Objective To investigate the effect of long non-coding ribonucleic acid cancer susceptibility can-didate gene 19(lncRNA CASC19)regulating the microRNA(miR)-490-3p/high mobility group protein A2(HMGA2)axis on the proliferation,migration,and chemotherapy resistance of colorectal cancer(CRC)cells.Methods Human normal colonic epithelial cells(FHC)and CRC cell lines(LOVO,HCT116,SW480)were cultured.LOVO cells were randomly divided into Control group,sh-NC group,sh-CASC19 group,sh-CASC19+anti-NC group and sh-CASC19+anti-miR-490-3p group.The lncRNA CASC19,miR-490-3p and HMGA2 mRNA expression were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).The relationship between lncRNA CASC19 and miR-490-3p and between miR-490-3p and HM-GA2 were detected by Dual luciferase assay.The cell proliferation ability was detected by cell counting kit-8(CCK-8)assay and colony formation assay.The cell migration ability was detected by cell scratch assay.The cell invasion ability was detected by Transwell assay.The cell metastasis-related proteins(MMP2,MMP9)and HMGA2 protein expression were detected by Western blot assay.The chemotherapy resistance was detec-ted by cisplatin and fluorouracil.Results The lncRNA CASC19 and HMGA2 mRNA expression increased in CRC cell lines,and miR-490-3p expression decreased.The lncRNA CASC19,miR-490-3p and HMGA2 mRNA expression in LOVO cells were the most significant,LOVO cells were selected for subsequent experiments.The dual luciferase assay showed that,after the transfection of lncRNA CASC19 and HMGA2,compared with mimic-NC group,the luciferase activity in miR-490-3p mimic group decreased(P＜0.05).Compared with sh-NC and Control groups,the survival rate,clone number,migration rate,invasion rate,MMP2,MMP9,HMGA2 mRNA and protein expression of LOVO cells in sh-CASC19 group were decreased,and the miR-490-3p ex-pression was increased(P＜0.05).Compared with sh-CASC19 group and sh-CASC19+anti-NC group,the survival rate,clone number,migration rate,invasion rate,MMP2,MMP9,HMGA2 mRNA and protein expres-sion of LOVO cells in sh-CASC19+anti-miR-490-3p group were increased,and the miR-490-3p expression was decreased(P＜0.05).The chemotherapy resistance experiment showed that,compared with Control group,the chemotherapy resistance sensitivity of LOVO cells in sh-CASC19 group increased(P＜0.05).Compared with sh-CASC19 group,the chemoresistance sensitivity of LOVO cells in sh-CASC19+anti-miR-490-3p group was decreased(P＜0.05).In the same group,with the increase of the concentration of fluorou-racil and cisplatin,the cell survival rate gradually decreased(P＜0.05).Conclusion Knockdown of lncRNA CASC19 can regulate miR-490-3 p/HMGA2 signaling pathway,inhibit the proliferation and migration of CRC cells,and increase the sensitivity of chemotherapy resistance.

OBJECTIVE: To explore the clinical and genetic characteristics of eight children with Primary hypertrophic cardiomyopathy (HCM). METHODS: Eight children with HCM admitted to the Department of Cardiology of Henan Children's Hospital from January 2018 to December 2021 were selected as the study subjects. Clinical data of the children were collected. Whole exome sequencing was carried out on two children, and trio whole exome sequencing was carried out on the remainder 6 children. Sanger sequencing was used to verify the candidate variants in the children and their parents, and the pathogenicity of the variants was evaluated based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). RESULTS: The patients had included 5 males and 3 females, with their ages ranging from 5 to 13 years old. The average age of diagnosis was (7.87 ± 4.8) years old, and the cardiac phenotype showed non-obstructive HCM in all of the patients. WES has identified variants of the MYH7 gene in 4 children, including c.2155C>T (p.Arg719Trp), c.1208G>A (p.Arg403Gln), c.1358G>A (p.Arg453His), and c.1498G>A (p.Glu500Lys). Based on the guidelines from the ACMG, the first 3 variants were classified as pathogenic, while c.1498G>A (p.Glu500Lys) was classified as likely pathogenic (PM1+PM2_Supporting+PM6+PP3), which was also unreported previously. The remaining four children had all harbored maternal variants, including MYL2: c.173G>A (p.Arg58Gln; classified as pathogenic), TPM1: c.574G>A (p.Glu192Lys) and ACTC1: c.301G>A (p.Glu101Lys)(both were classified as likely pathogenic), and MYBPC3: c.146T>G (p.Ile49Ser; classified as variant of uncertain significance). Seven children were treated with 0.5 ~ 3 mg/(kg·d) propranolol, and their symptoms had improved significantly. They were followed up until September 30, 2022 without further cardiac event. CONCLUSION Genetic testing can clarify the molecular basis for unexplained cardiomyopathy and provide a basis for clinical diagnosis and genetic counseling. Discovery of the c.1498G>A (p.Glu500Lys) variant has also expanded the spectrum of MYH7 gene mutations underlying HCM.
Female ; Male ; Humans ; Child ; Child, Preschool ; Adolescent ; Cytoskeletal Proteins ; Family ; Genetic Counseling ; Genetic Testing ; Cardiomyopathy, Hypertrophic/genetics*

