Objective:To investigate the number of necroptotic renal tubular epithelial cells in renal tissues of patients with chronic kidney disease (CKD) and the correlation with clinicopathologic parameters, and explore its role in the progression of the excessive loss of renal tubular cells and chronic kidney injury.Methods:Renal tissue samples from 60 patients (18-65 years old) with CKD proven by kidney biopsy in the First Affiliated Hospital of Hainan Medical University from June 2017 to June 2019 were collected. According to internationally accepted K/DOQI guidelines, the patients were divided into 1-4 stages of CKD, with 15 cases in each stage. The number of necroptotic renal tubular epithelial cells in patients with different stages of CKD was detected using receptor-interacting protein 3 (RIP3) and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) fluorescent staining, and the expression of RIP3 and MLKL, marker protein of necroptosis, was detected by immunohistochemistry. Pearson correlation analysis was used to analyze the correlation between the percentage of necroptotic renal tubular epithelial cells and clinicopathologic parameters. In addition, the expression of angiotensinogen Ⅱ receptor (AT2R) in renal tissue and its correlation with the percentage of necroptotic renal tubular epithelial cells were analyzed.Results:With the development of CKD, the structural destruction of renal tubules in patients with CKD was gradually aggravated, and the renal tubules in the corresponding areas were atrophied, accompanied by worsening interstitial fibrosis. The adjacent renal tubules were focally dilated and numerous protein tubules were seen in the tubules. Importantly, renal tubular injury score in second and third stage of CKD was significantly higher than that in control group (both P<0.01). TUNEL+RIP3 immunofluorescence staining results showed that the percentage of TUNEL/RIP3 double positive renal tubular epithelial cells (necroptotic renal tubular epithelial cells) in renal tubules of the second and third stage of CKD was higher (all P<0.01). Immunohistochemical results showed that RIP3, MLKL and AT2R proteins were mainly expressed in cytoplasm of renal tubular epithelial cells, and the expression of RIP3, MLKL and AT2R in renal tubular epithelial cells was higher in the second and third stage of CKD patients (all P<0.05). Pearson correlation analysis showed that the percentage of necroptotic renal tubular epithelial cells was positively correlated with blood urea nitrogen ( r=0.514, P=0.003), serum creatinine ( r=0.507, P=0.019), serum cystatin C ( r=0.571, P=0.026), serum uric acid ( r=0.592, P=0.008), renal tubules injury score ( r=0.901, P<0.001), renal interstitial fibrosis index ( r=0.700, P=0.001) and the expression of AT2R protein in renal tissue ( r=0.715, P=0.001). Conclusions:As CKD progresses, necroptosis of renal tubular epithelial cells in CKD patients occurs. The necroptotic cell death may be an important factor leading to renal tubular epithelial cell excessive death and the progression of chronic kidney injury. Furthermore, necroptosis of renal tubular epithelial cells may be related to the high expression of AT2R in kidney tissue.

[ ABSTRACT] AIM: To explore whether autophagy is involved in the excessive death of renal tubular epithelial cells in subtotal nephrectomy ( SNx) rats and the relationship between autophagy and necroptosis in the kidney of SNx rats. METHODS:Male Sprague-Dawley rats were randomly assigned to control group ( n=6 ) and SNx group ( n=42 ) .The rats in SNx group were subjected to SNx.Sham surgery was performed in the rats in control group.The rats in SNx group were divided into subgroups at 0, 4, 8 and 12 weeks ( n=6) and the other rats in SNx group were divided into SNx+vehi-cle group, SNx+necrostatin-1 (Nec-1) group and SNx+3-methyladenine (3-MA) group.The expression of RIP1, RIP3, LC3 and beclin-1 at mRNA and protein levels was measured at 0, 4, 8 and 12 weeks by qPCR and immunohistochemistry. The effects of Nec-1 or 3-MA on the protein expression of LC3-I, LC3-II and beclin-1, and production of reactive oxygen species ( ROS) in the rat kidney were determined by Western blot and DCFH-DA staining.The death of renal tubular epi-thelial cells in the SNx rats was observed by TUNEL staining and electron microscopy.Finally, the effects of Nec-1 and 3-MA on blood urea nitrogen ( BUN) , serum creatinine ( SCr) and the pathological changes of the renal tissues were ana-lyzed.RESULTS:The highest mRNA and protein levels of RIP1, RIP3, LC3 and beclin-1 appeared at the 8th week after SNx (P<0.01).Compared with the rats in SNx+vehicle group, the protein over-expression of LC3-II/I and beclin-1, re-nal tubular epithelial cells with typical morphological features of necroptotic cell death and TUNEL-positive renal tubular
cells were decreased in the SNx rats treated with Nec-1 and 3-MA (P<0.01), but 3-MA did not reduce the increased con-centration of ROS.In addition, treatment with Nec-1 and 3-MA obviously reduced BUN, SCr (P<0.05), glomeruloscle-rosis index and tubulointerstitial injury score (P<0.01).CONCLUSION:Autophagy participates in the excessive death of renal tubular epithelial cells in SNx rats.Inhibition of autograph prevents necroptotic cell death of renal tubular cells, and alleviates chronic renal injury in SNx rats.

