Objective To investigate the effects of ROCK-siRNA transfection on endothelial cell senescence and endothelial microparticles (EMPs) induced by angiotensin II (Ang II). Methods Human umbilical vein endothelial cells (HUVECs) were treated with Ang II (1.0 μmo/L) to induce cellular senescence models, followed by transfection with ROCK-siRNA. The cells were divided into four groups: control group, model group, negative transfection control group (Ang II combined with NC-siRNA), and ROCK-siRNA transfection group (Ang II combined with ROCK-siRNA). Cellular senescence was assessed by SA-β-Gal staining. EMP levels in cell supernatants and intracellular reactive oxygen species (ROS) levels were assessed using flow cytometry. The expression levels of silenced information regulator 1(SIRT1) and p53 protein in each group were analyzed by Western blotting. Results Following ROCK-siRNA transfection, the number of senescent cells induced by Ang II was significantly reduced, accompanied by decreased CD31⁺ EMP levels and suppressed intracellular ROS levels. Meanwhile, the expression levels of SIRT1 were up-regulated, while the expression levels of p53 were down-regulated. Conclusion Silencing ROCK expression suppresses EMP release, reduces ROS generation, regulates the expression of SIRT1 and p53, and ultimately attenuates Ang II-induced endothelial cell senescence.
Humans ; Angiotensin II/pharmacology* ; Cellular Senescence/genetics* ; Human Umbilical Vein Endothelial Cells/cytology* ; RNA, Small Interfering/metabolism* ; Reactive Oxygen Species/metabolism* ; Sirtuin 1/genetics* ; Transfection ; Tumor Suppressor Protein p53/genetics* ; Cell-Derived Microparticles/drug effects* ; rho-Associated Kinases/metabolism* ; Endothelial Cells/metabolism* ; Cells, Cultured

Objective To analyze the epidemiological characteristics and trends of the incidence and mortality of biliary tract cancer in Shanghai from 2002 to 2020. Methods Data on new cases and deaths of malignant tumors of the gallbladder, extrahepatic bile ducts, and other biliary tract organs from 2002 to 2020 were obtained from the Population-based Cancer Registry and Vital Statistics System of Shanghai Municipal Center for Disease Control and Prevention. Cases or deaths, proportion, crude rate, age-specific rate, age-standardized rate (ASR) and others were calculated stratified by year of diagnosis or death, gender and age-group. ASRs were calculated using Segi′s 1960 world standard population. Trends of the annual percent change (APC) of ASRs, age-specific rates and proportions of new cases with selected diagnostic character of biliary tract cancer stratified by different groups were analyzed by Joinpoint analysis software. Results Annual new cases of biliary tract cancer in Shanghai increased from 963 in 2002 to 1 537 in 2020, with ASR of incidence changing from 3.91/10⁵ to 3.59/10⁵. Annual deaths increased from 830 to 1 225, with ASR of mortality decreased from 3.36/10⁵ to 2.69/10⁵. In 2020, the crude rate of incidence of biliary tract cancer was 10.43/10⁵ (9.54/10⁵ in males and 11.30/10⁵ in females) in Shanghai, and the ASR was 3.59/10⁵ (3.54/10⁵ in males and 3.61/10⁵ in females), with no statistically significant gender difference (P=0.731). The crude rate of mortality was 8.31/10⁵ (7.60/10⁵ in males and 9.00/10⁵ in females), and the ASR was 2.69/10⁵ (2.69/10⁵ in males and 2.66/10⁵ in females), also with no significant gender difference (P=0.874). Age-specific nunbers and rates of incidence and mortality generally increased with aging. Stratified by gender, the trend of ASRs of incidence of biliary tract cancer in Shanghai in males showed no significant change (P=0.179) from 2002 to 2020, nor did that of ASRs of mortality (P=0.738). In females, the ASRs of incidence decreased at an average annual rate of 1.58% (P<0.001), while the trend of ASRs of mortality showed no significant change from 2002 to 2011 (P=0.774), but ASRs decreased at an average annual rate of 3.72% from 2011 to 2020 (P<0.001). Among new cases, the proportions of morphological verification increased, while the proportions of imaging verification decreased. The gallbladder was the most common site, but its proportions decreased significantly, whereas the proportions of extrahepatic bile duct increased from 25.75% to 42.88%. Over 60% of cases were unknown stage at diagnosis, while the combined proportions of stages Ⅰ-Ⅲ remained less than that of stage Ⅳ. Conclusions The ASRs for incidence and mortality of biliary tract cancer in Shanghai remain relatively high, with distinct epidemiological characteristics. The improvement in the diagnosis and treatment of biliary diseases maybe have impact on the incidence patterns of biliary tract cancer in Shanghai, but the effect on increasing survival rates and reducing mortality rates is relatively lagging. It needs a big progress to advance the screening, diagnosis, and survival of biliary tract cancer in Shanghai. This study provides a foundation for further research and prevention strategies for biliary tract cancer.

