N6-Methyladenosine (m6A) modification is a crucial post-transcriptional regulatory mechanism and the most abundant and highly conserved RNA epigenetic modification in eukaryotes. Previous studies have indicated the involvement of m6A modification in various tissue regeneration processes, including liver regeneration. Vir-like m6A methyltransferase associated protein (VIRMA) is an m6A methyltransferase with robust methylation capability. However, its role in liver regeneration remains poorly understood. In this study, we generated liver-specific Virma knockout mice using the Cre-loxP system and investigated the biological functions of VIRMA in liver regeneration using both the Associating Liver Partition and Portal vein Ligation for Staged Hepatectomy (ALPPS) mouse model and the carbon tetrachloride (CCl₄) mouse model. The expression level of VIRMA was rapidly up-regulated after ALPPS surgery and gradually down-regulated during liver repair. Virma deficiency significantly impaired liver regeneration capacity and disrupted cell cycle progression. Methylated RNA immunoprecipitation sequencing (MeRIP-seq) analysis revealed that Shq1 is an effective downstream target of VIRMA-mediated m6A modification. The upregulation of Shq1 enhanced the proliferation ability of cells, which was attenuated by the specific AKT inhibitor ipatasertib. Supplementation of Shq1 in vivo alleviated the liver cell proliferation inhibition caused by Virma deficiency. Furthermore, the m6A-binding protein heterogeneous nuclear ribonucleoprotein a2b1 (HNRNPA2B1) enhanced the mRNA stability of Shq1. Mechanistically, Virma deficiency resulted in decreased m6A modification on Shq1 mRNA, leading to reduced binding ability of m6A-binding protein HNRNPA2B1 with Shq1, thereby decreasing the mRNA stability of Shq1 and reducing its protein expression level. Downregulation of Shq1 inhibited the PI3K/AKT pathway, thereby suppressing cell proliferation and cell cycle progression, ultimately impeding liver regeneration. In summary, our results demonstrate that VIRMA plays a critical role in promoting liver regeneration by regulating m6A modification, providing valuable insights into the epigenetic regulation during liver regeneration.

Background and purpose:Chimeric antigen receptor T(CAR-T)cell therapy has shown remarkable efficacy in treating hematological and lymphatic system tumors,but its effectiveness in solid tumors is relatively poor,which is partly attributed to target selection.For Ewing sarcoma(ES),CD99 can be a potential target for CAR-T cells.However,due to T cells'endogenous expression of CD99 protein,CAR-T cells targeting CD99 face limitations in their expansion capacity in vitro.This study aimed to identify the optimal conditions for preparing CD99 CAR-T cells by incorporating CD99 knockdown short hairpin RNA(shRNA),optimizing the multiplicity of infection(MOI)for lentiviral transduction,and screening for the best culture medium and container for CAR-T cell expansion.Methods:shRNA sequences were screened to enhance the expansion capacity of CD99 CAR-T cells.Different MOI,culture media,and containers were used to assess CAR-T cell transduction efficiency,cell viability,proliferation capacity,specific killing ability,and interferon-γ(IFN-γ)release levels under various conditions,in order to identify the optimal cell preparation conditions.Results:The expansion level of KO-CD99 CAR-T cells obtained through shRNA knockdown was significantly higher than that of CD99 CAR-T cells[(16.40±0.40)vs(6.33±1.53),P<0.01].The optimal expansion effect was observed when the transduction MOI was between 0.25 and 1.0,and OptiVitro was used as the culture medium.CAR-T cells cultured in ventilated flasks exhibited significantly higher expansion rates compared to cells cultured in bags[MOI=0.25:(50.23±3.32)vs(13.02±4.82);MOI=0.50:(49.96±0.83)vs(18.25±2.88);MOI=1.00:(48.27±5.08)vs(13.16±6.26);P<0.01],with better cell phenotype and higher specific killing ability.Conclusion:KO-CD99 CAR-T cells obtained through shRNA technology can achieve stable expansion.Based on the optimization of expansion conditions,KO-CD99 CAR-T cells exhibit superior expansion capacity and a higher proportion of memory T cells when the MOI is between 0.25 and 1.00,OptiVitro is used as the culture medium,and ventilated flasks are used as the culture container.These findings lay a solid foundation for further clinical trials of CD99 CAR-T cell therapy for ES.

