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.

AIM To investigate the effects of stir-frying,processing with butter and carbonizing by stir-frying on oil components in Gleditsiae sinensis Fructus and Gleditsiae Fructus Abnormalis.METHODS The volatile oils and fatty oils were extracted by steam distillation method and Soxhlet extraction method,respectively,after which the extraction rates were determined.GC-MS was applied to analyzing the kinds and relative contents of oil components,after which cluster analysis was performed.RESULTS After the processing,the two medicinal materials demonstrated increased extraction rates of fatty oils and decreased extraction rates of volatile oils(except for processing with butter),the extraction rates of oil components in Gleditsiae sinensis Fructus were higher than those in Gleditsiae Fructus Abnormalis,and the reduced relative contents of toxic olefin benzene components were observable.CONCLUSION The kinds and relative contents of oil components in Gleditsiae sinensis Fructus and Gleditsiae Fructus Abnormalis exist obvious differences,the former displays better medicinal quality,whose processing mechanism in alleviating dryness and strength may contribute to the reduction of relative contents of toxic olefin benzene components.

Objective Viral encephalitis is an infectious disease severely affecting human health.It is caused by a wide variety of viral pathogens,including herpes viruses,flaviviruses,enteroviruses,and other viruses.The laboratory diagnosis of viral encephalitis is a worldwide challenge.Recently,high-throughput sequencing technology has provided new tools for diagnosing central nervous system infections.Thus,In this study,we established a multipathogen detection platform for viral encephalitis based on amplicon sequencing. Methods We designed nine pairs of specific polymerase chain reaction(PCR)primers for the 12 viruses by reviewing the relevant literature.The detection ability of the primers was verified by software simulation and the detection of known positive samples.Amplicon sequencing was used to validate the samples,and consistency was compared with Sanger sequencing. Results The results showed that the target sequences of various pathogens were obtained at a coverage depth level greater than 20×,and the sequence lengths were consistent with the sizes of the predicted amplicons.The sequences were verified using the National Center for Biotechnology Information BLAST,and all results were consistent with the results of Sanger sequencing. Conclusion Amplicon-based high-throughput sequencing technology is feasible as a supplementary method for the pathogenic detection of viral encephalitis.It is also a useful tool for the high-volume screening of clinical samples.

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.

To address the key challenges in multivariate statistical modeling,a higher-order derivative approach combined with vector space angle multiplicative error correction was proposed for establishing an acid value prediction ordinary least squares(OLS)regression model based on attenuated total reflectance-Fourier transform infrared(ATR-FTIR)spectroscopy.By using acid values measured by potentiometric titration as reference,ATR-FTIR spectroscopy was utilized for direct calibration and prediction of acid values on 96 kinds of lubricating oil samples from a wind turbine.Firstly,the simulated hyperbolic(SH)method was employed to obtain accurate fourth derivative spectrum,resolving overlapping bands and enhancing spectral selectivity.Then,from the calibration set(48 samples),informative spectral regions were identified based on correlation coefficients.Next,the sample with the highest acid value was selected as the reference and1/(1+tan(θ/2))was used as the metric relation of the spectrum to suppress the multiplicity error caused by factors such as the change of effective optical path in ATR-FTIR spectroscopy.After pretreatment of the spectrum by the method of fourth-order derivative combined with angular quantity,the number of variables decreased from 1737 to 8,and the matrix condition number decreased from 1.85×1015 to 56.34,which effectively eliminated the collinearity issue for OLS regression.Direct OLS modeling on spectral preprocessed data achieved a determination coefficient of 0.981 for 47 validation samples,with a relative error range of-8.38%-8.22%,outperforming the commonly used partial least squares(PLS)method(Determination coefficient of 0.865,relative error of-27.82%-22.38%).It was proved that effective data preprocessing significantly improved the prediction accuracy of the model.Furthermore,when the number of calibration set was compressed to 25 and the number of validation set was expanded to 70,the model retained 8 variables with a condition number of 42.60,the determination coefficient of validation set was 0.972,and the relative error ranged from-10.80%to 12.31%.Comparing with the PLS method(Determination coefficient of 0.724,relative error of-34.26%-53.84%),the improvement was more obvious,which showed that the method could still have high prediction accuracy even with fewer modeling samples as well as robustness against multiplicative error interference.

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.