A pair of positively charged stable isotope labelling agents,4-((2,5-dioxocyclopentyloxy)carbonyl)-N,N,N-trimethylnaphthalen-1-aminium iodide(DPTNA)and its deuterated counterpart d3-DPTNA,were designed and synthesized,which were applied to pre-column labelling of alkylphenols to enhance the sensitivity and accuracy of liquid chromatography-tandem mass spectrometry(LC-MS/MS).By crosslinking covalent organic framework(COF)with hydrophilic sodium alginate and Ca2+in the presence of pore forming agent NH4HCO3 and surfactant P123,abundant of"gas pocket"were created before and during the beads formation process,and high porous COF beads with macroporous and open structure were achieved after removal of gas.Compared with traditional powdered adsorbents,COF beads demonstrated superior operability and recyclability.The proposed method was applied to analysis of alkylphenols in water samples with method LODs of 0.25-0.50 ng/L,while the method LOQs were in the range of 0.80-1.52 ng/L,and recoveries ranged from 92.2%to 98.3%.The proposed method could be well applied to the simultaneous enrichment and analysis of alkylphenols in water samples.

Objective To explore the spatiotemporal expression patterns of Patched 1(Ptch1)and insulin enhancer binding protein-1(Isl 1)in mouse embryonic foregut and their relationship with the early development of trachea-main bronchus.Methods The foregut of 60 mouse embryos at E9.5-12.5 was separated for the detection of Isl1 and sonic hedgehog(Shh)protein by Western blotting.Serial paraffin sections of 6 mouse embryos at E9.5-14.5 were taken for immunohistochemical staining and immunofluorescence double staining with Isl1,Ptch1,forkhead box protein A2(Foxa2),type Ⅱ collagen α1 chain(Col2a1)and α-smooth muscle actin(α-SMA),as well as HE staining and Masson staining.Results The expression trend of Isl1 and Shh in foregut endoderm at E9.5-12.5 was similar,and the peak of Shh expression was later than Isl1.The foregut developed into the trachea at E9.5-12.5,Ptch1 was expressed in the thickening and protrusion of the respiratory endoderm,the laryngal-tracheal groove and the solid cell cord,accompanied by the increase and aggregation of Isl1-positive mesenchymal cells,forming a characteristic pyramidal structure centered on the respiratory endoderm and the solid cell cord;The main bronchus appeared at E12.5-13.5,Ptch1 was only expressed in its lateral wall,accompanied by the accumulation of Isl1-positive mesenchymal cells;The trachea-main bronchial epithelium lost Ptch1 expression and the surrounding Isl 1-positive mesenchymal cells also decreased rapidly at E13.5-14.5.Co12a1-positive chondrocytes first appeared in the Isl1-positive mesenchymal area adjacent to the Ptch1-positive epithelium at E12.5;Col2a1-positive cartilage was nested within the Isl1-positive mesenchymic area in a"C"shape and expanded in a proximal-distal pattern at E12.5-13.5;Col2a1-positive cartilage extended to the dorsal trachea beyond the Isl1-positive mesenchyma and encircles α-SMA positive smooth muscle in a circular manner at E14.5.Conclusion The expression of Ptch1 in the foregut endoderm is involved in the development and morphogenesis of the trachea-main bronchus epithelium,and is closely related to the proliferation and aggregation of Isl1-positive mesenchyme in the trachea-main bronchial wall,Subsequently,they jointly determine the time,location and extent of airway cartilage.

