Objective:To investigate the expression of HBB in anaplastic thyroid cancer(ATC)cells and its regulatory effect on proliferation,invasion,migration and apoptosis of ATC cells and its potential mechanism.Methods:The expression of HBB in thyroid cancer and paracancerous tissues was analyzed through TIMER database.The correlation between the expression of HBB and the overall survival time of thyroid cancer patients was analyzed through KM-plotter database.The expression of HBB mRNA in ATC cells was detected by RT-qPCR.The HBB knockout or overexpression plasmid was transfected into ATC cells.The expression of HBB protein was detected by Western blot.The proliferation activity was detected by CCK-8 assay.The migration and invasion ability was detected by Transwell assay.The apoptosis was detected by flow cytometry.The expression of β-catenin was detected by Western blot.Results:The expression of HBB was low in thyroid cancer,and the overall survival time of patients with high expression of HBB was high.The expression of HBB protein was down-regulated in ATC cells.Knockout of HBB increased the ability of proliferation,migration and invasion of ATC cells and the expression of β-catenin protein,and inhibited apoptosis.However,overexpression of HBB decreased the ability of proliferation,migration and invasion of ATC cells and the expression of β-catenin protein,and promoted apoptosis.Conclusions:High HBB expression is associated with higher overall survival in patients with thyroid cancer.It may inhibit the proliferation,migration and invasion of ATC cells and promote apoptosis through Wnt/β-catenin signal pathway.

Objective To explore the effects of the long non-coding RNA(lncRNA)NK2 homeobox 1-antisense RNA 1(NK2-1-AS1),which mediates the microRNA(miR)-96-5p/PR domain-containing protein 16(PRDM16)axis,on anaplastic thyroid cancer(ATC)cell proliferation,migration,and invasion in vitro and transplanted tumor growth in vivo.Methods The differentially expressed lncRNA NKX2-1-AS1 in ATC tissues and cells,its target miRNA miR-96-5p,and its downstream target gene PRDM16 were screened using a bioinformatics analysis.The dual-luciferase reporter assay validated the relationship between NKX2-1-AS1 and miR-96-5p as well as the connection between miR-96-5p and PRDM16.Western blotting was performed to detect the effect of miR-96-5p overexpression on PRDM16 in CAL-62 cells overexpressed with NKX2-1-AS1.Plate clone formation,scratch,and Transwell assays were used to detect the effects of PRDM16knockdown on the proliferation,migration,and invasion of CAL-62 cells overexpressing NKX2-1-AS1.CAL-62 cells were injected subcutaneously into nude mice and the effect was observed of PRDM16knockdown on the growth of transplanted tumors of CAL-62 cells overexpressing NKX2-1-AS1.Results The bioinformatics analysis revealed that the NK2-1-AS1/miR-96-5p/PRDM16 axis was involved in regulating ATC development.The dual-luciferase reporter assay demonstrated that NKX2-1-AS1 bound to miR-96-5p and miR-96-5p bound to PRDM16.NKX2-1-AS1 overexpression upregulated PRDM16 protein expression in CAL-62 cells,while miR-96-5p overexpression reversed this phenomenon.NKX2-1-AS1 overexpression inhibited CAL-62 cellular proliferation,migration,and invasion in vitro and transplanted tumor growth in vivo,while knocking down PRDM16 reversed these phenomena.Conclusion NK2-1-AS1 may compete with miR-96-5p as an endogenous RNA to bind to its downstream target gene,PRDM16,and upregulate its expression,thus inhibiting ATC cell proliferation,migration,and invasion in vitro and transplanted tumor growth in vivo.

Objective To explore the effect of transcription factor E2F7 on the proliferation,migration,invasion,and tumor growth of anaplastic thyroid cancer(ATC)cells in vitro and to elucidate the underlying mechanisms.Methods Lentivirus transfection was used for a stable E2F7 knockdown in CAL-62 cells,and real-time PCR was used to detect E2F7 expression in these cells to verify the trans-fection efficiency.CAL-62 cells were divided into sh-NC and sh-E2F7 groups,and cell proliferation was measured using the CCK-8 assay,whereas cell migration and invasion abilities were measured using the Transwell assay.CAL-62 cells were subcutaneously injected into nude mice to observe tumor growth.The EPD website predicted an E2F7 binding site on the CXCL5 promoter,and the dual-lucif-erase reporter gene assay measured the effect of E2F7 knockdown on the luciferase activity of the CXCL5 promoter.The impact of E2F7 knockdown on CXCL5 levels in CAL-62 cells was assessed through real-time PCR and ELISA.Further,CAL-62 cells were divided into sh-E2F7+vector and sh-E2F7+CXCL5 groups to study the effects of CXCL5 overexpression on cell proliferation,migration,invasion,and the CXCR2/ERK signaling pathway following E2F7 knockdown.Results E2F7 knockdown inhibited CAL-62 cell proliferation,migra-tion,and invasion in vitro and tumor growth in vivo.The CXCL5 promoter has an E2F7 binding site,and E2F7 knockdown reduced the luciferase activity of the CXCL5 promoter.CXCL5 overexpression reversed the inhibitory effect of E2F7 knockdown on cell proliferation,migration,invasion,and the CXCR2/ERK signaling pathway in CAL-62 cells.Conclusion E2F7 promotes ATC cell proliferation,migra-tion,invasion,and tumor growth in vitro by activating the CXCL5/CXCR2/ERK signaling pathway mediated by CXCL5 transcription.