Objective:To investigate the prevention and risk factors of deep vein thrombosis (DVT) in the lower extremity after medial open wedge high tibial osteotomy (HTO).Methods:A total of 128 patients who underwent medial open wedge HTO in the Third Hospital of Hebei Medical University from January 2020 to October 2022 were retrospectively analyzed, including 45 males and 83 females, aged 59.3±6.8 years (range, 44-87 years). Postoperative anticoagulation with enoxaparin sodium was applied at a randomized dose of 4,000 AXaIU/d or 6,000 AXaIU/d. Gender, age, history of chronic diseases (hypertension, diabetes), smoking history, body mass index, and body fat percentage were collected. On admission, the risk of DVT was assessed using the Caprini scale and calf circumference was measured. Hemoglobin, D-dimer, antithrombin III, activated partial thromboplastin time (APTT), prothrombin time (PT), fibrinogen (FIB), fibrinogen degradation products (FDP), glutathione, glutathione, urea, creatinine, uric acid were recorded. Patients were divided into DVT group and non-DVT group according to whether DVT occurred after operation. Binary logistic regression was used to analyze the independent risk factors of DVT after HTO. The receiver operating characteristic (ROC) curve was plotted, and the area under curve (AUC) was calculated to analyze the predictive value of the postoperative Caprini scale in the occurrence of DVT after HTO.Results:A total of 128 patients were enrolled, 83 patients were treated with enoxaparin sodium 4 000 AXaIU/d and 45 patients were treated with enoxaparin sodium 6 000 AXaIU/d. According to the results of color Doppler examination of bilateral lower extremity veins on the third day after operation, DVT occurred in 39% (50/128) of patients, including 39 cases of calf intermuscular thrombosis, 6 cases of peroneal vein thrombosis, 4 cases of posterior tibial vein thrombosis, and 1 case of popliteal vein thrombosis. DVT occurred in 36% (30/83) of patients receiving 4 000 AXaIU/d enoxaparin sodium and 44% (20/45) of patients receiving 6 000 AXaIU/d enoxaparin sodium, with no statistically significant difference (χ 2=0.84, P=0.358). Univariate analysis showed that smoking history, postoperative Caprini scale≥8, and female may be associated with the development of DVT after HTO ( P<0.05). They were included in the binary logistic regression, and the results showed that postoperative Caprini scale≥8 was an independent risk factor for DVT after HTO. The ROC curve of postoperative Caprini scale for predicting DVT after HTO was drawn, and the AUC was 0.847 (95% CI: 0.73, 0.96), the optimal cut-off value was 8, and the sensitivity and specificity were 84.2% and 77.6%, respectively. Conclusion:Caprini scale≥8 is an independent risk factor for DVT after medial open wedge HTO. Caprini scale has a good value in predicting the occurrence of DVT after HTO. The recommended dose of enoxaparin sodium is 4 000 AXaIU/d for the prevention of postoperative DVT, and increasing the dose is not associated with a decreased risk of DVT.

[ ABSTRACT] AIM:To investigate the effect of HOX transcript antisense RNA ( HOTAIR) on the migration and invasion abilities of liver carcinoma HepG 2 cells.METHODS:The expression of phosphoinositide-3-kinase regulatory sub-unit 3 (PIK3R3) in the liver cancer and normal liver tissues was detected by immunohistochemistry .The efficiency of gene silencing of HOTAIR or PIK3R3 by LV3-shHOTAIR or LV3-shPIK3R3 was determined by qPCR and Western blot .The mi-gration and invasion abilities of HepG 2 cells after silencing of HOTAIR and PIK3R3 were measured by wound healing assay and Transwell Matrigel invasion assay .The expression of miR-214 after silencing of HOTAIR and PIK3R3 was analyzed by qPCR.The expression of HOTAIR and PIK3R3 in the HepG2 cells was also evaluated by qPCR after transfected with miR-214 mimics or miR-214 inhibitor .Dual-luciferase reporter assay system was used to determine the regulatory effect of miR-214 on HOTAIR and PIK3R3 expression.RESULTS:PIK3R3 expression increased significantly in the liver cancer tissues compared with normal liver tissues .The abilities of invasion and metastasis of hepatocellular carcinoma were reduced after silencing of HOTAIR and PIK3R3.miR-214 expression was increased when silencing of HOTAIR and PIK3R3 was per-formed.HOTAIR and PIK3R3 expression was reduced after transfection with miR-214 mimics.HOTAIR and PIK3R3 ex-pression was increased after transfection with miR-214 inhibitor.The results of dual-luciferase reporter assay test showed that miR-214 directly regulated HOTAIR and PIK3R3 transcription activity .CONCLUSION: HOTAIR regulates the ex-pression of PIK3R3 through miR-214, thus promoting the migration and invasion abilities in the liver cancer cells .

