Objective To understand the implementation of WS/T 508-2016 and the status of washing and disin-fection of medical textiles in China,and provide basis for the revision of the standard.Methods A questionnaire survey was conducted on the management of medical institutions and washing and disinfection workplace,building layout,personnel protection,equipment and supplies,washing and disinfection principles,and hygiene quality mo-nitoring of medical textiles in 323 medical institutions and 31 washing institutions in China.Meanwhile,microbio-logical index sampling was conducted on 234 pieces of medical textiles in 9 medical institutions and 8 washing insti-tutions in Hubei,Shanxi,Shanghai,and Sichuan Provinces before and after washing and disinfection.Results The awareness rates of WS/T 508-2016 among medical institutions and washing institutions were 96.90％and 96.77％,respectively,and the implementation rates were 94.12％and 96.77％,respectively.47.99％medical in-stitutions use purchasing services for washing and disinfecting medical textiles,and the higher the level of the medi-cal institution was,the higher the proportion of purchasing services was(x2=15.312,P＜0.001).85.16％medi-cal institutions have conducted risk assessments on service providers,and 52.99％were responsible for or participa-ted in pre-job training by the medical institution.Washing institutions were higher than medical institutions in terms of system soundness rate,pre-job training rate,proportion of quality management leaders and full-time(part-time)quality inspectors,setting rate of hand-washing facilities in zones,passages,isolation barriers,dressing(buffer)room,and toilets,configuration rate of hygiene isolation washing equipment,tunnel washing unit,washing equip-ment with heating functions,drying equipment and mechanical ventilation facilities,and specialty machine washing rate.Among 234 medical textiles specimens,11.97％were detected fungi,and the total fungal colonies in 5 clean textiles exceeded 100 CFU/100 cm2.Conclusion WS/T 508-2016 can further standardize the washing and disin-fection of medical textiles in China through strengthening institutional management,improving the supervision level of service providers,perfecting hardware facilities and layout,introducing new technologies,and increasing the mo-nitoring on fungi in clean textiles.

Objective:To analyze the drug-resistance pattern and molecular epidemiological characteristics of Enterobacteriales carrying the blaNDM gene in Lishui, aiming to guide clinical anti-infection treatment. Methods:Non-duplicate blaNDM-carrying Enterobacteriales, isolated from Lishui Central Hospital, were collected and identified by VITEK MS. The minimal inhibitory concentrations (MICs) were detected by the broth microdilution method. The ST types of the strains were determined by multilocus sequence typing (MLST). Plasmid types were identified by transformation or conjugation experiments and replication initiator amplification experiments. The transposon structures were detected by PCR amplification. Finally, the epidemic regularity of blaNDM gene in Lishui was analyzed from three levels: clonal group, plasmid, and mobile genetic elements. Results:A total of 109 blaNDM-positive strains were collected. Among them, 60 strains carried the blaNDM-1 gene and 49 strains carried the blaNDM-5 gene. The 109 strains showed 100% resistance to ceftazidime and cefotaxime. The resistance rates to peracillin-tazobactam and imipenem were higher than 80%. Strains carrying the blaNDM-5 gene were more resistant to meropenem than those carrying blaNDM-1 gene( P<0.05). A total of 68 STs were detected from 109 strains, and IncX3, IncFⅡγ, IncA/C and IncT/R plasmids were detected, and 90.83% of the blaNDM genes were located in the IncX3 plasmid. Twelve types of blaNDM gene surrounding structures existed, and they all carried the highly conserved blaNDM- bleMBL- trpF gene sequence. Conclusions:The blaNDM gene has diverse transmission modes in Lishui. The IncX3 plasmid is the main factor mediating its transfer, and all strains carry highly conserved blaNDM- bleMBL- trpF gene sequence.