To improve collagen production by recombinant Pichia pastoris, we applied Placket-Burman and Box-Behnken design to optimize the fermentation medium. Through Placket-Burman design, three variables in the medium (concentration of yeast extract, peptone and glycerol) were selected for having significant effect on cell dry weight. Through Box-Behnken design regression coefficients analysis, a secondary degree polynomial equation was established, and the optimum levels of the three variables were yeast extract 1.13%, peptone 1.61% and glycerol 0.86%. During the growth period, an average cell dry weight of 4.41 g/L was obtained after 12 h fermentation, increased by 26%. Through high density fermentation, the production of recombinant human collagen (Ⅲ) was up to 4.71 g/L in 22 L fermentor. The recombinant human collagen (Ⅲ) exhibited good results to repair acetic acid induced gastric ulcer in rats.

Objective The goal of this study is to compare the prognosis of recombinant tissue plasminogen activator (rt-PA) thrombolysis for middle cerebral artery (MCA) occlusion with patients with good and poor cerebral collateral circulation.Methods This retrospective study included 49 patients diagnosed with acute MCA occlusion and treated with rt-PA in the First Affiliated Hospital of Nanjing Medical University between October 1,2014 and February 1,2016.Patients were divided into good collaterals group (n =31) and poor collaterals group (n =18) according to their distribution of leptomeningeal arteries with CTA.Thirty day mortality rate,the incidence of symptomatic intracranial hemorrhage,24h and 30 day Stroke scores with National Institute of Health Stroke Scale (NIHSS) were compared between the two groups.Corrected chi-squared test,Fisher's exact test,or t test was used to statistical analysis as appropriate.Results The 30 day mortality rate of good collaterals group was significantly lower than that of poor collaterals group (0％ vs.16.7％,P ＜ 0.05).There were no significant differences in the incidence of symptomatic intracranial hemorrhage and 24h NIHSS score between the two groups (P ＞ 0.05),however,30 day NIHSS score of good collaterals group was significantly lower than that of poor collaterals group (7.2 ± 3.1 vs.9.6 ± 2.7,P ＜ O.05).Conclusion For patients with MCA occlusion and receiving intravenous thrombolysis,good cerebral collateral circulation may reduce their mortality and improve their clinical outcome after thrombolysis.However,good cerebral collateral circulation does not reduce the risk of symptomatic intracranial hemorrhage in those patients.

AIM:To study the expression of signal transduction molecules in the striatum G protein protein 4 (RGS4) and dopamine D2 receptor (D2) in conditioned place preference (CPP) rats treated with methamphetamine (meth).METHODS:METH dependence CPP model was established (1 week and 2 weeks of METH dependence groups),The protein expression of RGS4 and D2,inhibitory G protein alpha-subunit (Gαi),mitogenactivated protein kinase (MAPK) in striatum were determined by Western blotting (WB).The changes of cyclic adenosine monophosphate (cAMP) content in striatum of rats were determined by enzyme linked immunosorbent assay (ELISA).RESULTS:Compared with saline control group,the average time of rats in the methamphetamine-paired chamber for two groups was increased (P ＜ 0.05).Compared with saline control group,RGS4 protein expressions in the two METH dependent groups were reduced (P ＜0.01);compared with 1 week of METH dependence group,that of 2 weeks group was reduced significantly(P ＜ 0.05).D2,Gαi,MAPK protein and cAMP expressions in the two METH dependent groups were increased (P ＜ 0.01);compared with 1 week of METH dependence group,those of 2 weeks were increased significantly (P ＜ 0.05).CONCLUSION:RGS4 and D2 receptor signaling pathways in striatum have changed in METH dependent rats,RGS4 may be involved in the regulation of METH-dependent D2 receptor signaling pathway in METH dependent rats.

AIM:To observe the effects and mechanisms of hydroxyethylstarch (HES) 130/0.4 on no-reflow phenomenon after myocardial ischemia-reperfusion in rats.METHODS: SD rats were randomly divided into 4 groups:sham operation group , ischemia-reperfusion ( IR, treated with normal saline ) group, normal saline ischemia-reperfusion (NS-IR, treated with NS) group and HES ischemia-reperfusion (HES-IR, treated with HES) group.Myocardial infarct size and no-reflow range were determined by staining methods , and the activities of myocardial enzymes ( CK-MB, cTnI and MPO) were measured .Meanwhile , cardiac microvascular endothelial cells of the rat were cultured and divided into 4 groups:control group, hypoxia/reoxygenation (H/R) group, NS-H/R group and HES-H/R group.Acute ischemia reper-fusion models were simulated , and the concentration of calcium ions was measured .The relative cell activity was evaluated by CCK-8 assay, and the apoptotic rate was detected by flow cytometry .RESULTS:In HES-IR group, the myocardial in-farct size, the no-reflow zone, CK-MB, cTnI and MPO activity were all significantly lower than those in IR group ( P<0.05).In microvascular endothelial cells , the concentration of calcium ions and the apoptotic rate in HES-H/R group were significantly decreased, while the relative cell activity increased compared with H/R group (P<0.05).CONCLUSION:HES reduces no-reflow in acute myocardial ischemia-reperfusion .The mechanism may be involved in the inhibition of both the infiltration of neutrophils and the calcium overload of endothelial cells .

