OBJECTIVE:The optimal non-surgical therapy for chronic ankle instability remains unclear due to the continuous introduction of novel treatment methods despite the availability of several non-surgical options for improving foot and ankle function and dynamic balance in chronic ankle instability patients.This study aims to investigate the most effective non-surgical therapy options to improve foot and ankle function and dynamic balance for patients with chronic ankle instability using a network meta-analysis. METHODS:Using"CAI,exercise,and randomized controlled trial"as search terms,a literature search of PubMed,Embase,Cochrane Library,and Web of Science databases was conducted through a computer network to collect information from the databases from their inception to March 2024 on non-surgical therapies for the treatment of chronic ankle instability randomized controlled trials on foot and ankle function or dynamic balance in patients.EndNote software was utilized for literature management.RevMan 5.4 software and Cochrane Risk of Bias Assessment Tool were used to evaluate the risk of bias of the included literature.Paired meta-analysis and network meta-analysis of the outcomes such as the Foot and Ankle Ability Measure in daily living subscale score,Foot and Ankle Ability Measure in sports activities subscale score,Star Excursion Balance Test-Anterior score,Star Excursion Balance Test-Posteromedial score,Star Excursion Balance Test-Posterolateral score and Cumberland ankle instability tool score were performed using the network commands of Stata 14.0 software.The strength of evidence rating of the outcome metrics was evaluated according to the GRADE Level of Evidence and Strength of Recommendation Grading Criteria. RESULTS:Of the 22 randomized controlled trials that met the inclusion criteria,1 study was rated as low risk,8 studies were rated as medium risk,and 13 studies were rated as high risk,enrolling a total of 952 patients and 25 treatments.(1)Network meta-analysis showed that compared with the control group,Isokinetic Strength Training,Balance Training,Balance+Stroboscopic Glasses Training,Strength Training,Joint Mobilizations Training,CrossFit Training,CrossFit Training+Self-Mobilization,Wobble Board Training,National Academy of Sport Medicine corrective exercise program,Trigger Point Dry Needling,and Neuromuscular Training had different significant enhancement effects on improving foot and ankle function and dynamic balance in patients with chronic ankle instability(P<0.05).(2)Cumulative probability ranking results showed that the three treatments with the highest ranked Cumberland ankle instability tool score were Joint Mobilizations Training(88.6%)>Visual Feedback Balance Training(83.1%)>CrossFit Training+Self-Mobilization(74.8%);the three treatments with the highest ranked Star Excursion Balance Test-Anterior score were Joint Mobilizations Training(88.4%)>Isokinetic Strength Training(86.9%)>National Academy of Sport Medicine corrective exercise program(65.0%);the three treatments with the highest ranked Star Excursion Balance Test-Posteromedial score were Balance+Stroboscopic Glasses Training(87.4%)>Neuromuscular Training(74.6%)>Strength Training(68.9%);the three treatments with the highest ranked Star Excursion Balance Test-Posterolateral score were CrossFit Training+Self-Mobilization(74.6%)>Balance+Stroboscopic Glasses Training(70.0%)>Neuromuscular Training(63.7%);the three treatments with the highest ranked Foot and Ankle Ability Measure in daily living subscale score were National Academy of Sport Medicine corrective exercise program(91.9%)>Balance+Stroboscopic Glasses Training(85.6%)>Wobble Board Training(82.2%);the three treatments with the highest ranked Foot and Ankle Ability Measure in sports activities subscale score were Balance+Stroboscopic Glasses Training(93.5%)>Balance Training(86.7%)>National Academy of Sport Medicine corrective exercise program(86.4%). CONCLUSION:Non-surgical therapies can significantly improve foot and ankle function and dynamic balance in patients with chronic ankle instability.National Academy of Sport Medicine corrective exercise program had the best efficacy in improving foot and ankle daily activity function in chronic ankle instability patients;Balance+Stroboscopic Glasses Training had the best efficacy in improving foot and ankle sports function and posterior medial dynamic balance;Joint Mobilizations Training had the best efficacy in improving anterolateral dynamic balance and ankle instability condition;and CrossFit Training+Self-Mobilization had the best efficacy in improving posterior lateral dynamic balance.The strength of evidence for each outcome was low,influenced by the risk of methodological bias and risk of publication bias of the included studies.Therefore,the above conclusions need to be validated by more high-quality pilot studies.

