AIM:To investigate the effect of plate-let(PLT)on angiogenesis in hepatocellular carcino-ma and the intervention effect of Cinobufagin(CBG).METHODS:Firstly,we screened the suitable co-incubation ratio of PLT and hepatocellular carci-noma cells,prepared conditioned medium,and de-termined the half inhibitory concentration of Cinob-ufagin;then,we set up a control group(human um-bilical vein endothelial cells(EC)+conventional me-dium),a crosstalk group(EC+CM_HP(strip culture prepared by crosstalk of HUH7 and PLT)),and an in-tervention group(EC+CM_HP+CBG).The migration,tube-formation and sprouting capacity of EC and the level of vascular endothelial growth factor(VEGF)in co-cultured supernatant were evaluated by scratch assay,tube-formation assay,budding as-say and ELISA assay.Western blot was used to de-tect the expression of VEGFR2 and p-VEGFR2,and reverse verification was performed with inhibitors.A subcutaneous transplantation tumour model of hepatocellular carcinoma in nude mice was estab-lished,with Model group,Model+CBG group and Model+Apa group.The collagen expression of the transplantation tumour was observed by Masson staining,and the expression levels of vascular endo-thelial markers CD31 and CD34 were detected by immunofluorescence.RESULTS:When PLT:HUH7=200,the activity of HUH7 was the strongest,and the crosstalk between HUH7 and PLT significantly promoted the proliferation of EC(P＜0.01).Com-pared with Control group,the migration,tube-for-mation and budding ability of Crosstalk group were enhanced,and those of Intervention group were lower than those of Crosstalk group(P＜0.01).The expression level of VEGF in the supernatant of Crosstalk group was higher than that of Control group,while that of Intervention group was lower than that of Crosstalk group(P＜0.01).The expres-sion level of p-VEGFR2 protein in Crosstalk group was significantly higher than that of Control group,but the expression level of Intervention group was lower than that of Crosstalk group(P＜0.01).Large collagen fibre deposition was seen in the Model group,and CBG intervention significantly reduced collagen fibre deposition in the transplanted tu-mour tissues.CD31 and CD34 expression was pres-ent in the hepatocellular carcinoma transplanted tumour tissues in the Model group,and CBG inter-vention significantly reduced the expression of CD31 and CD34 in the liver cancer transplanted tu-mour tissues(P＜0.01).CONCLUSION:PLT enhances angiogenesis in hepatocellular carcinoma,and CBG may inhibit its tube-forming ability via the VEGF/VEGFR2 pathway.

Objective:To study the anatomy of the perforator propeller flap of superior lateral genicular artery, and to explore a surgical method and clinical application in repair of the soft tissue defect of anterolateral knee with the flap.Methods:From September 2019 to September 2021, 8 knees of 4 chilled fresh specimen of adults were studied. The perforators of the superior lateral genicular artery were observed. The length, outer diameter of the perforators, and the locations of the skin perforation were recorded. The superior lateral genicular artery perforator propeller flaps were then applied clinically to 5 patients(3 males and 2 females) with soft tissue defects of anterolateral knee. Two of the patients had combined ligament injury and(or) bone joint exposure. The age of patients ranged from 25 to 48 years old, at 33.4 years old in average. The sizes of soft tissue defects ranged from 4.0 cm×4.0 cm to 8.0 cm×5.0 cm. The sizes of perforator propeller flaps of superior lateral genicular artery were 10.0 cm×5.0 cm to 13.0 cm×6.0 cm. The superior perforating vessels of the superior lateral genicular artery were found and marked at the points of skin perforation. Preoperative contrast-enhanced ultrasound were performed to confirm the dominant perforating vessels and had the skin perforating points marked. Intraoperative CDU were further performed to confirm the points of dominant perforating vessels. Perforator propeller flaps were designed depending on the size of the anterolateral soft tissue defect, and flaps were prepared and transferred to the defect sites. Postoperative follow-ups were conducted at outpatient clinic. The survival of the flap and knee function were observed according to the Bai-ly knee scoring.Results:The anatomy showed that an average pedicle length of the superior lateral genicular artery perforator was(8.2±0.9) cm, with an average starting outer diameter at(1.1±0.2) mm. All 5 flaps survived during the follow-up that lasted for 10 to 24 months, with an average of 15.3 months. All flaps healed in 2 weeks after surgery without complications such as soft tissue infection, bone and joint infection were observed. At the last follow-up, no obvious bloated appearance of the flaps were observed. The colour and elasticity of the flaps were similar to the surrounding skin. The knee function was assessed: 4 patients were in excellent and 1 in good. The range of knee flexion and extension was from 100° to 150°. The patients were satisfied with the appearance and function of the knees.Conclusion:The size of the perforator of superior lateral genicular artery and the pedicle length are ideal. The propeller flap can be used to repair the soft tissue defect around the anterolateral knee, with satisfactory functional recovery of a knee. It is a good method to repair the soft tissue defect around anterolateral knee.

