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.

The dental operative microscope has been widely employed in the field of dentistry, particularly in endodontics and operative dentistry, resulting in significant advancements in the effectiveness of root canal therapy, endodontic surgery, and dental restoration. However, the improper use of this microscope continues to be common in clinical settings, primarily due to operators' insufficient understanding and proficiency in both the features and established operating procedures of this equipment. In October 2019, Professor Jingping Liang, Vice Chairman of the Society of Cariology and Endodontology, Chinese Stomatological Association, organized a consensus meeting with Chinese experts in endodontics and operative dentistry. The objective of this meeting was to establish a standard operation procedure for the dental operative microscope. Subsequently, a consensus was reached and officially issued. Over the span of about four years, the content of this consensus has been further developed and improved through practical experience.
Humans ; Dentistry, Operative ; Consensus ; Endodontics ; Root Canal Therapy ; Dental Care

Diabetic retinopathy(DR)is a neurovascular unit lesion caused by diabetes mellitus,the incidence of which has increased significantly with the increase of the diabetic population.Therefore,it is important to use a reasonable animal model to explore the pathogenesis and therapeutic targets of DR.The db/db mouse is a relatively mature animal model of spontaneous type 2 diabetes and is therefore widely used in early prevention and treatment studies of type 2 DR.This paper describes the progress of db/db mouse model in exploring the pathogenesis of DR and the pharmacological research of Chinese and Western medicine,in order to provide a certain reference for the research of DR and drug development.

Objective To investigate the changing trend of platelet count (PLT) and related influencing factors in patients with hepatitis B virus-related chronic-on-acute liver failure (HBV-ACLF) after artificial liver support system (ALSS) therapy. Methods A total of 152 patients with HBV-ACLF who were hospitalized and treated in The Third Affiliated Hospital of Sun Yat-Sen University from January 2018 to November 2021 were included in the study, among whom 102 patients received plasma exchange (PE) and 50 patients received double plasma molecular absorption system combined with low-dose PE, and their clinical data and laboratory marker were measured. The independent samples t -test or the Mann-Whitney U test was used for the comparison of continuous data between two groups, and the chi-square test was used for the comparison of categorical data between two groups; a multivariate logistic regression analysis was used to investigate the risk factors for PLT > 50×10 9 /L after ALSS therapy; the receiver operating characteristic (ROC) curve was used to investigate the value of baseline PLT in predicting PLT > 50×10 9 /L after ALSS therapy. Results The patients were mostly middle-aged male adults; among the 152 patients, 70 (46.1%) had liver cirrhosis on admission, 114 (75.0%) received three sessions of ALSS therapy, and 88% had a baseline PLT count of > 50×10 9 /L. There was a significant reduction in PLT from baseline to after ALSS therapy (79.5±47.7 vs 112.5±64.1, t =4.965, P < 0.001), and at 1 week after treatment, PLT increased to the baseline level (97.2±50.7 vs 112.5±64.1, t =1.787, P =0.075). As for the change in PLT from baseline to 1 week after ALSS therapy, the liver cirrhosis group had a significantly greater reduction in PLT than the non-liver cirrhosis group ( U =1986.5, P =0.026), while there was no significant difference between different procedures of ALSS therapy and different sessions of treatment (3-5 sessions) (all P > 0.05). The multivariate logistic regression analysis showed that cirrhosis (odds ratio [ OR ]=3.097, 95% confidence interval [ CI ]: 1.255-7.645, P =0.014) and PLT > 50×10 9 /L at baseline ( OR =0.019, 95% CI : 0.002-0.154, P < 0.001) were independent risk factors for PLT > 50×10 9 /L after ALSS therapy. The ROC curve analysis of baseline PLT showed that PLT > 80.5×10 9 /L at baseline was the optimal cut-off value affecting PLT > 50×10 9 /L after treatment, with an area under the ROC curve of 0.818. Conclusion The influence of ALSS therapy on PLT is temporary, but cirrhotic patients have a weaker PLT generation ability than non-cirrhotic patients. PLT > 80.5×10 9 /L at baseline is the optimal cut-off value to reduce the risk of bleeding after ALSS therapy.