Objective To explore the effect of glioma stem cells with high expression of protein tyrosin phosphatase receptor type Z1 (PTPRZ1 )on the phenotypic polarization and phagocytosis of tumor-associated macrophages and its regulatory mechanism.Methods GSCs and non-stem tumor cells (NSTCs) were screened out from human glioblastoma (GBM) specimens using flow cytometry,and the PTPRZ1 expression in paired GSCs and NSTCs were detected.Human peripheral blood mononuclear cells (PBMC)-derived CD14+monocytes were exposed to the conditioned medium from glioma cells or recombinant chemokine C-C motif ligand 20 (CCL20)for TAM polarization.Stable PTPRZ1 knockout GSCs (PTPRZ1-KO GSCs) were constructed using CRISPR/Cas9. TAM phagocytosis to GSCs,NSTCs,PTPRZ1-Control GSCs (PTPRZ1-Ctrl GSCs)and PTPRZ1-KO GSCs and the expression of immunosuppressive phenotype (M2) polarization marker CD163 were examined using flow cytometry.Differentially expressed genes (DEGs ) between paired GSCs and NSTCs were determined using a bulk RNA-sequencing dataset (GSE54791 )from Gene Expression Omnibus (GEO).A gene set informing worse outcome of patients with GBM was generated using The Cancer Genome Atlas (TCGA)-GBM cohort.By intersecting the aforementioned gene set with the gene set that encodes for human membrance proteins,the PTPRZ1 gene is obtained.Gene set enrichment analysis (GSEA)was used for pathway enrichment analysis to compare the differentially regulated pathways between GBMs with high or low PTPRZ1 expression.Bulk RNA sequencing,qRT-PCR and Western blotting were used to identify the DEGs between PTPRZ1-KO GSCs and PTPRZ1-Ctrl GSCs.Results GSCs were more capable of escaping from TAM phagocytosis than NSTCs (P<0.05 )and had specifically up-regulated PTPRZ1 expression.PTPRZ1-KO significantly suppressed GSCs escaping from TAM phagocytosis (P<0.01 ). GBMs with high PTPRZ1 expression showed significant inhibition of pathways mediating phagocytosis (P<0.05).The expression of CCL20 as a M2 TAM polarization chemokine was significantly down-regulated in PTPRZ1-KO GSCs (P<0.05 ).Treatment with recombinant CCL20 up-regulated the expression of CD163 as a M2 TAM marker in TAM.Conclusion PTPRZ1+GSCs mediate M2 TAM polarization and inhibit TAM phagocytosis,which may be related to the up-regulation of CCL20 in PTPRZ1+GSCs.

Objective:To investigate the feasibility of sentinel lymph node biopsy in breast cancer patients with positive axillary lymph nodes turned to clinical negative after neoadjuvant chemotherapy.Methods:Full-text journal databases such as PubMed, Cochrane Library, Embase, Wanfang, VIP, and CNKI were searched to include research literature on sentinel lymph node biopsy in breast cancer patients who had axillary lymph nodes turned negative after neoadjuvant chemotherapy. The retrieval time was self-established to November 2020. Meta-analysis was performed on the literature that met the inclusion criteria. Heterogeneity among studies was analyzed by I2 test. If I2<30%, the heterogeneity among studies was considered to be small. If the value of I2 was between 30% and 70%, it was considered that there was a certain heterogeneity among the studies. If I2> 70%, it was considered that there was great heterogeneity among the studies. Small heterogeneity was analyzed by fixed effects model, otherwise, random effects model was used. Publication bias was evaluated by funnel plot and Egger′s test. Results:Finally, 14 literatures were included, including 4 Chinese literatures and 10 English literatures. The results of Meta-analysis showed that the sentinel lymph node detection rate was 90.7% and the false negative rate was 12.2%.Conclusions:In breast cancer patients with axillary lymph node turning negative, the detection rate of sentinel lymph node biopsy can meet the acceptable clinical standard for sentinel lymph node biopsy, but the false negative rate is still higher than the clinically acceptable standard. It is necessary to screen suitable patients and apply new techniques to reduce the false negative rate of sentinel lymph node biopsy.