OBJECTIVE：To evaluate the economy performance of dexamethasone （DXM）combined with rituximab （RTX） for the first-line treatment of chronic primary immune thrombocytopenia （ITP）in adults. METHODS ：From the perspective of China ’s medical and health system ，Markov model for eight states was constructed with a period of 4 weeks and a time limit of 20 years， using DXM regimen as control. The cost-utility of DXM+RTX regimen for the treatment of chronic ITP in adults were evaluated. The parameters of clinical efficacy and utility value were derived from own published literature ；cost parameters were from the MENET website and the official websites of local health committees and medical insurance bureaus ；one-way sensitivity analysis ， probability sensitivity analysis and scenario analysis were performed to observe the uncertainty of model and data source. RESULTS：The average cost of DXM+RTX regimen was 51 064 dollars and that of DXM regimen was 50 455 dollars. Compared with DXM regimen ，DXM+RTX regimen yielded an additional 0.14 QALYs for each patient ；the incremental cost-effectiveness ratio（ICER）was 4 356 dollars/QALY，and was lower than the willingness-to-pay threshold of China ’s per capita gross domestic product（GDP）in 2020. In the one-way sensitivity analysis ，the cost of drugs was the main driver in the model. Probability sensitivity analysis demonstrated that DXM+RTX regimen had 57.5％-61.0％ probability of being cost-effective at a willingness- to-pay threshold of 1-3 times per capita GDP in 2020. The results of scenario analysis showed that DXM+RTX regimen would have obvious long-term benefits ，and the utility value had little impact on the conclusion. CONCLUSIONS ：DXM + RTX is more economical than DXM in the treatment of chronic ITP in adults ，but the results have the uncertainty.

Bladder cancer is a kind of urothelial cancer with high incidence and heterogeneity. From superficial bladder tumors to muscular invasive malignant tumors, due to their high-frequency recurrence and metastatic characteristics, they are inherently refractory. Although a comprehensive treatment with surgery as the main focus while chemotherapy, radiotherapy, and immunotherapy as a supplement has been formed, the limitations of chemotherapy regimens, the unpopularity of radiotherapy, and the low efficiency of immunotherapy, make clinical decision-making still difficult. Recently, a number of targeted therapies have emerged in bladder cancer and have shown good responses. Transient receptor potential (TRP) channel are novel therapeutic targets and current research hotspots in bladder cancer. This review discusses the anti-tumoral molecular mechanism of TRP channel in bladder cancer, its feasibility as a potential therapeutic target, and the prospects of drug combination to sensitize platinum-based chemotherapy.

PURPOSE: Effective biomarkers and models are needed to improve the prognostic prospects of clear cell renal cell carcinoma (ccRCC). The purpose of this work was to identify DNA methylation biomarkers and to evaluate the utility of DNA methylation analysis for ccRCC prognosis. MATERIALS AND METHODS: An overview of genome-wide methylation of ccRCC tissues derived from The Cancer Genome Atlas (TCGA) database was download for analysis. DNA methylation signatures were identified using Cox regression methods. The potential clinical significance of methylation biomarkers acting as a novel prognostic markers was analyzed using the Kaplan-Meier method and receiver operating characteristic (ROC) curves. RESULTS: This study analyzed data for 215 patients with information on 23171 DNA methylation sites and identified a two-DNA methylation signature (cg18034859, cg24199834) with the help of a step-wise multivariable Cox regression model. The area under the curve of ROCs for the two-DNA methylation signature was 0.819. The study samples were stratified into low- and high-risk classifications based on an optimal threshold, and the two groups showed markedly different survival rates. Moreover, the two-DNA methylation marker was suitable for patients of varying ages, sex, stages (I and IV), and histologic grade (G2). CONCLUSION: The two-DNA methylation signature was deemed to be a potential novel prognostic biomarker of use in increasing the accuracy of predicting overall survival of ccRCC patients.
Biomarkers ; Carcinoma, Renal Cell ; Classification ; DNA Methylation ; Genome ; Humans ; Methods ; Methylation ; Prognosis ; ROC Curve ; Survival Rate

Objective To evaluate the effect of living skin equivalents (LSE) constructed of mixed autologous and allogeneic skin cells and human amnion which served as a matrix on repairing scar contracture of the hand in a patient with recessive dystrophic epidermolysis bullosa (RDEB).Methods Skin tissues were obtained from a patient with RDEB and her mother,and epidermal keratinocytes and dermal fibroblasts were isolated from these tissues and cultured in vitro separately.Human amnion was obtained from the placenta of an unrelated healthy parturient undergoing cesarean delivery,and served as a matrix of the LSE.The autologous and allogeneic fibroblasts were mixed and seeded on the stromal side of the amnion,and then the autologous and allogeneic keratinocytes were mixed and seeded on the epithelial side of the amnion,so as to construct the human amnion-LSE (AM-LSE).Histological examination was performed to observe the structure of the skin tissues obtained from the patient and her mother,and immunofluorescence staining was conducted to detect the expression of type Ⅶ collagen in the skin tissues of the patient and her mother and in the AM-LSE.The AM-LSE was grafted on the skin defects of the patient after release of scar contracture of the hand.After grafting,the survival status of the AM-LSE graft and repairing effect on the wounds were evaluated.Results The constructed AM-LSE consisted of dermis,multilayered and fully differentiated epidermis and well-developed basement membrane.Immunofluorescence examination revealed that type Ⅶ collagen was linearly distributed along the basement membrane.Half a year after grafting,the AM-LSE survived well,and no obvious rejection reaction was observed.No blisters or ulcers occurred at the recipient sites,and the scar contracture was mild.The grafted area showed normal skin color with soft texture.The patient could grab and hold things,which had met self-care requirements of daily living.Conclusions The AM-LSE constructed of mixed autologous and allogeneic skin cells have good histological structures,and can be grafted on the wounds after resection of the scars in a RDEB patient.After grafting,no obvious rejection reaction was observed,and the skin was not liable to develop blisters,ulcers or scar contracture due to friction.

