Background and purpose:Colorectal cancer(CRC)is one of the major malignant tumors threatening human health worldwide,with long-term high incidence and mortality rate.Potassium channel modulatory factor 1(KCMF1)is a member of the E3 ubiquitin ligase family.It binds to target proteins through the RING domain and participates in the regulation of a variety of biological processes in vivo.However,the function of KCMF1 in CRC remains unclear.This study aimed to investigate the expression level of E3 ubiquitin ligase KCMF1 in colorectal tumor,and to explore the effects of KCMF1 on the proliferation of CRC cells and its underlying molecular mechanism.Methods:The The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases were used to analyze the expression level of KCMF1 in CRC tissues and adjacent tissues and the association between the KCMF1 expression and the prognosis of CRC patients.Furthermore,immunohistochemical staining was performed to detect the protein level of KCMF1 in 90 paired human CRC tissues and adjacent non-tumor tissues.Lentiviral shRNA delivery system was employed to specifically target the KCMF1 gene(shKCMF1)in HCT116 and HCT15 CRC cell lines.The effects of KCMF1 knockdown on cell proliferation,apoptosis and cell cycle distribution were assessed by methyl thiazoyl terazolium(MTT)assay,colony formation assay,Western blot and flow cytometry.Changes in the transcriptional profile in HCT116 cells upon KCMF1 knockdown were identified by RNA sequencing(RNA-Seq),and the affected signaling pathways were evaluated by bioinformatics analysis.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot,luciferase reporter assay and cell immunofluorescence assay were utilized to validate the alteration of the affected signaling pathway.Results:The TCGA and GTEx databases and IHC results showed that the mRNA and protein expression levels of KCMF1 in CRC tissues were significantly upregulated compared with adjacent tissues(P＜0.01).KCMF1 expression level was negatively correlated with the survival time of patients with CRC(P＜0.01),and was positively associated with CRC clinical stage(P＜0.05).Compared with control cells,KCMF1 knockdown significantly inhibited the proliferation of HCT116 and HCT15 cells(P＜0.001),induced cell apoptosis(P＜0.001),and led to cell cycle arrest in G1 phase(P＜0.01).RNA-Seq analysis showed that KCMF1 was involved in the regulation of several signaling pathways,including nuclear factor-κB(NF-κB)signaling pathway.KCMF1 knockdown reduced the transcription levels of the target genes of NF-κB signaling pathway,including BCL-XL,XIAP and CIAP(P＜0.05),and suppressed the expression of phosphorylated p65 and nuclear translocation of p65(P＜0.01).Meanwhile,the activity of NF-κB reporter was reduced in tumor cells upon KCMF1 knockdown(P＜0.01).Conclusion:The expression of KCMF1 is significantly upregulated in human CRC tissues and positively associated with advanced clinical stage and poor prognosis.KCMF1 may promote the proliferation of CRC cells by activating the NF-κB signaling pathway.KCMF1 may be a potential new therapeutic target for CRC.