Objective To investigate the in-hospital screening results of developmental dysplasia of the hip (DDH) in infants and young children in Ili area, and to analyze the risk factors affecting the occurrence of DDH. Methods According to the cluster sampling method 5 536 infants and young children who underwent DDH screening in the pediatric outpatient department and orthopedic outpatient department of our hospital from December 2019 to June 2022 were selected as the research objects. The children who met the diagnostic criteria of DDH were selected as the observation group (n=35), and 100 normal children were selected as the control group. Univariate and multivariate analysis were used to determine the independent risk factors affecting the occurrence of DDH in infants. Results Among the 39 cases were positive in primary screening, 35 cases were positive in secondary screening, and the positive rate was 6.32‰ . The results of single factor analysis showed that the proportion of women, second birth and above, caesarean section, breech delivery, family history, high altitude area, living environment room temperature < 18^°C, and leg binding when swaddling in the observation group was higher than that in the control group (P<0.05) . Logistic regression analysis showed that mode of production, region, room temperature of living environment and swaddling mode were independent risk factors for DDH in infants (P<0.05). Conclusion Caesarean section, high altitude area, living environment room temperature < 18^°C and leg binding in infants are related to the occurrence of DDH in infants, which can provide some reference for clinical screening, diagnosis and treatment.

Objective To explore characteristics of co-infection of Chlamydia pneumoniae(Cpn)and severe acute respiratory syndrome coronavirus 2(SARS-CoV-2),and identify their effect on SARS-CoV-2-induced inflammatory response.Methods Patients with coronavirus disease 2019(COVID-19)who received treatment in a hospital in Chenzhou City from December 20,2022 to February 20,2023 were selected.According to the severity of COVID-19,severe and critical cases were classified as the severe symptom group,while mild and moderate cases were classified as the mild symptom group.Meanwhile,according to the age of patients(≥18 years old as adults,＜18 years old as juveniles),they were divided into the adult severe symptom group,adult mild symptom group,juvenile severe symptom group,and juvenile mild symptom group.Propensity score was adopted to match age,gender,and under-lying diseases of patients in severe symptom and mild symptom group in a 1∶1 ratio.Bronchoalveolar lavage fluid(BALF),throat swabs,and serum specimens of patients were collected.Cpn IgG/IgM antibody was detected by enzyme-linked immunosorbent assay(ELISA),levels of 12 common cytokines(including interleukin-8[IL-8])in BALF were detected by flow cytometry,differences among groups were compared.Results A total of 102 patients were included,with 61 severe and critical(severe symptom)patients,as well as 41 mild and moderate(mild symp-tom)patients.There were 71 patients aged ≥18 years and 31 juvenile patients aged＜18 years.There were 39 pa-tients in the adult severe symptom group and 32 in the adult mild symptom group,and 30 pairs were successfully matched through propensity score analysis.There were 22 patients in the juvenile severe symptom group and 9 in the juvenile mild symptom group,and 8 pairs were successfully matched through propensity score analysis.Among COVID-19 patients,the positive rates of Cpn IgG and IgM were 36.27％(n=37)and 8.82％(n=9),respective-ly,with 1 case positive for both Cpn IgG and IgM.The level of interferon(IFN)-α in serum specimens from adult patients with severe symptom combined with positive Cpn IgG was higher than that of IgG negative patients(P=0.037).There was no statistically significant difference in the levels of other cytokines in BALF and serum speci-mens between the two groups of patients(all P＞0.05).The levels of IL-8 and IL-17 in serum specimens of patients with positive Cpn IgG in the adult mild symptom group were both higher than those in Cpn IgG negative patients(both P＜0.05).The levels of IL-8 in both BALF and serum specimens from Cpn IgM positivity patients in the ju-venile mild symptom group were higher than those from patients with negative Cpn IgM(both P＜0.05).Logistic regression analysis results showed that Cpn IgG and IgM positivity were not risk factors for the development of se-vere COVID-19.Conclusion Combined Cpn infection is not a risk factor for the development of severe symptom in COVID-19 patients,and Cpn infection has limited impact on the secretion of inflammatory factors caused by SARS-CoV-2.