Objective:To explore the efficacy of Ortho-Bridger System (OBS) for the treatment of complex tibial fractures.Methods:A retrospective study was conducted of the 64 patients with complex tibial fracture who had been treated at Department of Orthopaedics, Tongling People's Hospital from June 2014 to June 2018 using OBS. They were 46 males and 18 females, aged from 22 to 65 years (average, 44.3 years). There were 38 close comminuted or multi-segmental fractures complicated with skin and soft tissue defects and 26 cases of tibial exposure. The interval from injury to surgery ranged from 3 to 20 days (average, 13.6 days). The therapeutic outcomes were evaluated in terms of fracture union time, healing of skin and soft tissue defects and postoperative functional recovery.Results:The 64 patients were followed up for 12 to 50 months (average, 26.3 months). Obvious callus formation started at 3 to 8 months after operation and bony union was achieved at 9 to 18 months after operation. No obvious limitation was observed in the range of motion of the ankle and knee joints. Skin defects healed and completely covered the bone at 3 to 6 weeks after operation.Conclusion:OBS can lead to fine clinical efficacy in the treatment of complex tibial fractures, like open and comminuted multi-segmental ones complicated with soft tissue defects.

Objective To evaluate the prevalence of chronic kidney disease (CKD) in Chinese adult health check-up population,and to compare with the prevalence of CKD in the study of the general population as well as the large CKD cross-sectional study in China.Methods Epidemiological studies about CKD in Chinese adults health check-up population from January 2007 to December 2017 were searched in PubMed,SinoMed,CNKI,VIP and Wanfang Data.Meta-analysis of the prevalence of CKD was performed with software of Stata 12.0.Subgroup analyses of CKD staging,urban and rural,as well as geographical areas of the general population were executed.Results Twenty-two studies from adult health check-up population were included (238 349 persons).Egger's regression showed no publication bias (P ＞ 0.05).The unstandardized prevalence rate of CKD was 12.49％ (male 12.8％,female 12.5％).The respective unstandardized prevalences of proteinuria,hematuria and eGFR decline were 5.90％,5.83％ and 2.75％.The unstandardized prevalences of CKD in urban and rural population were 13.21％ and 11.90％.The stages of CKD were mainly concentrated in the early stages.There was no significant difference in the non-standard detection rate of total eGFR decline among the adult medical examination population,the general population and the population studied cross-sectionally (P ＞ 0.05).Furthermore,no significant difference in the non-standard detection rate of total hematuria and male hematuria was found between the adult health check-up population and the general population.In addition,the total proteinuric non-standard detection rate of the adult general population was similar with that of population studied cross-sectionally (P ＞ 0.05).Conclusions The prevalence of CKD in Chinese adults is higher,the overall prevalence is however underestimated.The results of epidemiological investigation in adult health check-up population are similar to those of the general population,especially in men.

Objective: To evaluate the value of Kirschner wire internal fixation in the treatment of sagittal mandibular condylar fractures. .Methods : From January 2019 to January 2020, 13 patients (19 sides) with mandibular condylar sagittal fracture treated by Kirschner wire internal fixation at the Stomatological Medical Center, Foshan Hospital of Traditional Chinese Medicine were retrospectively analyzed. After conventional surgical incision and exposure and reduction of the mandibular condyle, 2-4 Kirschner wires were used for fixation, and other maxillofacial fractures were treated simultaneously. The reduction accuracy and stability of the free mandibular condyle were evaluated by CBCT one week after the operation, and the occlusion relationship, opening degree and opening type were evaluated by clinical examination. Results: All patients had good fracture alignment and no twisting, breaking or loosening of the Kirschner wire. The occlusion relationship, opening degree and opening shape recovered well after the operation. Conclusion Kirschner wire is effective in treating sagittal fractures of mandibular condyles.

