BACKGROUND:Studies demonstrated that miR-135a-5p was highly expressed in mouse embryonic palatal mesenchymal cells with cleft palate induced by dexamethasone.The primary cilium and its mediated Shh signaling pathway were involved in the autophagy of mouse embryonic palatal mesenchymal cells.It is speculated that miR-135a-5p may regulate autophagy in mouse embryonic palatal mesenchymal cells through primary cilia and its mediated Shh signaling pathway. OBJECTIVE:To investigate the regulatory effect of miR-135a-5p on autophagy of mouse embryonic palatal mesenchymal cells. METHODS:In vitro,palatal mesenchymal cells from C57BL/6J mouse embryos were extracted and cultured.Cell transfections were set up as follows:(1)the cells were divided into control group,miR-135a-5p negative control group and miR-135a-5p mimic group;(2)NC+miR-NC group,KIF3B overexpression group,and miR-135a-5p+KIF3B group:qRT-PCR was performed to verify transfection efficiency of miR-135a-5p and KIF3B.A transmission electron microscope was used to observe the number of autophagosome/autophagolysosome in the cells of each group.The degree of fluorescence expression of autophagy marker LC3B was determined by the immunofluorescence technique.The protein expression of KIF3B,LC3 and P62 was determined by western blot assay.(3)The cells were divided into miR-135a-5p negative control group,and SAG treated group,and SAG+miR-135a-5p group.qRT-PCR was used to detect the mRNA expression levels of Gli3,a key transcription factor downstream of Shh signaling.The protein expressions of autophagy-related proteins LC3 and P62 were detected by western blot assay. RESULTS AND CONCLUSION:(1)After overexpression of miR-135a-5p,the number of autophagosome/autophagolysosome was significantly increased(P<0.01).The fluorescence density of LC3B increased significantly(P<0.01);the protein expression of KIF3B and P62 decreased(P<0.01),and the protein expression of LC3 increased.(2)After overexpression of KIF3B,the number of autophagosome/autophagolysosome was significantly decreased(P<0.01);the fluorescence density of LC3B was decreased(P<0.01);the protein expression of P62 was increased(P<0.01),and the protein expression of LC3 was decreased(P<0.01).Targeted expression of KIF3B was inhibited by miR-135a-5p(P<0.01);the number of autophagosome/autophagolysosome,the fluorescence intensity of LC3B as well as the protein expression of LC3 were reversed(P<0.01)and the protein expression of P62 was decreased(P<0.01).(3)SAG significantly increased the mRNA expression of Gli3(P<0.01),increased the protein expression of P62(P<0.01),and decreased the protein expression of LC3(P<0.01).When miR-135a-5p was added,Gli3 mRNA expression was significantly decreased(P<0.01);P62 protein expression was decreased(P<0.01),and LC3 protein expression was reversed(P<0.01).(4)These results indicate that miR-135a-5p targets the inhibition of KIF3B and promotes autophagy in mouse embryonic mesenchymal cells possibly by negatively regulating the Shh signaling pathway.

Objective:To analyze the short-term clinical outcomes of total laparoscopic distal gastrectomy (TLDG) and laparoscopic-assisted distal gastrectomy (LADG) combined with Billroth-Ⅱ+Braun anastomosis.Methods:Clinical characteristics of patients undergoing laparoscopic distal gastrectomy combined with Billroth-Ⅱ+Braun anastomosis at Department of Gastrointestinal Surgery, Shandong Provincial Hospital Affiliated to Shandong First Medical University from Jan 2020 to Oct 2022 were analyzed. Patients were divided into TLDG group ( n=62) and LADG group ( n=62) according to the surgical approach. Results:There were significant differences in the preoperative clinical data section between the two groups, and 124 patients (62 in each group) were enrolled after using propensity score matching to balance significant variables. Compared with the LADG group, the TLDG group showed statistically differences in time to first venting [(2.9±1.3) vs. (2.3±0.8) d, Z=-3.072, P=0.002], time to first fluid diet [(5.9±1.3) vs. (5.4±1.4) d, Z=-2.031, P=0.042] and incision length [(7.1±1.4) vs. (4.8±0.8) cm, Z=-6.331, P=0.000]. Total postoperative complication rate in the TLDG group and the LADG group (29% vs. 37%, χ2=0.911, P=0.340) was not statistically significant. Incidence of postoperative pneumonia was lower in the TLDG group than in the LADG group (3% vs. 13%, χ2=3.916, P=0.048), and incidence of all remaining postoperative complications were not statistically significant. There was no statistically significant difference in the incidence of serious postoperative complications between the TLDG and LADG groups ( P=1.000). Multifactorial analysis revealed that male ( P=0.023) and age ≥65 years ( P=0.001) were independent risk factors for postoperative complications. Conclusion:TLDG is safe and feasible and has better short-term clinical efficacy than LADG.

