT among college students.
4.Potential of new self-crosslinked hyaluronic acid gel on the recovery of endometrium after artificial abortion: a multicenter, prospective randomized controlled trial
Chunying LI ; Lirong TENG ; Qing LIN ; Liping ZHAO ; Yunxia ZHU ; Xin MI ; Zhenna WANG ; Xiaoye WANG ; Lisong ZHANG ; Dan HAN ; Lili MA ; Wenpei BAI ; Jianmei WANG ; Jun NI ; Huiping SHEN ; Qinfang CHEN ; Hongmei XU ; Chenchen REN ; Jing JIANG ; Guanyuan LIU ; Ping PENG ; Xinyan LIU
Chinese Journal of Obstetrics and Gynecology 2024;59(11):864-870
Objective:To evaluate the impact of self-crosslinked hyaluronic acid (SCH) gel on endometrium recovery after artificial abortion.Methods:A multicenter, prospective randomized controlled trial was conducted across 18 hospitals from December 2021 to February 2023, involving 382 women who underwent artificial abortion. Participants were randomly allocated to receive either treatment with SCH gel (SCH group) or no treatment (control group) in a 1∶1 ratio. The primary outcome was endometrium thickness in 14 to 18 days after the first postoperative menstruation. Secondary outcomes included changes in menstrual volume during the first postoperative menstruation, menstruation resumption within 6 postoperative weeks, time to menstruation resumption, duration of the first postoperative menstruation, and incidence of dysmenorrhea.Results:Baseline characteristics of participants were comparable between the two groups (all P>0.05), with 95.3% (182/191) in SCH group and 92.7% (177/191) in the control group completed the study. The postoperative endometrial thickness in SCH group was significantly greater than that in the control group [(9.78±3.15) vs (8.95±2.32) mm; P=0.005]. SCH group also had significantly fewer participants with reduced menstrual volume [23 cases (12.6%, 23/182) vs 31 cases (17.5%, 31/177); P=0.038]. Although SCH group experienced less dysmenorrhea during the first postoperative menstrual period, this difference was not statistically significant [28.5% (51/179) vs 37.1% (65/175); P=0.083]. Outcomes were similar between SCH group and the control group regarding the proportion of participants who resumed menstruation within 6 weeks postoperatively, time to menstruation resumption, and duration of the first postoperative menstruation ( P=0.792, 0.485, and 0.254, respectively). No serious adverse events were observed during the study period, and no adverse events were attributed to SCH gel treatment. Conclusion:The application of SCH gel after artificial abortion is safe and might aid in the recovery of the endometrium.
5.PKM1 Regulates the Expression of Autophagy and Neuroendocrine Markers in Small Cell Lung Cancer
TANG CHENCHEN ; JIN YULONG ; ZHAO PEIYAN ; TIAN LIN ; LI HUI ; YANG CHANGLIANG ; ZHONG RUI ; LIU JINGJING ; MA LIXIA ; CHENG YING
Chinese Journal of Lung Cancer 2024;27(9):645-653
Background and objective Small cell lung cancer(SCLC)is known as recalcitrant cancer with high malignancy and heterogeneity.Immunotherapy has changed the treatment pattern of extensive-disease SCLC(ED-SCLC),but the beneficiary population is limited.Therefore,exploring new therapeutic strategies is an urgent clinical problem to be solved for SCLC.SCLC is characterized by highly active glycolytic metabolism and pyruvate kinase Ml(PKM1)is one of the isozymes of PK,an important rate-limiting enzyme in glycolysis pathway.Previous studies have shown that PKM1 is related to autophagy and drug sensitivity,however,how PKM1 regulates drug sensitivity in SCLC and its mechanism remain unclear.The aim of this study was to investigate the biological functions of PKM1 in SCLC,including its effects on proliferation,migra-tion,autophagy,drug sensitivity,and expression of neuroendocrine(NE)-related markers in SCLC.Methods Western blot was used to detect the expression level of PKM1 in SCLC cells.PKM1 gene-overexpressed SCLC cell lines were constructed by stable lentivirus transfection.Proliferation of cells and drug sensitivity were detected by MTT,and migration ability of cells was determined by Transwell.The level of autophagy was detected by flow cytometry.Western blot was used to determine the expression levels of NE-related proteins.Results PKM1 was differentially expressed among various SCLC cell lines,and was lower in H1092 cells(P<0.01).Compared with the control group,there was no significant difference in proliferation level of PKM1 overexpressing H1092 cell,but the migration ability was significantly increased(P<0.001),the drug sensitivity was re-duced,and the level of autophagy was inhibited(P<0.001).Additionally,overexpression of PKM1 could upregulate the expres-sion of non-neuroendocrine(non-NE)-related proteins(P<0.01)and decrease the expression of NE-related proteins(P<0.01).Conclusion PKM1 was differentially expressed in SCLC cell lines,and high expression of PKM1 did not affect the prolifera-tion,but affected the migration of SCLC cells.PKM1 might affect drug sensitivity by inhibiting autophagy and regulating the expression of NE markers.These results provide a theoretical basis for exploring the role of PKM1 in SCLC.