Objective: To observe the effects of tendon-regulating and stretching manipulation plus JIN's three-needle therapy for the shoulder on pain and shoulder joint function in subacromial impingement syndrome (SIS). Methods: Eighty patients with SIS were recruited and divided into a control group and a treatment group by the random number table method, with 40 cases in each group. The control group was given JIN's three-needle therapy for the shoulder, and the treatment group received additional tendon-regulating and stretching manipulation. The visual analog scale (VAS) score and constant-Murley score (CMS) were observed before and after the intervention, and the total effective rate was also observed. Results: The total effective rate was 92.5％ in the treatment group versus 70.0％ in the control group, and the difference was statistically significant (P<0.05). The VAS score and CMS changed notably after treatment in both groups (P<0.05), and the improvements were markedly greater in the treatment group than in the control group (P<0.05). Conclusion: Tendon-regulating and stretching manipulation plus JIN's three-needle therapy for the shoulder can facilitate the relief of pain and the improvement of shoulder joint function in SIS patients.

This report describs 2 domestic cases of tacrolimus poisoning in kidney transplant recipients due to overexposure of tacrolimus caused by nirmatrelvir/ritonavir for SARS-CoV-2 infection.Phenytoin sodium is prescribed for inducing CYP3A enzyme.It is intended for providing references for formulating and adjusting treatment protocols for tacrolimus overexposure and related toxicity in kidney transplant recipients caused by nirmatrelvir/ritonavir.

Objective:To summarize the patient profiles and therapeutic efficacies of ABO-incompatible living-related kidney transplantations at 19 domestic transplant centers and provide rationales for clinical application of ABOi-KT.Methods:Clinical cases of ABO-incompatible/compatible kidney transplantation (ABOi-KT/ABOc-KT) from December 2006 to December 2009 were collected. Then, statistical analyses were conducted from the aspects of tissue matching, perioperative managements, complications and survival rates of renal allograft or recipients.Results:Clinical data of 342 ABOi-KT and 779 ABOc-KT indicated that (1) no inter-group differences existed in age, body mass index (BMI), donor-recipient relationship or waiting time of pre-operative dialysis; (2) ABO blood type: blood type O recipients had the longest waiting list and transplantations from blood type A to blood type O accounted for the largest proportion; (3) HLA matching: no statistical significance existed in mismatch rate or positive rate of PRA I/II between two types of surgery; (4) CD20 should be properly used on the basis of different phrases; (5) hemorrhage was a common complication during an early postoperative period and microthrombosis appeared later; (6) no difference existed in postoperative incidence of complications or survival rate of renal allograft and recipients at 1/3/5/10 years between ABOi-KT and ABOc-KT. The acute rejection rate and serum creatinine levels of ABOi-KT recipients were comparable to those of ABOc-KT recipients within 1 year.Conclusions:ABOi-KT is both safe and effective so that it may be applied at all transplant centers as needed.

Objective To explore the clinical significance of tumor pathological length in evaluation of prognosis of esophageal squamous cell carcinoma.Methods 421 patients with esophageal squamous cell carcinoma were selected as study subjects.Each patient's data of gender,age,pathological examination(tumor pathological length),survival time,cancer stage,operation mode were collected.The patients were divided into two groups:≤4 cm group(189 cases) and >4 cm group(232 cases), and the constituent ratios of gender,age,or operation mode and the survival rate between the two groups were analyzed.Results There were no significant differences in gender composition,age structure and operation mode between the two groups(x2=2.03,2.41,0.06,all P>0.05);There was positive correlation between tumor pathological length and TNM stage(CMH=68.07,P<0.01).The 25%,50% and 75% quantiles of survival period in the group whose tumor length was less than or equal to 4 cm were 28,40 and 58 months,respectively,which in the group whose tumor length was more than 4 cm were 19,31 and 61 months,and there was significantly difference between the two groups(x2=44.88,P<0.01), the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm.In the further stratification analysis by TNM stage,the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm in theⅠstage patients(x2=5.61,P<0.05);the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm in theⅡstage patients(x2=4.62,P<0.05);the group whose tumor length was less than or equal to 4 cm had higher survival rate than the group whose tumor length was more than 4 cm in the Ⅲ stage patients(x2=4.88,P<0.05).Conclusion Tumor pathological length is an effective prognostic factor for esophageal squamous cell carcinoma,and the critical value can be determined as 4 cm.