This study aims to analyze the main drug resistance mechanism of Salmonella enteritidis induced by polymyxin in vitro.In this study,the resistance of Salmonella enteritidis CMCC(B)50335(ZK)to polymyxin was induced in vitro,and the growth characteristics,exercise ability,ul-trastructure and sensitivity to 16 antimicrobial agents before and after induction were determined by turbidimetry,semi-solid agar method,transmission electron microscope and disk diffusion method,and the whole genome and single nucleotide polymorphism(SNP)were detected by Illu-mina NovaSeq PE150 method.RT-qPCR was used to detect the differences in the expression levels of six drug-resistant related genes.The recombinant strain △phoPE1-128-1,which induced drug-resistant strain E1-128-1,was constructed by homologous recombination of Red Escherichia coli,and its sensitivity to polymyxin was detected.The results showed that three strains of induced drug-resistant bacteria E1-128-1,E1-128-3 and E2-128-3 were screened out,and the MIC increased by 128,64 and 64 times respectively after drug resistance stability test.Induced drug resistance had no significant effect on the growth ability of the tested bacteria and the sensitivity of 16 antibacte-rial drugs.The exercise ability of E1-128-1was significantly increased,and the cell wall and plasma membrane obviously thicken.There was no significant difference in the genome components of E1-128-1 before and after induction,but eight missense mutations of six drug-resistance related genes,including phoP/phoQ,cpxP,lptD,csrA and acrB,were detected,including four missense mutation sites of phoP,namely Leu185Trp,His189Ser,Thr190Tyr and Ile191His.The corresponding genes were sequenced by PCR,and the results were consistent with those of SNP.RT-qPCR results showed that the expression levels of mutant genes of the three induced strains increased signifi-cantly.Compared with E1-128-1,the MIC of △phoP E1-128-1 decreased to 1 mg/L.It is sug-gested that the mutation and increased expression of phoP gene are important factors for inducing polymyxin resistance of Salmonella CMCC(B)50335 in vitro.

BACKGROUND: LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer. METHODS: We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels. RESULTS: On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]). CONCLUSION: LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile. TRIAL REGISTRATION ClinicalTrials.gov, NCT04563936.
Humans ; Male ; Antineoplastic Agents, Hormonal/therapeutic use* ; East Asian People ; Gonadotropin-Releasing Hormone/agonists* ; Goserelin/therapeutic use* ; Prostate-Specific Antigen ; Prostatic Neoplasms/drug therapy* ; Testosterone

BACKGROUND: The KangDuo-Surgical Robot-01 (KD-SR-01) system is a new surgical robot recently developed in China. The aim of this study was to present our single-center experience and mid-term outcomes of urological procedures using the KD-SR-01 system. METHODS: From August 2020 to April 2023, consecutive urologic procedures were performed at Peking University First Hospital using the KD-SR-01 system. The clinical features, perioperative data, and follow-up outcomes were prospectively collected and analyzed. RESULTS: A total of 110 consecutive patients were recruited. Among these patients, 28 underwent partial nephrectomy (PN), 41 underwent urinary tract reconstruction (26 underwent pyeloplasty, 3 underwent ureteral reconstruction and 12 underwent ureterovesical reimplantation [UR]), and 41 underwent radical prostatectomy (RP). The median operative time for PN was 112.5 min, 157.0 min for pyeloplasty, 151.0 min for ureteral reconstruction, 142.5 min for UR, and 138.0 min for RP. The median intraoperative blood loss was 10 mL for PN, 10 mL for pyeloplasty, 30 mL for ureteral reconstruction, 20 mL for UR, and 50 mL for RP. All procedures were successfully completed without conversion, and there were no major complications in any patient. The median warm ischemia time of PN was 17.3 min, and positive surgical margin was not noted in any patient. The overall positive surgical margin rate of RP was 39% (16/41), and no biochemical recurrence was observed in any RP patient during the median follow-up of 11.0 months. The surgical success rates of pyeloplasty and UR were 96% (25/26) and 92% (11/12) during the median follow-up of 29.5 months and 11.5 months, respectively. CONCLUSION The KD-SR-01 system appears feasible, safe, and effective for most urological procedures, based on our single-center experience.
Male ; Humans ; Robotic Surgical Procedures/methods* ; Robotics ; Treatment Outcome ; Retrospective Studies ; Ureter/surgery* ; Urologic Surgical Procedures/methods* ; Laparoscopy/methods*