Objective:To investigate the effects of fasudil hydrochloride(FH) on Rho-associated kinase 2(ROCK2) protein and ferroptosis in hippocampal area during early brain injury in rats with subarachnoid hemorrhage(SAH).Methods:Total 36 SPF grade Sprague-Dawley rats were divided into 3 groups by random number table method: Sham group, SAH group and SAH+ FH (a ROCK2 protein inhibitor) group (FH goup) with 12 rats in each group.SAH animal model was established by internal carotid artery perforation.The rats in FH group were injected intraperitoneally with FH(15 mg/kg) 30 minutes after successful modeling, and rats in Sham group and SAH group were injected intraperitoneally with the same volume of 0.9% sodium chloride solution.Twenty-four hours after the intervention, shuttle box test was used to observe the learning and memory ability of rats.The Fe 2+ content in rat hippocampus tissue was detected by colorimetry, and the protein levels of ROCK2 and ferroptosis-related long-chain acyl-CoA synthetase 4(ACSL4) and glutathione peroxidase 4(GPX4) in hippocampus were detected by immunohistochemistry and Western blot.Statistical analysis was performed by SPSS 20.0 software.One-way ANOVA was used for multigroup comparison, and LSD test was used for further pairwise comparison. Results:(1)In the shuttle box test, there were statistically significant differences in the number of avoidance reactions and avoidance reaction time of rats among the three groups( F=20.348, 22.316, both P<0.05). The number of avoidance reaction in SAH group was less than that in Sham group ((17.92±2.94) times, (27.13±3.48) times, P<0.05), the time of avoidance reaction in SAH group was longer than that in Sham group ((9.15±2.87) s, (3.68±1.09) s, P<0.05), while the number of avoidance reaction in FH group ((21.63±4.11) times) was more than that in SAH group, and the time of avoidance reaction ((6.08±1.76) s) was shorter than that in SAH group (both P<0.05). (2) The colorimetry results showed that there was a statistically significant difference in the content of Fe 2+ in hippocampus of rats among the three groups( F=7.965, P<0.05). The Fe 2+ content in SAH group was significantly higher than that of Sham group((0.091±0.032) nmol/mg, (0.038±0.024) nmol/mg, P<0.05), and the Fe 2+ content in the FH group ((0.065±0.021) nmol/mg) was lower than that of SAH group ( P<0.05). (3) There were significant differences in the number of ROCK2, ACSL4 and GPX4 positive cells in hippocampus of rats among the three groups in immunohistochemistry ( F=7.602, 14.171, 36.077, all P<0.05). The positive cells of ROCK2 and ACSL4 in SAH group ((21.63±4.72), (55.13±19.41)) were significantly higher than those of Sham group ((11.63±3.62), (23.38±3.74)) (both P<0.05), and the positive cells of ROCK2 and ACSL4 in FH group ((15.88±6.64), (44.75±8.29)) were both lower than those of SAH group(both P<0.05), while the number of GPX4 positive cells in SAH group (25.38±6.30) was significantly lower than that of Sham group (60.25±10.36) ( P<0.05), and the number of GPX4 positive cells in FH group (45.13±7.51) was higher than that of SAH group( P<0.05). (4)The results of Western blot showed that there were significant differences in the expression levels of ROCK2, ACSL4 and GPX4 proteins in the hippocampus of rats among the three groups( F=4.812, 12.573, 10.849, all P<0.05). The protein expression levels of ROCK2 and ACSL4 in SAH group were significantly higher than those in Sham group(both P<0.05), and the protein expression levels of ROCK2 and ACSL4 in FH group were lower than those in SAH group (both P<0.05), while the expression level of GPX4 protein in SAH group (0.27±0.09) was significantly lower than that in Sham group( P<0.05), and the expression level of GPX4 protein in FH group was higher than that of SAH group ( P<0.05). Conclusion:FH can inhibit ferroptosis in the hippocampus and improve the learning and memory ability of rats, and the mechanism may be related with down-regulation of ROCK2 protein.

Objective:To analyze the commonalities and differences of " Internet plus healthcare" related policies in the eastern, central and western regions of China before and after the COVID-19 pandemic and to provide references for further improving China′s " Internet plus healthcare" policy.Methods:Collecting the policies of " Internet plus healthcare" promulgated by 31 provinces(autonomous regions, municipalities)in China from April 2018 to December 2020 on the websites of various government departments, extracting keywords based on content analysis methods, and calculating degree centralities using UCINET 6 software. A comparison of the policies of the eastern, central and western regions between the pre epidemic stage(April 2018 to December 2019)and the COVID-19 pandemic stage(January 2020 to December 2020)was carried out.Results:In the pre epidemic stage, Internet diagnosis and treatment, " Internet plus medical service price" and quality supervision were the focus of attention in all regions. But the eastern region paid more attention to the policy planning of Internet hospital operation and Internet medical application, the central region focused on the transformation and efficiency improvement of medical service process by Internet technology, and the western region paid more attention to encourage the application of telemedicine to solve the difficulty of seeing a doctor. In the pandemic stage, the " Internet plus service price" was the policy focus in all regions, mostly in medical insurance payment, and the policy difference was not obvious among the eastern, central and western regions.Conclusions:Before and after the COVID-19 pandemic, the policy of " Internet plus healthcare " promulgated by the local governments in eastern, central and western regions had different emphasis and different characteristics. Our suggestion was to improve the policy system, standardize industry supervision, strengthen information security, orderly interconnect medical insurance, and actively promote the exchange of policy experiences among the provinces.

The crown-root ratio (C/R) theory of natural teeth has been widely recognized in the field of stomatology,and has important clinical significance in predicting and assessing the prognosis of natural teeth as well as for abutment selection during denture restoration. In the past few decades, scholars have advocated for the implantation of implants as long in length as possible to improve the success rate according to the theory of crown-root ratio of natural teeth. However, with the application of short implants, our philosophy of implantation has changed, and the relationship between the crown-implant (C/I) ratio and complications has become one of the current research hotspots. In this paper, the concept of the crown-implant ratio, the research progress of the C/I ratio, the implant survival rate and clinical complications of implant restoration were reviewed and summarized, and the following suggestions were put forward: although most studies have shown no significant correlation between the C/I ratio and implant survival or marginal bone loss, this relationship may increase the risk of mechanical complications. A C/I < 3 and a crown length < 15 mm are recommended in implant restoration; when ultra-short implants are applied, the implant system can increase the bone-to-implant contact area, and splint prostheses such as crown or bridge are recommended.