6.Tislelizumab bridging to umbilical cord blood transplantation therapy for relapsed/refractory acute myeloid leukemia: report of 1 case and review of literature
Chenchen LIU ; Shiyuan ZHOU ; Qian ZHU ; Chao MA ; Xiao MA ; Depei WU ; Xiaojin WU
Journal of Leukemia & Lymphoma 2023;32(3):161-165
Objective:To explore the efficacy of tislelizumab combined with umbilical cord blood transplantation (UCBT) in relapsed/refractory acute myeloid leukemia (R/R AML) patients.Methods:The diagnosis and treatment of 1 patient with R/R AML who received tislelizumab bridging to UCBT after the failure of re-induction treatment in the First Affiliated Hospital of Soochow University in November 2021 was retrospectively analyzed.Results:The 59-year-old male patient with R/R AML achieved a complete remission after initial induction chemotherapy regimen of decitabine and venetoclax, and then additional consolidation therapy regimens of decitabine and middle-dose cytarabine, middle-dose cytarabine and idarubicin were performed. The patient relapsed 16 months later and failed to achieve a second remission after re-induction therapy regimens of cladribine, azacitidine, venetoclax combined with chemotherapy, and homoharringtonine, cytarabine combined with granulocyte colony-stimulating factor. Tislelizumab significantly reduced tumor burden and the patient achieved the complete remission after bridging to UCBT. After transplantation, the patient was given maintenance treatment with azacitidine and he had sustained remission without severe transplant-related complications during 9-month follow-up.Conclusions:The use of tislelizumab bridging UCBT can be a potential therapeutic strategy for R/R AML patients.
7.Early neurological deterioration in patients with minor ischemic stroke caused by large vessel occlusion: risk factors and the impact of rescue endovascular thromboectomy on outcomes at discharge
Chenchen MA ; Mengmeng WANG ; Jian DING ; Ting WU ; Shiying SHENG ; Hanqing LI ; Meng LIU
International Journal of Cerebrovascular Diseases 2023;31(5):321-326
Objective:To investigate the risk factors of early neurological deterioration (END) in patients with minor ischemic stroke caused by large vessel occlusion (LVO) and the impact of rescue endovascular thromboectomy (REVT) on clinical outcomes of patients with END at discharge.Methods:Consecutive patients with acute minor ischemic stroke caused by LVO within 24 h of onset in the Third Affiliated Hospital, Soochow University from January 2021 to March 2023 were retrospectively enrolled. Minor ischemic stroke was defined as baseline National Institute of Health Stroke Scale (NIHSS) score ≤5 at admission. END was defined as an increase of ≥4 in the NIHSS score within 24 h after the best medical management. The modified Rankin Scale was used to evaluate the clinical outcomes of patients with END at discharge. 0-2 was defined as a good outcome. Multivariate logistic regression analysis was used to identify the independent risk factors for END and the impact of REVT on clinical outcomes in patients with END. Results:A total of 75 patients with minor ischemic stroke caused by LVO were included, of which 31 (41.3%) developed END and 13 (41.9%) underwent REVT after END. Multivariate logistic regression analysis showed that internal carotid artery occlusion was an independent risk factor for END (odds ratio 4.304, 95% confidence interval 1.213-15.270; P=0.024), and REVT was an independent protective factor for good outcomes in patients with END (odds ratio 0.068, 95% confidence interval 0.006-0.774; P=0.030). Conclusions:The incidence of END is higher in patients with minor ischemic stroke caused by LVO, and internal carotid artery occlusion is an independent risk factor for the occurrence of END. Providing REVT after END may improve the clinical outcomes of patients with END at discharge.
8.Guideline for clinical perioperative care of orthopedic trauma patients in the new stage of novel corona virus infection (version 2023)
Chenchen YAN ; Bobin MI ; Wu ZHOU ; Faqi CAO ; Yun SUN ; Mengfei LIU ; Yiqiang HU ; Guandong DAI ; Dianying ZHANG ; Guodong LIU ; Zhiyong HOU ; Kun ZHANG ; Bin YU ; Jinmin ZHAO ; Xinlong MA ; Xieyuan JIANG ; Xinbao WU ; Jican SU ; Peifu TANG ; Baoguo JIANG ; Yingze ZHANG ; Guohui LIU
Chinese Journal of Trauma 2023;39(4):309-317
As the National Health Commission changes the management of novel corona virus infection, the situation and preventive policies for controlling the epidemic have also entered a new stage in China. Perioperative care strategies for orthopedic trauma such as designated isolation and nucleic acid test screening have also been adjusted in the new stage. Based on the perioperative work experiences in the new stage of epidemic from the frontline anti-epidemic staff of orthopedics in domestic hospitals and combined with the literature and relevant evidence-based medical data in perioperative care of orthopedic trauma patients under the current anti-epidemic policies at home and abroad, Chinese Orthopedic Association and Chinese Society of Traumatology organized relevant experts to formulate the Guideline for clinical perioperative care of orthopedic trauma patients in the new stage of novel corona virus infection ( version 2023). The guideline summarized 16 recommendations from the aspects of preoperative diagnosis and treatment, infection prevention, emergency operation and postoperative management to systematically standardize the perioperative clinical pathways, diagnosis and treatment processes of orthopedic trauma in the new stage of novel corona virus infection, so as to provide a guidance and reference for hospitals at all levels to carry out relevant work in current epidemic control policies.