Objective To explore the prognostic factors of portal hypertension treated with devascularization.Methods A total of 397 patients with portal hypertension underwent devascularization in Nanjing Drum Tower Hospital from February 1993 to April 2014,among which there were 242 male and155 female patients with median age of 48 years.The perioperative data were retrospectively collected.Logistic regression was used to find the risk factors which affect the operative complications.Follow-up evaluation was in progress regularly.Kaplan-Meier survival curve,Log-rank test and Cox regression model were used to find out factors which affect the long-term results.Results All together 397 patients underwent devascularization,in whom 8 patients died perioperative,389 patients discharged successfully.Logistic regression showed that age (≥ 48 years) (X2 =4.559,OR =2.048,P =0.033),red color sign before surgery (x2 =4.959,OR =2.129,P =0.026) and without portosystemic collateral vessels reserved (x2 =13.348,OR =5.122,P =0.000) were risk factors of perioperative complications.The follow-up time was (5.7±4.6) years.Totally 27 patients were lost from follow-up,103 patients died for the disease during follow-up.The survival rate at 1-,3-,5-,10-,15-and 20-years was 93.6％,86.9％,80.1％,59.3％,54.1％ and 38.5％ respectively.Univariate analysis showed that gender (male),age (≥ 48 years),hemorrhage before surgery (≥500 ml per time),hepatitis virus and without portosystemic collateral vessels reserved were risk factors of the long-term survival (P ＜ 0.05).Cox regression analysis showed that age (≥48 years) (X2 =9.850,RR =1.904,P =0.002),hemorrhage before surgery (≥ 500 ml per time)(X2 =34.402,RR =3.273,P =0.000),hepatitis virus (X2 =7.573,RR =2.525,P =0.006) and without portosystemic collateral vessels reserved (x2 =5.905,RR =１.889,P =0.015) were independent risk factors that affect the long-term survival.Conclusion Devascularization with portosystemic collateral vessels reserved has favorable perioperative and long-term outcome,and it definitely is a very safe and effective technique for portal hypertension.

Objective To explore the prognostic factors of portal hypertension treated with devascularization.Methods A total of 397 patients with portal hypertension underwent devascularization in Nanjing Drum Tower Hospital from February 1993 to April 2014,among which there were 242 male and155 female patients with median age of 48 years.The perioperative data were retrospectively collected.Logistic regression was used to find the risk factors which affect the operative complications.Follow-up evaluation was in progress regularly.Kaplan-Meier survival curve,Log-rank test and Cox regression model were used to find out factors which affect the long-term results.Results All together 397 patients underwent devascularization,in whom 8 patients died perioperative,389 patients discharged successfully.Logistic regression showed that age (≥ 48 years) (X2 =4.559,OR =2.048,P =0.033),red color sign before surgery (x2 =4.959,OR =2.129,P =0.026) and without portosystemic collateral vessels reserved (x2 =13.348,OR =5.122,P =0.000) were risk factors of perioperative complications.The follow-up time was (5.7±4.6) years.Totally 27 patients were lost from follow-up,103 patients died for the disease during follow-up.The survival rate at 1-,3-,5-,10-,15-and 20-years was 93.6％,86.9％,80.1％,59.3％,54.1％ and 38.5％ respectively.Univariate analysis showed that gender (male),age (≥ 48 years),hemorrhage before surgery (≥500 ml per time),hepatitis virus and without portosystemic collateral vessels reserved were risk factors of the long-term survival (P ＜ 0.05).Cox regression analysis showed that age (≥48 years) (X2 =9.850,RR =1.904,P =0.002),hemorrhage before surgery (≥ 500 ml per time)(X2 =34.402,RR =3.273,P =0.000),hepatitis virus (X2 =7.573,RR =2.525,P =0.006) and without portosystemic collateral vessels reserved (x2 =5.905,RR =１.889,P =0.015) were independent risk factors that affect the long-term survival.Conclusion Devascularization with portosystemic collateral vessels reserved has favorable perioperative and long-term outcome,and it definitely is a very safe and effective technique for portal hypertension.

Objective To explore the differences between ABI 7500 and TL 988 fluorescent quantitative instrument for polymerase chain reaction. Methods Based on EP9-A2 Files, quantitative influenza A virus positive controls of five kinds of concentrations were detected by two kinds of fluorescence quantitative PCR instruments at the same time, and each sample was detected 20 times. The precision and sensitivity between the two kinds of instruments were compared. Results Inner group CV of TL 988 ranged from 0.68% to 1.52%. The CV for ABI 7500 ranged from 1.93% to 2.69%. The high (107, 106, 105 copies/mL) levels of IFVA positive control were detected by ABI 7500 and TL 988 instruments, and there were no significant difference between the results(P>0.05) while the low(104, 103 copies /mL)levels were de-tected by ABI 7500 and TL 988 instruments, and there was a significant difference between the results(P<0.05). When samples were detected at the level of 103 copies/mL, the positive detection rate was 100% with TL 988 instrument, but the positive detection rate was 0 with ABI 7500 instrument. The related index of ABI 7500 and TL 988 was better (r=0.998). Conclusion The performances in precision and sensitivity of TL 988 PCR instrument are better than ABI 7500 PCR.