Objective : To explore the clinical effect of the alveolar crest approach in the treatment of odontogenic maxillary sinusitis and the repair of edentulous implants in this area. Methods: This was a retrospective case series of 20 patients with odontogenic sinusitis. The pathogenesis in each case was investigated. After elimination of the dental origin, each patient was treated with flushing, drainage and anti-inflammatories through the alveolar crest approach. Postoperative CBCT reexamination was performed to evaluate the therapeutic effect. Maxillary sinus elevation surgery with simultaneous or delayed implantation was performed after maxillary sinusitis healing was confirmed. The patients were followed postoperatively. Results : Twenty patients with odontogenic sinusitis were treated by the alveolar crest approach, and 17 were cured, for a cure rate of 85%. Among them, 17 of the maxillary sinusitis patients were followed for 1 year, with good results. Conclusion The alveolar crest approach is feasible for the treatment of odontogenic maxillary sinusitis and can serve as a minimally invasive method for the repair of edentulism in this area and implantation.

Objective : To investigate the effect of a laser-etched pure titanium surface on proliferation of the human osteosarcoma cell line MG63 and to provide a basis for study of implant surface modification. Methods: The pure titanium plate was cut into titanium pieces by a numerical control machine tool and divided into smooth surface and laser etching groups. The titanium surface of the laser etching group was etched with an Nd:YAG continuous wave laser using predetermined parameters, and the surfaces were observed by scanning electron microscopy (SEM). The surface micromorphology of each titanium sheet was evaluated. The relative element content of the titanium surface was measured by energy dispersive X-ray spectroscopy (EDS). The Ra value of each surface was determined using the Veeco roughness tester. MG63 cells were inoculated on 2 sets of titanium tablets. At 1, 3, and 6 h postinoculation, cell adhesion to the two groups of titanium sheets was observed under the microscope. At 24 h after inoculation, cellular F-actin was directly stained using immunofluorescence, and the morphology of the cytoskeleton was observed by laser confocal microscopy. Cell proliferation was examined at 1, 3, and 5 d using a MTS kit, and the data were analyzed with SAS 9.4. Results : The surface of the smooth surface group was smooth and flat, the element composition was pure titanium, and the roughness Ra was 179.23 nm. The surface of the laser-etched group formed a regular and uniform pore structure. The composition was mainly Ti, O, C, etc, and the surface roughness Ra was 14.11 μm. A large number of cells were uniformly distributed on the two titanium sheets in the observations at 1, 3, and 6 h. At 24 h postinoculation, MG63 cells were completely stretched on the two sets of titanium sheets and had extended a large number of pseudopods and microfilaments to cross-link with peripheral cells; moreover, the cell division phase was observed. The cell proliferation of the two groups at 1, 3, and 5 d showed a significant increase with time, indicating that no cytotoxicity occurred on the surfaces of the two groups. However, the cell proliferation in the laser-etched group was superior to that in the mechanical smooth surface group. Conclusion The surface morphology of titanium can be controlled by laser etching, which is conductive to increase the microstructure of implants without cytotoxicity and promoting osteoblast proliferation in the early stage.

Objective To evaluate the therapeutic outcome of horizontal bone augmentation by using the modified split crest technique of piezosurgery in mandible molar area. Methods The present study reviewed the record of 15 patients from 2016 January to 2017 March in the department of oral implantology center of Stomato-logical Hospital of Southern Medical University,who underwent the modified split crest technique of piezosurgery in mandible molar area. Preoperative CBCT was performed to determine the bone thickness of the toothless alveo-lar crest of the mandibular posterior region,and the bone thickness was less than 3 mm. The modified split crest technique of piezosurgery was performed for the horizontal bone augmentation at mandibular posterior region. Meanwhile,GBR and the postoperative CBCT were performed to determine the bone width increments,followed with 1-week anti-inflammatory therapy postoperation. Three months after the operations,the bone width incre-ment was examined by CBCT. SPSS13.0 software was used to analyze the collected data. Results The modified split crest technique of piezosurgery in mandible molar area was successfully established,resulting in the mini-mal operative wounds and the slight postoperative reaction,without obvious infection. The postoperative crest width increase achieved 6.05 ± 0.65 mm. At 3 months after operation,the crest width increase reduced to(5.81 ± 0.61)mm. There were significant differences of the alveolar ridge thickness between the preoperative and post-operative patients(P<0.05). There were significant differences between the preoperative crest width and the 3 months after treatment(P<0.05). Conclusion Horizontal bone augmentation of the modified split crest tech-nique of piezosurgery was efficient,which could be performed in very narrow alveolar ridge,and which was rela-tively safe and accurate,with less trauma. No damage was observed to the soft tissue,and the cutting direction can be well controlled.