Purpose: Numerous studies have shown that the frequency of myeloid-derived suppressor cells (MDSCs) is associated with tumor progression, metastasis, and recurrence. Chemokine (C-C motif) ligand 3 (CCL3) may be secreted by tumor cells and attract MDSCs into the tumor microenvironment. In the present study, we aimed to explore the molecular mechanisms whereby CCL3 is involved in the interaction of breast cancer cells and MDSCs. Methods: The expression of CCL3 and its receptors was investigated using real-time polymerase chain reaction, western blotting, and enzyme-linked immunosorbent assay. The cell counting Kit-8, wound healing, and transwell assays were performed to study cell growth, migration, and invasion. Cell cycling, apoptosis, and the frequency of MDSCs were investigated through flow cytometry. Transwell assays were used for co-culture and chemotaxis detection. Markers of the epithelial-mesenchymal transition (EMT) were determined with western blotting. The role of CCL3 in vivo was studied via tumor xenograft experiments. Results: CCL3 promoted cell proliferation, migration, invasion, and cycling, and inhibited apoptosis of breast cancer cells in vitro. Blocking CCL3 in vivo inhibited tumor growth and metastases. The frequency of MDSCs in patients with breast cancer was higher than that in healthy donors. Additionally, MDSCs might be recruited by CCL3. Co-culture with MDSCs activated the phosphoinositide 3-kinase-protein kinase B-mammalian target of rapamycin (PI3K-Akt-mTOR) pathway and promoted the EMT in breast cancer cells, and their proliferation, migration, and invasion significantly increased. These changes were not observed when breast cancer cells with CCL3 knockdown were co-cultured with MDSCs. Conclusion CCL3 promoted the growth of breast cancer cells, and MDSCs recruited by CCL3 interacted with these cells and then activated the PI3K-Akt-mTOR pathway, which led to EMT and promoted the migration and invasion of the cells.

Objective : To explore the effect of negative pressure sealing drainage on the treatment of maxillofacial-neck-mediastinal infection in multiple spaces. Methods: Vacuum sealing drainage (VSD) was applied in five patients with maxillofacial-neck-mediastinal infection caused by odontogenic infection accompanied by diabetes or renal failure and other systemic diseases. After extensive debridement, a negative pressure drainage sponge was placed in the pus cavity and then the wound was closed. Continuous negative pressure drainage was continued after the operation. At the same time, multidisciplinary consultation was applied to control basic diseases and, strengthen anti-inflammatory responses, and nutrition and other systemic treatments were applied. Results: Four patients underwent continuous negative pressure drainage and successful removal of the negative pressure sponge after inflammatory symptoms subsided. One patient′s inflammatory symptoms became more serious after the operation, and we performed another operation to change the placement of the negative pressure sponge. All 5 patients underwent VSD with negative pressure sponge replacement ranging from 1 to 3 times during treatment. After multidisciplinary consultation, they were all cured and discharged from the hospital. Conclusion For infection of the mediastinum, maxillofacial region and neck, local treatment and systemic treatment are emphasized, as well as the treatment of infected lesions and basic diseases. Negative pressure closure and drainage technology promotes the alleviation of inflammation, and multidisciplinary combined treatment is beneficial for the control of basic diseases.

Pyroptosis is the process of inflammatory cell death. The primary function of pyroptosis is to induce strong inflammatory responses that defend the host against microbe infection. Excessive pyroptosis, however, leads to several inflammatory diseases, including sepsis and autoimmune disorders. Pyroptosis can be canonical or noncanonical. Upon microbe infection, the canonical pathway responds to pathogen-associated molecular patterns (PAMPs) and damage-associated molecular patterns (DAMPs), while the noncanonical pathway responds to intracellular lipopolysaccharides (LPS) of Gram-negative bacteria. The last step of pyroptosis requires the cleavage of gasdermin D (GsdmD) at D275 (numbering after human GSDMD) into N- and C-termini by caspase 1 in the canonical pathway and caspase 4/5/11 (caspase 4/5 in humans, caspase 11 in mice) in the noncanonical pathway. Upon cleavage, the N-terminus of GsdmD (GsdmD-N) forms a transmembrane pore that releases cytokines such as IL-1

【Objective】 To explore the regulation of DIRAS2 gene expression by acetylated STAT3 and its involvement in the proliferation of triple-negative breast cancer (TNBC) cells. 【Methods】 The expression levels of DIRAS2 and acetylated STAT3 in TNBC tissues and cells were analyzed by database query, Western blotting, and qRT-PCR. TNBC cell lines MDA-MB-231 and SUM159 were selected, and lentivirus or plasmid was used to construct DIRAS2 overexpression and STAT3 wild or Lys685 mutation cell lines. The CCK-8 assay was used to evaluate the effect of DIRAS2 and STAT3 acetylation on the proliferation of TNBC cells. Western blotting, pyrosequencing, ChIP and IP were employed to investigate the regulatory effect and mechanism of acetylated STAT3 on DIRAS2 expression. 【Results】 The expression of DIRAS2 was decreased in TNBC tissues and cells. Pyrosequencing analysis found that the methylation level of CpG islands in the DIRAS2 promoter was increased in TNBC cells compared with normal breast epithelial cells, which promoted the growth of cancer cells. Furthermore, TNBC cells showed an increase in STAT3 acetylation, which was accompanied by a shift in the methylation status of the DIRAS2 promoter. ChIP and IP experiments showed that acetylated STAT3 could bind to the DIRAS2 promoter, and the STAT3 Lys685 mutation disrupted the interaction between STAT3 and DNMT1. 【Conclusion】 Acetylated STAT3 induces DIRAS2 promoter methylation by recruiting DNMT1, leading to loss of DIRAS2 expression and cancer cell proliferation in TNBC.