Near-infrared fluorescence imaging (NIRFI) is a new noninvasive detection and diagnosis technology, with the continuous development of NIRFI technology, now widely used in the clinic, characterized by high sensitivity, high penetration, no harmful radiation and simple equipment operation. This article describes the recent applications of NIRFI in the diagnosis and treatment of breast cancer and looks at future developments and perspectives in this field.

Objective:To analyze the failure rate of enteral nutrition feeding in patients with mechanical ventilation, explore the influencing factors, and find out the existing problems and deficiencies in the process of nutrition support.Methods:Patients who were admitted to the ICU of Zhejiang People′s Hospital from June 2020 to January 2022 for mechanical ventilation and continuous enteral nutrition through the nose were retrospectively collected as the research subjects. The patients were divided into a standard group and a substandard group. Comparing the feeding status of enteral nutrition patients in the two groups on the 7th day, Logistic regression was used to analyze the influencing factors of feeding substandard.Results:A total of 82 patients (57.34%, 82/143) had feeding failure. Logistic regression analysis showed that complications ( OR=34.47, 95% CI 9.49 to 125.21) and infusion speed ( OR=0.21, 95% CI 0.08 to 0.57) were the influencing factors of feeding failure in ICU mechanical ventilation patients ( P<0.05). Conclusions:The feeding failure rate of ICU mechanical ventilation patients is higher than the feeding failure rate, and the slow infusion speed caused by complications and feeding intolerance is the main reason for the failure. It is suggested to set personalized feeding program for patients, optimize feeding measures, and improve the feeding failure rate.

Objective:To investigate the expression of human epidermal growth factor receptor 2(HER2)and P53 and their relationship with microsatellite instability(MSI)in gastric cancer tissues.Methods:A total of 103 patients diagnosed with gastric cancer between January 2018 and October 2020 at Yueyang Hospital were enrolled in this study.HER2, P53 and mismatch repair proteins in gastric cancer tissues were detected with immunohistochemical(IHC)methods, and MSI screening was conducted at 7 sites with a new Idylla MSI(multiple fluorescent PCR)method.Results:Of 103 gastric cancer patients in this study, 77(74.8%)showed microsatellite stability(MSS)and 26(25.2%)showed MIS via IHC, and PCR also detected 77 MSS cases and 26 MSI cases.In MSI, there was more low HER2 expression than high HER2 expression, and the rate of low HER2 expression in MSI was higher than the rate of high HER2 expression in MSI( P<0.05).Also in MSI, there was more low P53 expression than high P53 expression, and the rate of low P53 expression in MSI was higher than the rate of high expression in MSI( P<0.05). Conclusions:MSS may exist in the process of gastric carcinogenesis and in gastric cancer it may be accompanied by low expression of HER2 and p53 in cancer tissues.There may be a mutually exclusive relationship between MSI and expressions of HER2 and p53, suggesting that carcinogenic mechanisms involving MSI may be very different from those involving HER2 and p53.MSI detection is very valuable in guiding treatment drug selection and prognosis assessment.