Objective To study the anti-reflux effect of conical gastric stump embedding in radical resection of esophageal cancer.Methods A total of 60 patients who planned to undergo radical resection of esophageal cancer in our hospital from June 2020 to June 2022 were selected as the study objects,and the patients were divided into the observation group and the control group by random number table method,with 30 cases in each group.Patients in both groups underwent laparoscopic radical resection of esophageal cancer and esophagogastric end-to-side mechanical anastomosis.The observation group adopted the conical gastric stump embedding technique after esophagogastric end-to-side mechanical anastomosis.The perioperative related indexes,postoperative complications and gastroesophageal reflux of patients in the two groups were compared.The postoperative anti-reflux effect was evaluated by reflux disease questionnaire(RDQ)score and 24-hour intraesophageal pH monitoring.Results The operation time and digestive tract reconstruction time of patients in the observation group were longer than those in the control group(P<0.05),while there was no statistically significant difference in the amount of intraoperative bleeding,the number of lymph node dissection,the first exhaust time,or the postoperative hospital stay of patients between the two groups(P>0.05).There was no statistically significant difference in the overall incidence of postoperative complications between the two groups(P>0.05).The severity of postoperative gastroesophageal reflux of patients in the observation group was lighter than that in the control group,and the difference was statistically significant(P<0.05).The RDQ score,24-hour reflux frequency,>5 minutes reflux frequency,pH<4 time,and longest reflux time of patients in the observation group was significantly lower/less/shorter than those in the control group(P<0.05).Conclusion The conical gastric stump embedding technique is safe and feasible in the radical resection of esophageal cancer.Although the operation time and digestive tract reconstruction time are slightly prolonged,it does not increase the perioperative risks,which can significantly reduce the occurrence and severity of postoperative gastroesophageal reflux of patients,and achieve a good anti-reflux effect.

Objective To monitor the susceptibility of clinical isolates to antimicrobial agents in tertiary hospitals in major regions of China in 2022.Methods Clinical isolates from 58 hospitals in China were tested for antimicrobial susceptibility using a unified protocol based on disc diffusion method or automated testing systems.Results were interpreted using the 2022 Clinical &Laboratory Standards Institute(CLSI)breakpoints.Results A total of 318 013 clinical isolates were collected from January 1,2022 to December 31,2022,of which 29.5％were gram-positive and 70.5％were gram-negative.The prevalence of methicillin-resistant strains in Staphylococcus aureus,Staphylococcus epidermidis and other coagulase-negative Staphylococcus species(excluding Staphylococcus pseudintermedius and Staphylococcus schleiferi)was 28.3％,76.7％and 77.9％,respectively.Overall,94.0％of MRSA strains were susceptible to trimethoprim-sulfamethoxazole and 90.8％of MRSE strains were susceptible to rifampicin.No vancomycin-resistant strains were found.Enterococcus faecalis showed significantly lower resistance rates to most antimicrobial agents tested than Enterococcus faecium.A few vancomycin-resistant strains were identified in both E.faecalis and E.faecium.The prevalence of penicillin-susceptible Streptococcus pneumoniae was 94.2％in the isolates from children and 95.7％in the isolates from adults.The resistance rate to carbapenems was lower than 13.1％in most Enterobacterales species except for Klebsiella,21.7％-23.1％of which were resistant to carbapenems.Most Enterobacterales isolates were highly susceptible to tigecycline,colistin and polymyxin B,with resistance rates ranging from 0.1％to 13.3％.The prevalence of meropenem-resistant strains decreased from 23.5％in 2019 to 18.0％in 2022 in Pseudomonas aeruginosa,and decreased from 79.0％in 2019 to 72.5％in 2022 in Acinetobacter baumannii.Conclusions The resistance of clinical isolates to the commonly used antimicrobial agents is still increasing in tertiary hospitals.However,the prevalence of important carbapenem-resistant organisms such as carbapenem-resistant K.pneumoniae,P.aeruginosa,and A.baumannii showed a downward trend in recent years.This finding suggests that the strategy of combining antimicrobial resistance surveillance with multidisciplinary concerted action works well in curbing the spread of resistant bacteria.