Objective To understand the implementation of WS/T 508-2016 and the status of washing and disin-fection of medical textiles in China,and provide basis for the revision of the standard.Methods A questionnaire survey was conducted on the management of medical institutions and washing and disinfection workplace,building layout,personnel protection,equipment and supplies,washing and disinfection principles,and hygiene quality mo-nitoring of medical textiles in 323 medical institutions and 31 washing institutions in China.Meanwhile,microbio-logical index sampling was conducted on 234 pieces of medical textiles in 9 medical institutions and 8 washing insti-tutions in Hubei,Shanxi,Shanghai,and Sichuan Provinces before and after washing and disinfection.Results The awareness rates of WS/T 508-2016 among medical institutions and washing institutions were 96.90％and 96.77％,respectively,and the implementation rates were 94.12％and 96.77％,respectively.47.99％medical in-stitutions use purchasing services for washing and disinfecting medical textiles,and the higher the level of the medi-cal institution was,the higher the proportion of purchasing services was(x2=15.312,P＜0.001).85.16％medi-cal institutions have conducted risk assessments on service providers,and 52.99％were responsible for or participa-ted in pre-job training by the medical institution.Washing institutions were higher than medical institutions in terms of system soundness rate,pre-job training rate,proportion of quality management leaders and full-time(part-time)quality inspectors,setting rate of hand-washing facilities in zones,passages,isolation barriers,dressing(buffer)room,and toilets,configuration rate of hygiene isolation washing equipment,tunnel washing unit,washing equip-ment with heating functions,drying equipment and mechanical ventilation facilities,and specialty machine washing rate.Among 234 medical textiles specimens,11.97％were detected fungi,and the total fungal colonies in 5 clean textiles exceeded 100 CFU/100 cm2.Conclusion WS/T 508-2016 can further standardize the washing and disin-fection of medical textiles in China through strengthening institutional management,improving the supervision level of service providers,perfecting hardware facilities and layout,introducing new technologies,and increasing the mo-nitoring on fungi in clean textiles.

Objective:To analyze the drug-resistance pattern and molecular epidemiological characteristics of Enterobacteriales carrying the blaNDM gene in Lishui, aiming to guide clinical anti-infection treatment. Methods:Non-duplicate blaNDM-carrying Enterobacteriales, isolated from Lishui Central Hospital, were collected and identified by VITEK MS. The minimal inhibitory concentrations (MICs) were detected by the broth microdilution method. The ST types of the strains were determined by multilocus sequence typing (MLST). Plasmid types were identified by transformation or conjugation experiments and replication initiator amplification experiments. The transposon structures were detected by PCR amplification. Finally, the epidemic regularity of blaNDM gene in Lishui was analyzed from three levels: clonal group, plasmid, and mobile genetic elements. Results:A total of 109 blaNDM-positive strains were collected. Among them, 60 strains carried the blaNDM-1 gene and 49 strains carried the blaNDM-5 gene. The 109 strains showed 100% resistance to ceftazidime and cefotaxime. The resistance rates to peracillin-tazobactam and imipenem were higher than 80%. Strains carrying the blaNDM-5 gene were more resistant to meropenem than those carrying blaNDM-1 gene( P<0.05). A total of 68 STs were detected from 109 strains, and IncX3, IncFⅡγ, IncA/C and IncT/R plasmids were detected, and 90.83% of the blaNDM genes were located in the IncX3 plasmid. Twelve types of blaNDM gene surrounding structures existed, and they all carried the highly conserved blaNDM- bleMBL- trpF gene sequence. Conclusions:The blaNDM gene has diverse transmission modes in Lishui. The IncX3 plasmid is the main factor mediating its transfer, and all strains carry highly conserved blaNDM- bleMBL- trpF gene sequence.