Objective To investigate the prevalence of hyperuricemia in health check-up population of Beijing suburb.Methods Total 1 336 rural residents in Nankou Township of Beijing received health check-up from July to Aug 2014,including 686 subjects aged 20-59 years (young/middle-aged group) and 650 subjects aged 60-96 years (elderly group).The blood pressure and body mass index (BMI) were measured;serum uric acid (SUA),fasting blood glucose (FBG) and blood lipids (TG,TC,HDL-C,LDL-C) were determined.The SUA levels ＞ 420 μmol/L for male and ＞ 360 μmol/L for female were defined as hyperuricemia.Results The four quartiles of SUA levels were 27.00-254.59 μmol/L (Q1),254.60-302.35 μmol/L (Q2),302.36-359.78 μmol/L(Q3) and 359.79-702.0 μmol/L (Q4).The prevalence of hyperuricemia was significantly higher in young/middle-aged group than that in elderly group [20.41％ (140/686) vs.13.85％ (90/650),x2 =10.08,P =0.001 5],the systolic blood pressure [SBP,(126.8±15.7) vs.(116.7±12.0)mmHg(1 mmHg=0.133 kPa),t=2.76,P=0.008],FBG [(7.40±4.10) vs.(6.11 ±2.03)mmol/L,t=2.12,P=0.036],TC [(5.52±1.10) vs.(5.23±1.00)mmol/L,t =2.04,P =0.045],LDL-C [(3.5 ±0.7) vs.(2.4 ±0.9)mmol/L,t =2.21,P =0.029]in young/middle-aged group were significantly higher than those in elderly group.BMI,FBG were significantly higher in Q4 than those in other quartiles [BMI:(26.44 ± 3.88) vs.(24.19 ± 3.37),(25.49±3.42) and (25.61 ±3.49)kg/m2,t =2.78,P=0.008;FBG:(8.19 ±1.52) vs.(6.34±1.34),(6.09 ± 1.51) and (6.40 ± 1.98) mmol/L,t =2.80,P =0.007].The triglyceride (TG) levels in group Q3 and Q4 [(1.85 ± 0.90) and (1.92 ± 0.44) mmol/L] were higher than those in Q1 and Q2 [(1.37 ±0.76) and (1.70 ±0.84) mmol/L,t =2.1,P =0.035].Only 9.57％ subjects (22/230)with hyperuricemia was not combined with metabolic disorder;subjects combined with one and two metabolic disorders accounted for 20.87％ (48/230) and 69.57％ (160/230),respectively.Conclusion Screening for hyperuricemia is important for comprehensiye treatment and management of hyperuricemia in rural residents,especially in the young and middle-aged population.

OBJECTIVETo study the extensibility and retractility of the surgical margins in digestive system neoplasms.

METHODSThe length difference of the digestive tract was measured in vivo and in vitro under different conditions. Five cm of the stomach, small intestine and large bowel and 2 cm of the esophagus were measured as standard control length in vivo just before resection. The length was measured with a ruler under pull of 500 g and 1 000 g in vivo, in fresh status in vitro, and 10% formaldehyde fixed for 6 - 8 h, 12 - 24 h and 48 - 72 h. The extension or retraction ratio was calculated. The length difference was divided by the natural length in vivo.

RESULTSSeventeen cases of the esophagus, 18 cases of the stomach, 15 cases of the small intestine and 25 cases of the large bowel were measured under pull of 500 g and 1 000 g. The esophagus extended 16.5% and 30.5%, stomach 15.0% and 22.6%, small intestine 66.4% and 120.0%, large bowel 36.0% and 56.0% respectively. In natural status ex vivo, the esophagus retracted 44.5%, stomach 13.6%, small intestine 11.4% and large bowel 15.6% respectively; they continue to retract after 10% formaldehyde fixation until 12 - 24 h later. If the length of surgical margin of the fresh specimen ex vivo was x, the natural length of margin in vivo of the esophagus would be 1.80 x, stomach 1.16 x, small intestine 1.13 x, and large bowel 1.18 x. If formaldehyde fixation for 6 - 8 h, the natural length of surgical margin in vivo of the esophagus would be 1.82 x, stomach 1.41 x, small intestine 1.22 x, large bowel 1.55 x. If formaldehyde fixation for 12 - 24 h, the surgical margin length in vivo of the esophagus would be 2.22 x, stomach 1.43 x, small intestine 1.28 x, and large bowel 1.57 x.

CONCLUSIONThe length of surgical margin of digestive system cancers varied under different conditions, and the evaluation of surgical margin during surgery should be performed under natural status in vivo.

Adult ; Aged ; Digestive System Neoplasms ; surgery ; Digestive System Surgical Procedures ; Female ; Humans ; Male ; Middle Aged