Objective:To develop and validate a nomogram model for predicting microvascular invasion (MVI) in hepatocellular carcinoma (HCC) based on preoperative enhanced computed tomography imaging features and clinical data.Methods:The clinical data of 210 patients with HCC undergoing surgery in the Second Affiliated Hospital of Anhui Medical University from May 2018 to May 2022 were retrospectively analyzed, including 172 males and 38 females, aged (59±10) years old. Patients were randomly divided into the training group ( n=147) and validation group ( n=63) by systematic sampling at a ratio of 7∶3. Preoperative enhanced computed tomography imaging features and clinical data of the patients were collected. Logistic regression was conducted to analyze the risk factors for HCC with MVI, and a nomogram model containing the risk factors was established and validated. The diagnostic efficacy of predicting MVI status in patients with HCC was assessed by receiver operating characteristic (ROC) curve, calibration curves, decision curve analysis (DCA), and clinical impact curve (CIC) of the subjects in the training and validation groups. Results:The results of multifactorial analysis showed that alpha fetoprotein ≥400 μg/ml, intra-tumor necrosis, tumor length diameter ≥3 cm, unclear tumor border, and subfoci around the tumor were independent risk factors predicting MVI in HCC. A nomogram model was established based on the above factors, in which the area under the curve (AUC) of ROC were 0.866 (95% CI: 0.807-0.924) and 0.834 (95% CI: 0.729-0.939) in the training and validation groups, respectively. The DCA results showed that the predictive model thresholds when the net return is >0 ranging from 7% to 93% and 12% to 87% in the training and validation groups, respectively. The CIC results showed that the group of patients with predictive MVI by the nomogram model are highly matched with the group of patients with confirmed MVI. Conclusion:The nomogram model based on the imaging features and clinical data could predict the MVI in HCC patients prior to surgery.

Objective:To evaluate the safety and effectiveness of Kangduo endoscopic surgical robot system for partial nephrectomy.Methods:Consecutive patients with stage T 1 renal tumor meeting the inclusion criteria from the Department of Urology, Peking University First Hospital from December 2020 to February 2021 were prospectively enrolled. All patients underwent partial nephrectomy with the Kangduo endoscopic surgical robot system after signing the informed consent. Clinical data including preoperative, perioperative and postoperative pathology and follow-up were collected. Results:Among the 26 patients, there were 16 males and 10 females, with a median age of 53(33-74) years, and a median body mass index of 25.99(20.90-32.91) kg/m 2. There were 12 cases of left kidney tumor and 14 cases of right kidney tumor. The median tumor diameter was 2.2(1.0-3.5) cm. The median time of warm ischemia was 17.7(7.1-29.2) minutes, and all of them were less than 30 minutes. The median docking time was 4.7(2.3-9.9) minutes, and the median time of robotic arm operation was 65.0 (37.0-155.0) minutes. The median National Aeronautics and Space Administration Task Load Index (NASA-TLX) score was 5.3 (2.0-28.0), and no instrument-related adverse events occurred intraoperatively. The median postoperative hospital stay was 4 (4-5) days. All tumor margins were negative on pathologic reports. No Clavien Ⅱ stage operative complications occurred in all patients during perioperative period and 1 month after the surgery. Conclusions:The partial nephrectomy using the kangduo endoscopic surgical robot system were completed successfully, and no instrument-related adverse events and complications occurred, showing that this surgical system used for partial nephrectomy is safe and effective.