Objective:To explore and summarize the morphological features of laryngeal tuberculosis under electronic laryngoscopy and image-enhanced endoscopy (i-scan).Methods:A retrospective analysis was conducted on the data of 31 patients diagnosed with laryngeal tuberculosis at the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2014 to June 2024, encompassing the morphological features of electronic laryngoscopy and i-scan endoscopy, histopathological features, and supplementary clinical examination results. Descriptive statistical methods were employed for the analysis.Results:Thirty-one patients were diagnosed with laryngeal tuberculosis, including 25 males and 6 females, aged from 21 to 84 years old, with an average age of 57 years old, and the disease course was from 1 to 12 months with an average of 3 months. The clinical symptoms included hoarseness in 27 cases, foreign body sensation in the pharynx in 2 cases, and sore throat in 2 cases. Twenty-six cases involved a single site, including 24 cases of the vocal cords, 1 case of the ventricular bands, and 1 case of the epiglottis; 5 cases involved multiple sites, including 2 cases of the vocal cords and the interarytenoid area, 2 cases of the aryepiglottic fold and the epiglottis, and 1 case of the aryepiglottic fold, the ventricular bands and the epiglottis. Eighteen patients showed a single morphology type under electronic laryngoscopy, including 4 cases of the edematous exudative type, 5 cases of the ulcerative type, and 9 cases of the granulation hyperplasia type. Edema, ulcer, and granulation hyperplasia can coexist and transit between each other. A total of 13 cases presented with two or more morphological types, with the edematous exudative type, the ulcerative type, and the granulation hyperplasia type occurred 7, 9, and 10 times respectively. Twenty-two patients had active or chronic pulmonary tuberculosis, while 9 patients had normal lung imaging. The laryngeal tissue biopsy pathology of 21 patients was chronic granuloma, and 10 patients were chronic granuloma with caseous necrosis; among them, 19 cases completed the Ziel-Neelsen staining (7 cases were positive for acid-fast bacilli), and 3 cases completed the polymerase chain reaction (PCR) (All were positive). Twenty cases completed the T-cell assay for tuberculosis infection (19 cases were positive), 15 cases completed the sputum smear (6 cases were positive), 18 cases were tested for antinuclear antibody (6 cases were positive), and 14 cases completed the erythrocyte sedimentation rate (4 cases were positive). Six patients underwent i-scan examination. In cases of ulcerative laryngeal tuberculosis without granulation hyperplasia in the surrounding tissues, i-scan revealed an abundance of abundant slightly thickened and tortuous oblique and dendritic blood vessels around the ulcer. If the pseudomembrane in the deep ulcer was thick, the blood vessel shadow was not visible. In shallow ulcers, there were areas of deep congestion and scattered dot-shaped blood vessel shadowst with uneven distribution; in cases of laryngeal tuberculosis with coexistence of the edematous exudative type and the granuloma type, i-scan visualized laryngeal cord edema with white exudates on the surface. Beneath the laryngeal cord exudates, there were scattered dot-shaped and irregularly distorted linear blood vessel shadows with uneven distribution, and tortuous, oblique, and dendritic blood vessels were observed around the lesion. In granuloma-type laryngeal tuberculosis, i-scan demonstrated that the area of granulation hyperplasia around the ulcer focus was significantly congested, characterized by scattered thick dot-shaped blood vessel shadows and irregularly distorted linear blood vessel shadows.Conclusions:Laryngeal tuberculosis presents as the edematous exudative type, the ulcerative type, and the granulation hyperplasia type under electronic laryngoscopy, and these types can coexist and interact. i-scan endoscopy can reveal detailed microvascular morphology and other subtle morphological characteristics. The identification and summary of these morphological characteristics are beneficial for the early detection and diagnosis of laryngeal tuberculosis.

Objective To compare the success rates of different culture methods for hemorrhagic amniotic fluid.Methods Thirty-one hemorrhagic amniotic fluid samples from pregnant women who were subjected to chromosomal examination at Anhui Maternal and Child Health Hospital were collected from January 2021 to December 2022.Two culture methods,the slide in situ culture box method(re-ferred to as the slide method)and the plastic bottle culture combined with slide in situ culture box method(referred to as the combined culture method),were used for cell cultivation.All the cells were harvested and stained with Giemsa staining,then the number of eligi-ble karyotype was counted and the success rates were compared between the two methods.Results Among the 31 cases of the slide method,21 were successfully cultured with a success rate of 67.7％.For the combined culture method,all the 31 cases were success-fully cultured with a success rate of 100％.The success rate of the combined culture method was significantly higher than that of the slide method(P＜0.05).Of 31 bloody amniotic fluid samples,three cases were fresh bleeding,and an average number of eligible kary-otype was 8 in the slide method and 32 in the combined culture method.Twenty-eight cases were old bleeding,and an average number of eligible karyotype was 13 in the slide method and 53 in the combined culture method.The number of eligible karyotype in the com-bined culture method was significantly higher than that of the slide method(P＜0.05).Conclusion The combined culture method is suitable for the cultivation of hemorrhagic amniotic fluid samples,and should be worthy of promotion in the clinics.