Objective:To establish Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines using CRISPR/Cas9 technology and 3D retinal organoid culture.Methods:The target site sequence of H9 cell line was verified by polymerase chain reaction (PCR). SgRNAs were designed by CRISPR/Cas9 technique and their activity was detected.The most optimal sgRNA was selected according to the factors such as activity and specificity.After identification of the target vectors by restriction enzyme and sequencing, the target vectors were transferred to the H9 cell line by electroporation.P2A-tdTomato-P2A-iCreERT2 was inserted between Exon4 and 3’-untranslated region of hES-ZLM-001 gene.Knockin positive clones were obtained after drug treatment, enrichment of positive clones.Primers were designed to perform PCR on the target region, and homozygous de-resistant knockin positive cell clones were selected according to the sequencing results and peaks.The 1-A07 cell line was cultured, and then flow cytometry for the proportion of OCT4 positive cells, immunofluorescence for three stem cell molecular markers including SOX2, NANOG, SSEA4, karyotype analysis were carried out to confirm whether the 1-A07 cell line could be used for further experiments.Retinal organoids were obtained by three-dimensional (3D) culture technology and the expression of molecular markers was detected by immunofluorescence at different developmental stages of retinal organoids. Results:The target site sequence of H9 cell line was consistent with that given by Genebank and Ensembl.Sixteen sgRNAs were designed according to the target site sequence of H9 cell line, and finally sgRNA8 and sgRNA12 were selected.The sgRNAs and recombinant plasmids were transfected into the H9 cell line by electroporation, and four homozygous de-resistant knockin positive cell clones were obtained by PCR.Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines were successfully obtained.In 1-A07 cell line, the proportion of OCT4 positive cells was about 98.7% by flow cytometry, and the expression of three stem cell markers was positive by immunofluorescence, and the karyotype was normal 46, XX.The results showed that the 1-A07 cell line could be used for further experiments.The Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines were differentiated into tdTomato positive retinal organoids by 3D culture technology.BRN3A positive ganglion cells, CALBINDIN positive horizontal cells and CHAT positive amacrine cells appeared on day 30 of differentiation.RECOVERIN positive photoreceptors arose on day 45 of differentiation.PKCα positive bipolar cells presented on day 90 of differentiation.Ganglion cells were shown in the deep layer of retinal organoids, and horizontal cells, amacrine cells and bipolar cells in the middle layer, and photoreceptors in the top layer.Conclusions:Crx-iCreERT2 fluorescent reporter human embryonic stem cell lines are successfully established and can be differentiated into retinal organoids that express tdTomato red fluorescence through 3D culture technology.Those retinal organoids contain the same types of neurons as normal human retinas, and follow a certain temporal and spatial developmental sequence similar to the developmental rules of normal human retinas.Crx-iCreERT2 fluorescent reporter human embryonic stem cell line is a powerful tool for researching retinal development and diseases and can be applied in treatments for blindness.

Whether and how garlic-derived -allylmercaptocysteine (SAMC) inhibits hepatocellular carcinoma (HCC) is largely unknown. In the current study, the role of low-density lipoprotein receptor (LDLR)-related protein 6 (LRP6) in HCC progression and the anti-HCC mechanism of SAMC was examined in clinical sample, cell model and xenograft/orthotopic mouse models. We demonstrated that SAMC inhibited cell proliferation and tumorigenesis, while induced apoptosis of human HCC cells without influencing normal hepatocytes. SAMC directly interacted with Wnt-pathway co-receptor LRP6 on the cell membrane. LRP6 was frequently over-expressed in the tumor tissue of human HCC patients (66.7% of 48 patients) and its over-expression only correlated with the over-expression of -catenin, but not with age, gender, tumor size, stage and metastasis. Deficiency or over-expression of LRP6 in hepatoma cells could partly mimic or counteract the anti-tumor properties of SAMC, respectively. administration of SAMC significantly suppressed the growth of Huh-7 xenograft/orthotopic HCC tumor without causing undesirable side effects. In addition, stable down-regulation of LRP6 in Huh-7 facilitated the anti-HCC effects of SAMC. In conclusion, LRP6 can be a potential therapeutic target of HCC. SAMC is a promising specific anti-tumor agent for treating HCC subtypes with Wnt activation at the hepatoma cell surface.