Objective To investigate the impacts of LASP-1 on the behavior and cell proliferation ability of tumor derived osteoblast-like cells and non tumor derived osteoblasts. Methods QRT-PCR and Western blot were performed to detect LASP-1 expressions in mouse bone marrow mesenchymal stem cells(BMMSC),human osteoblasts (hOB) and human osteosarcoma cell (MG-63). With the construction of recombinant vector and the transfection of shLASP-1,the expression changes in both LASP-1 and osteocalcin(OCN),and the cell number of cell-cycle phase distribution were analyzed. Results With up-or down-regulation of LASP-1,osteocalcin(OCN) expression,cell proliferation ability and the cell number of G2/M phase in non tumor cells(BMMSC,hOB)had no obvious changes. However,as for the tumor cells(MG-63),with an increased expression of LASP-1,the expression of OCN,cell proliferation number and the cell number at G2/M phase changed. There was no significant effect on the moving ability of each cell. Conclusions LASP-1 has no significant effect on the osteogenic proper-ties,cell proliferation ability and cell cycle distribution of non tumor derived cells,but has a regulatory effect on tumor derived cells(MG-63).

Objective: To investigate the effect of pure titanium surface of large diameter TiO2 nanotubes modified by RGD peptide on the adhesion and proliferation of MG63 osteoblasts. to provide theoretical proof for developing titanium implants. Methods: Commercially pure titanium discs were divided into four groups and treated with SLA to obtain a microrough surface (SLA group). Then, nanotubes were imposed on this microrough surface by anodization (SLA+80 group). The surface was then modified by dopamine (DOPA) (DOPA Group), after which bioactive RGD peptide layers were generated on the TiO2 nanotube surfaces via electrochemical and molecular self-assembly techniques (RGD group). The titanium surface morphology and elemental composition of each group were characterized by field emission scanning electron microscopy (FE-SEM) and X-ray photoelectron spectroscopy (XPS). MG63 cells were cultured in vitro to evaluate biological activities of titanium before and after treatment, including the evaluation of early-stage cell adhesion capacity by fluorescence microscopy, proliferation capacity by MTS assay, and mRNA expression of the cell osteoblast-related genes alkaline phosphatase (ALP) and osteocalcin (OCN) by qRT-PCR. Results: FE-SEM and XPS showed that hierarchical micro/nanosurfaces decorated with TiO2 nanotubes were produced on titanium using sandblasting and large grit etching combined with anodization, dopamine was then self-polymerized to form a polydopamine film on the TiO2 nanotube surfaces, and RGD peptides were then conjugated to the polydopamine film, finally forming RGD peptide-modified bioactive layers. In vitro experiments showed that compared with the other three materials, the RGD-modified material was more conducive to cell adhesion and proliferation (P < 0.05). The expression levels of ALP and OCN mRNA in the RGD group were significantly higher than those in the SLA group and DOPA group (P < 0.05). Conclusion Hierarchical micro/nanosurfaces decorated with TiO2 nanotubes functionally modified with RGD peptides have good biocompatibility and could be used for developing titanium implants and further improving early osseointegration.

Objective To evaluate clinical effect of Compound Fanshiliu Recipe(CFR) for the treatment of over-weighted type 2 diabetes mellitus(T2DM). Methods One hundred and twenty over-weighted T2DM patients were randomized into treatment group and control group, 60 cases in each group. Both groups were given oral use of Metformin Hydrochloride Enteric-Coated Tablets, and additionally the treatment group was treated with CFR orally. The treatment of both groups lasted for 12 continuous weeks. Before and after treatment, we detected the waist circumference (WC), body mass index (BMI), and the levels of fasting blood glucose(FBG), postprandial 2-hour blood glucose(P2hBG), glycosylated hemoglobin(HbA1c), total cholesterol(TC), triglyceride (TG), and free fatty acid (FFA). The therapeutic effect of the two groups was also evaluated after treatment. Results (1) The total effective rate for traditional Chinese medical symptoms was 95.00% in the treatment group, and was 70.00%in the control group, the difference being significant(P<0.01).(2) After treatment, WC and BMI were decreased in the treatment group(P<0.01 compared with those before treatment), but had no significant decrease in the control group(P>0.05). The decrease of WC and BMI of the treatment group was superior to that of the control group(P<0.05).(3) After treatment, levels of FBG, P2hBG, HbA1c, TC, TG, and FFA of both groups were significantly improved (P<0.05), and the improvement of the treatment group was superior to that of the control group(P<0.05).(4) Three patients of the treatment group suffered from slight gastric discomfort, and the discomfort was relieved without any special treatment. Other patients of the two groups had no adverse reaction. Conclusion Treatment of CFR combined with Metformin has been proved to be an effective and safe approach to decrease blood glucose and blood lipid levels, promote weight loss, and ameliorate traditional Chinese medical symptoms.