9.Automatic radiolabeling of the norepinephrine transporter targeted tracer 18F-mFBG and evaluation of 18F-mFBG PET/CT imaging in pheochromocytoma
Weijian YE ; Jie MA ; Chenchen DONG ; Junqi HU ; Guocong LI ; Junjie WEI ; Ke ZHONG ; Hao XU ; Lu WANG
Chinese Journal of Nuclear Medicine and Molecular Imaging 2023;43(9):543-548
Objective:To fulfill the automatic radiolabeling of the norepinephrine transporter (NET) trancer 18F-meta-fluorobenzylguanidine (mFBG), and explore the 18F-mFBG PET/CT imaging effect of pheochromocytoma. Methods:On the basis of the chemical structure of mFBG, a spirocyclic iodonium ylide was used as the precursor to undergo a 3-step reaction sequence (radiofluorination, deprotection and neutralization) on AllinOne synthesis module. Purification by high performance liquid chromatography and formulation were conducted to generate 18F-mFBG. The corresponding quality control tests of 18F-mFBG product was performed. Afterwards, a postoperative patient with pheochromocytoma underwent 18F-mFBG PET/CT imaging. Results:The radiosynthesis was accomplished within 70 min, and 18F-mFBG was obtained in (17.8±2.4)% non-decay-corrected radiochemical yield ( n=5), with radiochemical purity >97% and molar activity >59.2 GBq/μmol. Sterility test, bacterial endotoxins test, abnormal toxicity test and the acetonitrile residue all met the requirements of Pharmacopoeia of the People′ s Republic of China (2020 Volume Ⅳ). The 18F-mFBG PET/CT imaging disclosed high uptake in pheochromocytoma and clear localization of lesions. Conclusions:The automatic radiolabeling of the NET targeted tracer 18F-mFBG is successfully realized by commercially available synthesis module, and the production quality meets all requirements for clinical translation. 18F-mFBG has a potential to image neuroendocrine lesions in clinical setting.
10.Correlation between iodine nutrition and thyroid function of pregnant women in Xinjiang Uygur Autonomous Region
Tayier RISHALAITI ; Yimu YUAN ; Pinjiang MA ; Qin LIN ; Shunhua WU ; Chenchen WANG
Chinese Journal of Endemiology 2023;42(2):134-138
Objective:Through the detection of iodine nutrition level and thyroid function of pregnant women in Xinjiang Uygur Autonomous Region (Xinjiang), to preliminary study the pregnant women's iodine nutrition level, thyroid function status and the relationship between the two and influencing factors.Methods:From March to June in 2020, stratified cluster random sampling method was adopted. Two counties (cities) in Southern and Northern Xinjiang were selected as survey sites, and about 100 pregnant women (a total of 412) were selected from each county (city) as survey subjects. Random urine samples and blood samples were collected to detect urinary iodine and serum thyroid function indicators [thyroid stimulating hormone (TSH), free triiodothyronine (FT 3), free thyroxine (FT 4), anti-thyroglobulin antibody (Tg-Ab) and anti-thyroid peroxidase antibody (TPO-Ab)]. Results:The median and interquartile range [ M ( Q1, Q3)] of pregnant women's urinary iodine was 228.4 (143.15, 327.95) μg/L. Serum FT 3, FT 4 and TSH levels [ M ( Q1, Q3)] were 4.22 (3.92, 4.61), 13.79 (12.63, 15.26) pmol/L and 1.82 (1.26, 2.52) mU/L, respectively. The overall positive rates of Tg-Ab and TPO-Ab were 5.61% (23/412) and 11.95% (49/412), respectively. The positive rates of Tg-Ab and TPO-Ab in Southern and Northern Xinjiang were 4.78% (10/209), 10.05% (21/209), 6.40% (13/203) and 13.79% (28/203), respectively. The positive rates of Tg-Ab and TPO-Ab in Northern Xinjiang were higher than those in Southern Xinjiang, but the difference was not statistically significant (χ 2 = 1.31, 2.17, P > 0.05). The positive rate of TPO-Ab in pregnant women was the influencing factor of abnormal thyroid function, and the odds ratio ( OR) [95% confidence interval ( CI)] was 3.22 (1.31 - 7.93). Conclusions:Pregnant women in Xinjiang are generally at an appropriate level of iodine, but the state of thyroid function still needs continuous attention. In addition, it is necessary to strengthen the thyroid function examination of pregnant women with positive thyroid autoantibodies to prevent and control the occurrence of abnormal thyroid function in pregnant women.

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