Objective:To investigate the effect of polyethylene glycol combined with simethicone on boston bowel preparation scale(BBPS) score and tolerance in patients with colonoscopy.Methods:From January 2018 to January 2019, 220 patients who underwent colonoscopy in Beijing Shijitan Hospital Affiliated to Capital Medical University were selected as the research objects.According to different bowel preparation schemes, they were divided into control group and observation group, 110 cases in each group.In the control group, 4 bags of compound polyethylene glycol electrolyte powder were added with cold boiled water to prepare 4 L solution for intestinal preparation.Starting 6 hours before endoscopy, 1 L solution was taken every hour, and the interval from the last time to the beginning of colonoscopy was≤4 h. On the basis of the control group, the observation group was given 30 mL simethicone emulsion 4 hours before endoscopy on the basis of the control group.The BBPS score, intracavitary liquid volume score, total score, tolerance and adverse reactions were compared between the two groups.Results:The whole colon score (7.16±0.61), left colon score (2.89±0.62), transverse colon score (2.78±0.64) and right colon score (1.58±0.49), intestinal cavity fluid score (1.47±0.48) and total score (8.84±0.87) of the observation group were higher than those of the control group ((5.13±0.76), (2.23±0.86), (2.15±0.76), (1.14±0.16), (0.91±0.55), (7.11±1.04)), and the differences were statistically significant ( t=21.854, 6.532, 6.652, 8.957, 8.054, 13.380, all P<0.05). The tolerance of the observation group (90.00%(99/110)) was higher than that of the control group (81.81%(90/110)), but the difference was not statistically significant( χ 2=3.043, P=0.081). The incidence of abdominal distension in the observation group (1.82%(2/110)) was lower than that in the control group (8.18%(9/110)), the difference was statistically significant(χ 2=4.690, P<0.05), and there was no significant difference in the incidence of electrolyte disorder, nausea and vomiting, hypoglycemia or hunger, palpitation and chest tightness between the two groups (all P>0.05). Conclusion:The application of polyethylene glycol combined with Spanish silicone oil in intestinal preparation can improve the intestinal cleanliness of patients, but does not increase the tolerance of patients compared with polyethylene glycol alone, but significantly reduces the incidence of abdominal distension.

Objective:To investigate the prevalence, gene variation and prognosis of very long chain acyl CoA dehydrogenase deficiency (VLCADD) in newborns in Henan Province.Methods:From January 2013 to December 2019, 867 103 newborns were investigated for VLCADD by tandem mass spectrometry.Children who diagnosed as VLCADD and their families were subjected to next-generation sequencing and Sanger sequencing.Clinical data, biochemical changes and gene variation characteristics of the confirmed cases of VLCADD were analyzed.Dietary guidance was given, and their growth and development were followed up.Results:Six neonates were diagnosed as VLCADD, and the prevalence of VLCADD in the Henan Province was 1/144 517.A total of 11 mutations in the ACADVL gene were found, including 5 new variants c. 692-2_692-1delAG, c.753-23_753-22del, c.960delG, c.1361A>G, and c. 1955C>T.The newborns were given a high-carbohydrate, low-fat diet, and followed up for 8-56 months.Except for two deaths, all patients had a good outcome. Conclusions:The prevalence of neonatal VLCADD in Henan Province is 1/144 517.This results has enriched the ACADVL gene mutation spectrum and provided an important basis for the screening and diagnosis of VLCADD.

Objective: To identify the initial chest computed tomography (CT) findings and clinical characteristics associated with the course of coronavirus disease 2019 (COVID-19) pneumonia. Materials and Methods: Baseline CT scans and clinical and laboratory data of 72 patients admitted with COVID-19 pneumonia (39 men, 46.2 ± 15.9 years) were retrospectively analyzed. Baseline CT findings including lobar distribution, presence of ground glass opacities, consolidation, linear opacities, and lung severity score were evaluated. The outcome event was recovery with hospital discharge. The time from symptom onset to discharge or the end of follow-up (for those remained hospitalized) was recorded. Data were censored in events such as death or discharge without recovery. Multivariable Cox proportional hazard regression was used to explore the association between initial CT, clinical or laboratory findings, and discharge with recovery, whereby hazard ratio (HR) values < 1 indicated a lower rate of discharge at four weeks and longer time until discharge. Results: Thirty-two patients recovered and were discharged during the study period with a median length of admission of 16 days (range, 9 to 25 days), while the rest remained hospitalized at the end of this study (median, 17.5 days; range, 4 to 27 days). None died during the study period. After controlling for age, onset time, lesion characteristics, number of lung lobes affected, and bilateral involvement, the lung severity score on baseline CT (> 4 vs. ≤ 4 [reference]: adjusted HR = 0.41 [95% confidence interval, CI = 0.18–0.92], p = 0.031) and initial lymphocyte count (reduced vs. normal or elevated [reference]: adjusted HR = 0.14 [95% CI = 0.03–0.60], p = 0.008) were two significant independent factors that influenced recovery and discharge. Conclusion Lung severity score > 4 and reduced lymphocyte count at initial evaluation were independently associated with a significantly lower rate of recovery and discharge and extended hospitalization in patients admitted for COVID-19 pneumonia.