Objective:To investigate the influences of Neferine(Nef)on inflammatory injury in nephrotic syndrome(NS)rats by regulating the MAPK/NF-κB pathway.Methods:SD rats were separated into control check group(CK group),Model group,low-dose Nef group(Nef-L group,2.5 mg/kg),high-dose Nef group(Nef-H group,5 mg/kg),prednisone acetate group(PA group,6.3 mg/kg),Anisomycin(MAPK agonist)group(5 μmol/L),Nef-H+Anisomycin group(5 mg/kg+5 μmol/L),with 12 rats in each group.Except for the CK group,all other groups were injected with doxorubicin through the tail vein to induce the NS rat model.Rats in CK group were injected with an equal volume of normal saline through the tail vein at the same time.After successful modeling,dosing treatment was performed once a day for 4 weeks.Detected 24-hour urine protein content,serum creatinine(Scr),albumin(ALB),urea nitro-gen(BUN)levels,renal tissue pathology,and levels of TNF-α,IL-6,and IL-1β in renal tissue;TUNEL staining was performed to detect cell apoptosis in rat kidney tissue;Western blot was performed to detect the expression of p-p38,p-JNK,p-ERK1/2 and p-NF-κB p65 proteins in rat kidney tissue.Results:Compared with CK group,Model group had severe renal tissue pathological damage,the 24 h urinary protein,Scr,BUN,TNF-α,IL-6,IL-1β,apoptosis rate,p-p38,p-JNK,p-ERK1/2,p-NF-κB p65 protein expressions were increased,while ALB level was decreased(P<0.05);compared with Model group,the renal tissue pathological damage of rats in Nef-L group,Nef-H group and PA group were severe,the 24 h urinary protein,Scr,BUN,TNF-α,IL-6,IL-1β,apoptosis rate,p-p38,p-JNK,p-ERK1/2,p-NF-κB P65 protein expressions were decreased,while ALB level was increased,the renal tissue pathological damage in the Anisomycin group was aggravated,the 24 h urinary protein,Scr,BUN,TNF-α,IL-6,IL-1β,apoptosis rate,p-p38,p-JNK,p-ERK1/2,p-NF-κB p65 protein expressions were increased,while ALB level was decreased(P<0.05);Anisomycin attenu-ated the effects of high doses of Nef on NS rats.Conclusion:Nef may alleviate the inflammatory injury in NS rats by inhibiting MAPK/NF-κB signaling pathway.

Objective:To discuss the clinical presentations,diagnosis,and treatment methods of the patients with Fournier's gangrene,and to enhance the clinicians'awareness of this condition.Methods:The clinical data including symptoms,signs,radiological findings,and surgical outcomes of one patient with Fournier's gangrene were collected.The relevant literatures were reviewed to summarize the clinical characteristics,diagnosis,and treatment methods for this condition.Results:The patient,a 42-year-old male,was admitted because of a history of infection around the perineum,scrotum,and perianal area for 13 d.His medical history included acute myeloid leukemia for 10 months,during which the patient underwent eight chemotherapy sessions in the local hospital.The abdominal CT scan results showed thickened,dense,and turbid soft tissue in the left inguinal area.The complete blood count reuslts showed the white blood cell count was 23.99×109 L-1.The cultures of wound secretions grew the Escherichia coli and Proteus mirabilis.The examination results showed there was necrosis of the scrotal skin and skin near the anus on the left buttock;the skin was blackened,hard,and demarcated from the surrounding normal skin with slight purulent exudation and no foul smell.The surrounding skin was significantly swollen and red;the rectal examination results showed no bleeding or fistulas.The patient underwent emergency debridement surgery on the admission day,followed by dressing changes,multiple applications of simplified negative pressure,perineal flap reconstruction,and skin grafting.The patient recovered well with normal function and had no complications.Conclusion:Fournier gangrene has acute onset and rapid progression,and the clinical manifestations are non-specific.The range of infection is not consistent with the progression of the disease.The diagnosis mainly depends on intraoperative exploration.Repeated radical surgery is the main treatment.The prognosis of this disease is good,and the recurrence rate is low,although long-term follow-up is still necessary after surgery.