OBJECTIVE：To optim ize the ultrafiltration technology of enzymatic hydrolysate from Eucommia ulmoides peel. METHODS：The single factor test was adopted to investigate the effects of molecular weight of ultrafiltration membrane ，liquid temperature，operating pressure ，operating frequency ，membrane filtration time ，liquid concentration and pH on transfer rates of aucubin，geniposide and chlorogenic acid as well as solid removal rate in enzymatic hydrolysate from E. ulmoides peel. Setting the molecular cut off of fixed ultrafiltration membrane of 100 000，liquid concentration of 7 g/L，and pH value of 7，the ultrafiltration technology was optimized by Box-Behnken design response-surface methodology and validated with liquid temperature ，operating pressure，operating frequency and membrane passing time as factors ，using comprehensive scores calculated from transfer rates of aucubin，geniposide and chlorogenic acid as well as solid removal rate as indexes. RESULTS ：The optimal ultrafiltration technology of enzymatic hydrolysate from E. ulmoides peel was as follows as liquid temperature of 35 ℃，operating pressure of 0.5 MPa，operating frequency of 35 Hz and membrane passing time of 42 min. Results of validation tests showed that the comprehensive scores of the transfer rates of aucubin ，geniposide and chlorogenic acid as well as solid removal rate in enzymatic hydrolysate from E. ulmoides peel was 78.06％（RSD＝1.43％，n＝3），and its relative error with the predicted value （77.18％） was 1.14％. CONCLUSIONS ：The optimized ultrafiltration technology is stable and reliable ，and can be used for the ultrafiltration purification of enzymatic hydrolysate from E. ulmoides peel.

Objective To explore the value of neural thresholds in diagnosing and treating diabetic peripheral neuropathy.Methods A total of 42 type 2 diabetics and 21 healthy counterparts were recruited,and grouped according to the patients' illness duration and level of hemoglobin A1c (HbA1C).The neural thresholds of their median,ulnar and superficial peroneal nerves were measured.Results The neural thresholds increased with the disease's duration.The average neural threshold of those living with the disease for 10 years or more was significantly higher than that with a duration of less than 10 years.The nerve thresholds also increased with the level of hemoglobin A1c(HbA1C),and they were significantly higher in the group with an HbA1C level ≥9 than those in the group with HbA1C＜9.There was a positive linear correlation between the neural threshold of the ulnar and superficial peroneal nerves and the HbA1C level (P＜0.05).However,there was no significant correlation between the neural thresholds of the median nerves and the level of HbA1C (P＞0.05).There was also a positive linear correlation between the neural thresholds of the median,ulnar and superficial peroneal nerves and the duration of the disease (P＜0.01).Conclusion The neural threshold changes with the duration of diabetes and the level of HbA1C.Evaluating the peripheral nerve function of diabetics may have clinical utility.

Objective To investigate the effect of acidic tumor microenvironment on invasion and migration and its mechanism in glioma cells. Methods (1) The pH value of the medium was adjusted by acid-base titration. Human glioma cells U87 and U251 were cultured in the acid group and the normal group with pH values of 6.4 and 7.4, respectively; and 3 d after cultivation, the expressions of hypoxia-inducible factor-2α (HIF-2α) and CD44 were detected by Western blotting; Transwell assay was used to examine the invasion and migration of U87 and U251 cells; immunofluorescence was employed to examine the CD44 expression. (2) The U87 and U251 cells were divided into small interfering RNA (siRNA) -nonsense sequence group and siRNA-CD44-1 group, and the siRNA nonsense sequences and siRNA-CD44-1 interfering fragments were transfected by lipofectin-3000, respectively; three d after transfection, the migration and invasion abilities of cells from the two groups were detected by Transwell assay. (3) U87 and U251 cells were divided into acid group (cultured with a pH value of 6.4), blank control group, siRNA nonsense sequence group, siRNA-CD44-1 group, and siRNA-CD44-2 group; and cells from the later four groups were cultured with a pH value of 7.4; after culture for 4 d, the siRNA-nonsense sequence group, siRNA-CD44-1 group and siRNA-CD44-2 group were transfected with siRNA-nonsense sequences, siRNA-cd44-1 interfering fragments and siRNA-CD44-2 interfering fragments, respectively; three d after transfection, the expressions of CD44, N-Ca, Vimentin, and matrix metalloproteinase (MMP)-2 proteins in these 5 groups were detected by Western blotting. Results (1) As compared with the normal group, the expression levels of HIF-2α and CD44 in U87 and U251 cells of the acid group were significantly increased; both Transwell and invasion experiments showed that the number of transmembrane cells in the acid group was significantly larger than that in the normal group (P<0.05); immunofluorescence staining showed that the CD44 expression in acid group was significantly higher than that in normal group (P<0.05). (2) Both Transwell and invasion experiments showed that the number of transmembrane cells in the siRNA-CD44-1 group was significantly smaller than that in the siRNA nonsense sequence group (P<0.05). (3) Western blotting showed that the expression levels of CD44, N-Ca, Vimentin and MMP-2 in U87 and U251 cells of the blank control group, siRNA nonsense sequence group, siRNA-CD44-1 group, and siRNA-CD44-2 group were obviously decreased as compared with those in the acid group; the expression levels of CD44, N-Ca, Vimentin and MMP-2 in U87 and U251 cells of the siRNA-CD44-1 group and siRNA-CD44-2 group were obviously lower than those in the siRNA nonsense sequence group. Conclusion Acidic tumor microenvironment enhances the capabilities of invasion and migration of glioma cells through increasing CD44 expression.