Objective To study the effects of Trichinella spiralis (T.spiralis) infection on mice with oxazolone (OXZ)-induced colitis and the possible immunologic mechanism.Methods Female BALB/c mice at age 6-8 weeks were randomly divided into four groups (A to D).Each mouse in groups B and D was infected with T.spiralis strains.Twenty-one days after T.spiralis infection,the mice in groups A and B were treated with 50％ ethanol solution,while those in groups C and D were treated with OXZ to induce the murine model of colitis.Several parameters including survival rate,disease activity index (DAI),macroscopic damage and histological score,myeloperoxidase (MPO) activity and the expression of cytokines (IFN-γ,IL-4 and IL-10) in colonic and splenic tissues at mRNA and protein levels were measured 3 and 7 days after modeling.Results No significant differences in the survival rate,DAI score,macroscopic damage score,histological score,MPO activity were observed between mice from groups C and D (P＞0.05).Pre-exposing the mice to T.spiralis strains neither alleviated the mucosal damages nor aggravated the condition of colitis.Compared with group C,the expression of IFN-γ on the third day and the expression of IFN-γ,IL-4 and IL-10 on the seventh day at mRNA and protein levels in colon and spleen tissues were significantly increased in mice treated with T.spiralis and OXZ (P＜0.05).The expression of IL-10 at transcriptional level in spleen tissues on the third day was higher than that of group C (P＜0.05).The expression of IL-4 and IL-10 at protein level in colon tissues on the third day were significantly up-regulated as compared with those of group C (P＜0.05).Conclusion The severity of OXZ-induced colitis in the murine model was neither alleviated nor aggravated by pre-exposing the mice to T.spiralis strains.High doses of IL-10 not only weakened the regulatory effects on Th2 responses,but also induced the production of proinflammatory cytokines,resulting in a new drift of Th1/Th2 without aggravating Th2 responses.Further investigation on the mechanism of T.spiralis-induced over-expression of IL-10 should be conducted,which might increase the practicability of using T.spiralis strains against OXZ-induced colitis.

Objective To research the clinical outcome of atypical glandular cell (AGC)according to various subtypes of HPV infection and histological pathology results.Methods The data of the liquid-bases cytology (LBP),HPV infection and histology in 102 cases of AGC at the gynecology outpatient department of our hospital from January 1 ,2009 to February 28,2014 were collected and performed the analysis on their clinical outcomes.Results Among 67 218 cases of LBP detection,102 cases were AGC with the total incidence rate of 0.15%.In the cases of AGC-NOS,67 cases were normal or benign lesions,11 cases were precancerous lesion and malignant lesions;in the cases of AGC treading to tumor,the benign,precancerous and malignant lesions were in 7,14 and 3 ca-ses respectively.At the same time in the cases of AGC-NOS,HPV infection was in 64 cases,in which 57 cases were high risk infec-tion(type 16,52,45)and 7 cases were low risk infection(type 6,11 ).The single infection,double infection and multiple infection were in 54,6 cases respectively;in the cases of AGC trends to neoplasm,HPV infection was in 19 cases,in which 18 cases were high risk infection(type 52,16,18)and 1 case was low risk infection(type 6),single infection and double infection were in 15 cases and 4 cases respectively.Conclusion AGC may play an important role for the forecast of cervical malignant lesions.The results of differ-ent HPV subtypes infection in AGC related tumors also play a certain role in the prediction of cervical neoplasia.Their combined a-nalysis is the important signal for judging the occurrence of gynecological cervical precancerous lesion and malignant tumor,i.e., AGC combining with the corresponding HPV subtype infection not only can make a judgement for the cervical lesions,but also pro-vides the basis for predicting the high risk existence of gynecological malignant tumor and provides constructive suggestions for Chi-na regional cervical carcinoma vaccine manufacturing and promotion.