Objective To investigate the epidemiological features of out-of-hospital patients with ventricular fibrillation (VF) in Shanghai and to analysis factors associated with outcomes, and to provide evidence for improving the success rate of VF.Methods The data of patients with VF admitted to Shanghai Medical Emergency Center from January 2013 to December 2016 were analyzed retrospectively. All the data were recorded including the clinical data, medical service time, return of spontaneous circulation (ROSC) at scene/en route, survival to hospital discharge. Factors that associated with successful resuscitation were analyzed by Logistic regression.Results From 2013 to 2016, 21096 patients with suspected cardiac arrest were admitted to the Shanghai Medical Emergency Center. After excluding ventricular tachycardia (13 cases) and ventricular asystole (20995 cases), 88 patients with VF were enrolled, with 62 male and 26 female; the average age was (63.22±16.15) years old. While bystander cardiopulmonary resuscitation (CPR) was performed in only 21 cases (23.86％). Fifty-seven cases occurred during the day (08:00-20:00), while 31 cases occurred in the night. And the average emergency response time was (6.47±4.13) minutes; the average on-site time was (14.76±10.88) minutes; the average transport to hospital time was (5.95±4.00) minutes. There were no significant differences in response time, on-site time and transport to hospital time each year, and there were no significant differences in emergency medical service time between day and night either. From 2013 to 2016, prehospital successful resuscitation rate was decreased by years [95.65％ (22/23), 87.50％ (14/16), 83.33％ (20/24) vs. 80.00％(20/25), respectively,χ2 = 1.895,P = 0.595]. Survival to hospital discharge rate was increased by years [21.74％ (5/23), 31.25％ (5/16), 37.50％ (9/24), 40.00％ (10/25), respectively,χ2 = 2.862,P = 0.413]. The success rate of prehospital resuscitation for patients with 1, 2, ≥3 defibrillation was 35.23％ (31/88), 23.08％ (12/52), 89.19％ (33/37), respectively (χ2 = 42.811,P = 0.000). The on-site time in successful final resuscitation group was shorter than that in final resuscitation failure group (minutes: 10.85±8.83 vs. 16.79±11.36,t = 2.367,P = 0.020), the ROSC time in successful final resuscitation group was shorter than that of final resuscitation failure group (minutes: 3.24±3.17 vs. 7.43±6.64, t = 3.175,P = 0.002). It was shown by Logistic regression that long ROSC time was the risk factor for final resuscitation failure [odds ratio (OR) = 0.771,P = 0.024]. Gender, age, availability of witnesses CPR, call time, emergency response time, on-site time and transport to hospital time had no significant impact on the prehospital successful resuscitation and final successful resuscitation. In prehospital successful resuscitation group, there was significant difference in survival to hospital discharge rate among different defibrillation times group [48.39％ (15/31), 58.33％ (7/12) vs. 21.21％ (7/33),χ2 = 7.460,P = 0.024].Conclusions From 2013 to 2016, there were no significant changes in the emergency response time, prehospital successful resuscitation rate and survival to hospital discharge rate of patients with VF in Shanghai. Though, repeated defibrillation could significantly increased prehospital successful resuscitation rate, multiple defibrillation indicated decline of survival to hospital discharge rate in prehospital successful resuscitation group. Additionally, long on-site time and long ROSC time indicated poor prognosis.

Objective To study the curative effect of XRCC2 gene silencing mediated by shRNA combined with radiation on human colonic trans?planted carcinoma in nude mice. Methods Colonic carcinoma T84 cells were transfered into BALB/c nude mice to establish a tumor xenograft mod?el in vivo. Mice were divided into three groups:control,shRNA?SC and shRNA?XRCC2 and exposed to X?ray radiation. The change of volume and weight of the xenografts were examined after receiving radiotherapy and the pathological analysis of tumor tissues were conducted. Results Tumor xenografts transfected with shRNA?XRCC2 in nude mice grew slowly. The xenograft volume in the shRNA?XRCC2 group was decreased significant?ly from day 12 to day 28 after radiotherapy compared with the control group(P<0.01). The xenograft weight in the shRNA?XRCC2 group was small?er than in the control group,with statistically significant difference(t=18.843,P<0.01). The inhibited rate of xenografts in the shRNA?XRCC2 group(56.25%),was markedly higher than that in the shRNA?SC group(4.69%). Pathological analysis of colonic transplanted carcinoma showed that nuclear atypia was not obvious,karyokinesis was decreased and small areas of necrosis were present in tumor xenografts treated with shRNA?XRCC2 transfection. Conclusion XRCC2 gene silencing combined with radiation has significant inhibition effect on colonic transplanted carcino?ma in nude mice.