Objective To study the influence of micro (miR)-325 in progression of glioma and its molecular mechanism by regulating transferrin receptor (TFRC) gene expression in glioma cells. Methods (1) Thirty-five glioma tissues and paired adjacent normal tissues were collected during surgical excision performed in our hospital from January 2015 to January 2018. The miR-325 and TFRC mRNA expression levels in the glioma tissues and paired adjacent normal tissues were detected by inverse transcription-quantitative PCR (RT-qPCR); the expression of miR-325 in glioma tissues of patients with different clinical characteristics and the survival curves of patients with low or high miR-325 expressions were compared. (2) RT-qPCR was used to examine the miR-325 expression in HA, U251, and U87 cell lines in vitro; the regulatory relations between miR-325 and its potential target gene TFRC in U251, and U87 cell lines were measured by luciferase report assay; miR-325 mimic and its negative control were transfected into U251 and U87 cell lines for 48 h, and then, the mRNA and protein expressions of TFRC were detected by RT-qPCR and Western blotting, respectively; control small interfering RNA (siRNA)+nonsense inhibitor, TFRC siRNA+nonsense inhibitor, and siTFRC+miR-325 inhibitor were transfected into U251 and U87 cell lines for 48 h, respectively, Western blotting was employed to detect the TFRC protein expression, cell proliferation was detected by CCK-8 assay, and cell invasion was detected by Transwell assay; pcDNA3.1 empty vector+nonsense sequence, TFRC pcDNA3. 1+nonsense sequence, TFRC pcDNA3.1+miR-325 mimic were transfected into U251 and U87 cell lines for 48 h, respectively, TFRC protein expression was detected by Western blotting, cell proliferation was detected by CCK-8 assay, and cell invasion was detected by Transwell assay. Results (1) As compared with those in the adjacent tissues, the miR-325 expression was significantly decreased and the TFRC mRNA expression was statistically increased in glioma tissues (P<0.05); the TFRC mRNA expression and miR-325 expression were negatively correlated in glioma tissues (P<0.05); as compared with patients with Karnofsky functional status scores≥80, patients with scores<80 had significantly decreased miR-325 expression; as compared with glioma tissues of WHO grading I-II, glioma tissues of grading III-IV had significantly decreased miR-325 expression (P<0.05); the survival rate of patients with low miR-325 expression was statistically lower than that of patients with high miR-325 expression (P< 0.05). (2) As compared with that in HA cells, the miR-325 expression was statistically down-regulated in U87 and U251 cells (P<0.05); in TFRC wild-type (TFRC WT) transfected cells, the miR-325 mimic group had significantly lower luciferase activity than the nonsense sequence group, while the miR-325 inhibitor group had significantly higher luciferase activity than the nonsense inhibitor group (P<0.05); as compared with those in the nonsense sequence group, the TFRC mRNA and protein expressions were statistically decreased in U87 and U251 cells of miR-325 mimic group; as compared with those in the control siRNA+nonsense inhibitor group, the TFRC protein expression and absorbance value were significantly decreased, and number of invasive cells was significantly smaller in the siTFRC+nonsense inhibitor group; and as compared with those in the siTFRC+nonsense inhibitor group, the TFRC protein expression and absorbance value were significantly increased, and number of invasive cells was significantly larger in the siTFRC+miR-325 inhibitor group (P<0.05); as compared with the pcDNA3.1 empty vector+nonsense sequence group, the TFRC protein expression and absorbance value were significantly increased, and number of invasive cells was significantly larger in the TFRC pcDNA3.1 +nonsense sequence group, and as compared with the TFRC pcDNA3.1+nonsense sequence group, the TFRC protein expression and absorbance value were significantly decreased, and number of invasive cells was significantly smaller in the TFRC pcDNA3.1+miR-325 mimic group (P<0.05). Conclusion The miR-325 expression is decreased in glioma cells and has a tumor suppressor effect; patients with low miR-325 expression have poor prognosis; miR-325 inhibits cancer cell progression by inhibiting the expression of the target gene TFRC.