ObjectiveTo explore the effect of percutaneous locking compression plate(LCP) internal fixation for distal tibial comminuted fracture.Methods35 patients of distal tibial Comminuted fracture were treated with percutaneous locking compression plate(LCP) internal fixation.ResultsAll patients were followed up for average 1.8 years(ranging from 1 to 2.5 years).All of cases have healed well,and the average bone healing was 14.6 weeks (ranging from 8 to 28 weeks ).All of them have no infection and no loosening or breakage of internal fixation.According to Mazur criterion,excellent effect 17 cases,good effect 16 cases,fair effect 1 case poor effect 1 case,94.3％ was excellent or good.ConclusionThe LCP internal fixation for distal tibial comminuted fracture have the advantage of less invasive,good internal fixation and bone union fastly and little complications.

The effects of RNAi-mediated gene silencing of LRIG3 expression on cell cycle and survival of human glioma cell line GLI 5 and the possible mechanisms were explored.The plasmids pGenesil2-LRIG3-shRNA l and pGenesil2-LRIG3-shRNA2 were transfected into GL 15 glioma cells respectively by using Metafectine,and the transfected cells that stably suppressed LRIG3 expression were selected by G418.The control cells were transfected with negative control shRNA.The changes in LRIG3 mRNA and protein levels were measured by RT-PCR and Western blot.The apoptosis rate and cell cycle were analyzed by flow cytometry.As compared with the negative shRNA-transfected GL15 cells,LRIG3 mRNA expression in GLI5 cells transfected with pGenesil2-LRIG3-shRNAl and pGenesil2-LRIG3-shRNA2 was silenced by 52.4%,63.8%,and LRIG3 protein expression was re-duced by 50.9% and 67.4% respectively.The LRIG3-specific siRNA transfected cells had higher proliferation rate than control cells.Cell cycle analysis showed that silencing LRIG3 increased the percentage of G2/M phase cells and the proliferation index significantly (P<0.01).Silencing LRIG3 could inhibit the apoptosis of GLl5 cells (P<0.05).These findings suggest that the siRNA targeting LRIG3 gene shows a dramatic inhibitory effect on RNA transcription and protein expression,then promoting the proliferation of GLI5 cells,arresting GLI5 cells in G2/M phase,and suppressing apoptosis ofGL15 cells.

Objective To study the reversal effect of tetrandrine (TET) on the multidrug resistance of Hep-2/ADM cell line and the possible mechanisms. Methods MTT method was used for the detection of the inhibitory effect of different concentrations of TET on the multidrug resistance of Hep-2/ADM cell line for selection of the non-toxicity concentration. MTT reduction assay was used to investigate the drug interaction of TET and vincristine (VCR) on HNE-1 (200) cell line. The reversal index (RI), accumulation and excretion of drug, and the apoptosis of Hep-2/ADM cell line were detected by flow cytometry. Results No toxicity to Hep-2/ADM cell line was found when TET concentration was lower than 1.5 ?g/ml. After interaction with TET (1.0 ?g/ml and 1.5 ?g/ml), RIs of VCR on Hep-2/ADM were 1.72 and 2.00 folds, respectively. TET at the concentration of 1.5 ?g/ml could increase the drug excretion and accumulation and also the apoptotic rate of VCR-induced Hep-2/ADM cell line. Conclusion TET can reverse the multidrug resistance of Hep-2/ADM cell line. The reversal effect may be related to the concentrations of TET. The possible mechanism may be that TET can increase the cell apoptosis of VCR-induced Hep-2/ADM cell line and can also increase the drug excretion and accumulation of the multidrug resistance cell line.

Objective To study the effect of different extraction parts from Xiongjiang Decoction(XD) on promoting uterine muscle contraction(UMC).Methods UMC in normal mice was measured in vitro by applying serologic pharmacological test.Results Alcohol extracts of XD had an obvious effect on promoting UMC,but the volatile oil and water-extraction alcohol-precipitation extracts had an inhibitory effect.Chloroform part of alcohol extracts had a stronger effect on UMC than other extraction parts.Conclusion The different parts or components extracted from the same decoction have different effects.