Objective To comparatively analyse the effects and methods for the treatment of the medial collateral ligament injury ( Ⅲ degree injury) of knee by direct suture repair and autologous semitendinosus transferred to reconstruct the repair.Methods Sixty-two cases of medial collateral ligament injury were treated by direct suture repair (suturing group,23 cases) and autologous semitendinosus transferred to reconstruct the repair (rebuild group,39 cases) with knee joint injury from March 2002 to September 2009.All cases of knee joints functional assessment were evaluated according to the Lysholm-Scale upgrades function scoring.Compared the effectiveness for stability measurement of genu valgum stress test 6 months after operation.Results All patients were followed up for 6 - 60 ( 12.4 ± 4.8) months.The outcome according to the Lysholm-Scale upgrades function scoring,the excellent and good rate was 65.2%( 15/23 ) with suturing group and 92.3% (36/39) with rebuild group.The curative effectiveness of stabilization for knee joint's medial collateral,as following:the excellent and good rate was 65.2% (15/23)with suturing group and 97.4%(38/39) with rebuild group.The curative effectiveness of stabilization for knee joint's posteromedial,as following the excellent and good rate was 60.9% (14/23) with suturing group and 92.3% (36/39) with rebuild group.There were statistical significance in excellent and good rate between the two groups (P ＜ 0.05 ).Conclusions The treatment of reconstruction medial collateral ligament injury with autologous semitendinosus transfer could provide enough tension to direct suture joint capsule,more robust on anatomy.It can play to inhere biological effect.The therapy of anatomy reconstruction is very satisfactory,has merits of firm fixation,and the method is simple,convenient and available,which is an ideal one.

Objective To retrospectively evaluate the incidence and treatment of fungous infection compli-cations after hematopoietic stem cell transplantation. Methods The incidence, pathogenic microorganism, prophy-laxis,treatments of infectious complications in 150 patients, who accepted hematopoietic stem cell transplantation from September 1990 to Martch 2000 in our hospital were analyzed. Results The incidence of infectious complica-tions was 89.3% (134/150) in all 150 cases. Three patients (2%) died of the fungal infection. The incidence of the fungal infections was 32.5% (26/80) in patients who accepted treatment with impenem or/and ceftazidine, and 15.7% (11/70) in other patients without the above treatment (P<0.05). 12 fungal infection cases were treated with small-dosage of amphotericin B(10 mg/d) ,with the healing rate was 100%. Conclusion The strong antibac-terial prophylaxis can't reduce the incidence of infection ,but may increase the risk of fungal infection;small-dosage of amphotericin B is a new effective way to treat fungal infection.

In this paper is reported a new approach for the treatment of sarcoma--electroporation therapy. Electroporation can accelerate pharmacal molecules into cytoplasm by transient electromagnetic pulses. We have utilized the phenomenon of electroporation treating the S-180 sarcomas in the hind legs of the Kunming mice by intratumoral injection of anti-tumor agent at low dose. From the experiment, we learned that this approach can bring about remarkable effect. The technical procedure is easy to do and easy to control. Especially, it is useful in curing the flat tumor and has little untoward side effect. It deserves to be recommended as a new approach to treating the tumor in clinics.
Animals ; Antineoplastic Agents ; therapeutic use ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cyclophosphamide ; therapeutic use ; Electrochemotherapy ; methods ; Mice ; Neoplasm Transplantation ; Sarcoma 180 ; drug therapy ; pathology

This article reports the experiment studies on treating the S-180 sarcomas of KM mice with high-intensity electric pulse and antitumor drug (cyclophosphamide). The results showed that the experimental group of electric field combined with drug has the best effect on tumor, compared with the control group. In addition, electric field can inhibit the formation of vas Capillaries and decrease the supply of nutrition for tumor cell. In conclusion, electric field has inhibited the growth of tumor.
Animals ; Antineoplastic Agents ; therapeutic use ; Combined Modality Therapy ; Cyclophosphamide ; administration & dosage ; Electric Stimulation Therapy ; methods ; Injections, Intralesional ; Mice ; Sarcoma 180 ; pathology ; therapy ; Treatment Outcome