Objective To compare the effects of propofol and sevoflurane on oxidative stress re-sponses in patients undergoing one-lung ventilation. Methods Eighty American Society of Anesthesiolo-gists physical statusⅠorⅡpatients with lung cancer, aged 42-53 yr, weighing 52-83 kg, scheduled for e-lective pulmonary lobectomy performed via a thoracoscope, were divided into 2 groups ( n=40 each) using a random number table method: group propofol and group sevoflurane. Propofol was intravenously infused at 4-10 mg·kg-1 ·h-1 in group propofol. In group sevoflurane, 1％-3％ sevoflurane was inhaled. Forced expiratory volume (FEV), forced vital capacity (FVC), FEV in first second (FEV1), FEV∕FVC, and maximal expiratory flow ( MEF) were measured at 24 h after operation. Blood samples were obtained from the median cubital vein for determination of the levels of plasma malondialdehyde ( MDA) , catalase ( CAT) and superoxide dismutase ( SOD) and expression of NOX2 and NOX4 subunits-containing NADPH oxidase, SP-D and CC16 ( by Western blot) . Results Compared with group sevoflurane, FEV, FVC, FEV1, FEV∕FVC and MEF were significantly increased, the activity of plasma SOD and CAT was increased, MDA con-centration was decreased, the expression of NOX2 and NOX4 subunits-containing NADPH oxidase and SP-D was down-regulated, and the expression of CC16 was up-regulated in group propofol (P<0. 05). Conclu-sion Propofol provides better efficacy in protecting lung function of patients undergoing one-lung ventilation when compared with sevoflurane, which is related to inhibiting oxidative stress responses.

Objective To investigate the regulating mechanism of Notch1 signaling pathway on the invasion and migration ofglioma initiating cells (GICs).Methods (1) Box-plotting was conducted to analyze the mRNA expression of Notch1 in normal brain tissue and glioblastoma tissue using Bredel Brain,Sun Brain and TCGA databases;Kaplan-Meier survival analysis was conducted to analyze the association between the prognosis of glioma patients with the expression of Hes1 in TCGA database;Heatmap was conducted to analyze the expression of Notch1 and CXCR4 in GICs and common cell line in GEO database.(2) Magnetic activated cell sorting was adopted to establish cell lines of U87 GICs and U251 GICs;immunofluorescence staining was used to detect expression of CXCR4 and Notch1.After the cell lines of U87 GICs and U251 GICs were divided into DMSO,shNC,MK0752 and shNotchl groups,the shNotch1 and shNC groups were transfected respectively with recombinant lentivirus of Notch1-shRNA and its control sequence while the MK0752 and DMSO groups were added respectively with MK-0752 of 80 nmol/mL and the same amount of DMSO.The protein expression of Notch1,CXCR4 and p-mTOR was detected by Westem blotting in the 4 groups.The capabilities of invasion and migration of the GICs were detected by Transwell assay in the shNotch1 and shNC groups.Results (1) The box-plotting showed the mRNA expression of Notch 1 in the glioblastoma tissue was significantly higher than in the normal brain tissue (P＜0.05).The Kaplan-Meier survival analysis showed that the life span ofglioma patients with high expression of Hes1 was significantly shorter than that of those with low expression of Hes1 (P＜0.05).Heatmaps showed that the expression levels of Notch1 and CXCR4 in GICs were higher than in the common cell line.(2) The immunofluorescence staining showed that Notch1 and CXCR4 were highly expressed and colocalized in cell lines of U87 GICs and U251 GICs.The Western blotting showed that the protein expression of Notch1,CXCR4 and p-mTOR in the cell lines of U87GICs and U251 GICs in the MK0752 and shNotch1 groups was lower than that in the DMSO and shNC groups.Transwell assay showed that the penetrating-membrane cells per visual field in the shNotch1group were significantly fewer than those in the shNC group (P＜0.05).Conclusion Notch1 signaling pathway can promote invasion and migration of GICs through regulating CXCR4 expression.