Objective:To establish a rapid method to detect the minimum inhibitory concentration (MIC) of imipenem in Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae (KPC-Kp) based on ompK36 gene′s GD mutation. Methods:This was a methodological evaluation study. A total of 258 isolates of Klebsiella pneumoniae were collected from Lishui Municipal Central Hospital from March 2011 to December 2019. Porin gene ompK36 and carbapenemase genes blaKPC, blaNDM, blaIMP and blaOXA-48 were amplified by PCR and confirmed by sequencing. The MIC was detected and confirmed by microbroth dilution susceptibility test, and the corresponding patterns of genotype and MIC were constructed. Based on the patterns, a method for rapid detection of imipenem MIC by real-time fluorescence PCR (RT-PCR) was designed and established. The 159 isolates of non-repetitive Klebsiella pneumoniae collected by Lishui Disease Prevention and Control Center (CDC) from 2017 to 2019 were used for further verification. The sensitivity and specificity were calculated by fourfold table. Kappa test was used to compare the consistency between RT-PCR and microbroth dilution susceptibility test. Results:Among 258 isolates, 109 isolates did not carry carbapenemase gene, 65 isolates carried ompK36 gene GD mutation, 127 isolates carried blaKPC, 15 isolates carried blaNDM, 9 isolates carried blaIMP, and blaOXA-48 was not detected. With mircobroth dilution susceptibility test as the standard, there were 3 corresponding patterns between the drug resistance gene and the imipenem MIC of Kp: when all the 4 carbapenemase genes were negative, MIC≤1 mg/L, the sensitivity was 100% (107/107) and the specificity was 98.4% (125/127); when blaKPC was positive and ompK36 gene GD mutation was negative, 4 mg/L≤MIC≤16 mg/L, the sensitivity was 88.2% (60/68) and the specificity was 98.8% (164/166); when blaKPC and ompK36 gene GD mutation were both positive, MIC≥32 mg/L, the sensitivity was 96.6% (57/59) and the specificity was 96.6% (169/175). RT-PCR detected blaKPC, blaNDM, blaIMP, blaOXA-48 genes accurately.The RT-PCR results of ompK36 gene GD mutation in the KPC-producing isolates were 100% consistent with the sequencing results. In the 159 isolates from Lishui CDC, the sensitivity and specificity of imipenem MIC detected by RT-PCR were higher than 95% in all 3 patterns with mircobroth dilution susceptibility test as the standard, and Kappa value was 0.971. Conclusion:The RT-PCR based on ompK36 gene GD mutation was helpful to quickly determine the MIC range of imipenem in KPC-Kp.

BACKGROUND:There is an internal relationship between hyperhomocysteinemia and vascular calcification.However,the pathogenesis of hyperhomocysteinemia promoting vascular calcification is still unclear. OBJECTIVE:To investigate the role of bone morphogenetic protein-2 in hyperhomocysteinemia-induced vascular calcification. METHODS:Human carotid wax samples were divided into a calcified group(n=29)and a non-calcified group(n=13)according to the presence or absence of calcified plaque.Sixteen ApoE-/-mice were randomly divided into a control group and a hyperhomocysteinemia group,with 8 mice in each group.Bone morphogenetic protein-2 vector was used to transfect rat thoracic artery smooth muscle A7r5 cells,and gradient concentration of homocysteine(50,100,200,and 400 μmol/L)was utilized to treat A7r5 cells.Calcification was detected by alizarin red staining and hematoxylin-eosin staining.The interaction of bone morphogenetic protein 2 with Runt-related transcription factor 2 was detected by immunofluorescence,and the expressions of bone morphogenetic protein 2,Runt-related transcription factor 2,and α-smooth muscle actin were detected by immunohistochemistry and western blot assay. RESULTS AND CONCLUSION:(1)Human carotid artery tissue staining revealed that compared with the non-calcification group,inflammatory cells increased and calcification positive rate increased in the calcification group(P<0.05).Compared with the non-calcification group,the expressions of bone morphogenetic protein-2 and Runt-related transcription factor 2 were up-regulated,and the expression of α-smooth muscle actin was decreased in the calcification group(all P<0.05).(2)The staining of mouse arterial specimens exhibited that,the positive rate of calcified area in the hyperhomocysteinemia group was significantly higher than that in the control group(P<0.05);serum homocysteine level in the hyperhomocysteinemia group was significantly higher than that in the control group(P<0.05).Compared with the control group,the expressions of bone morphogenetic protein-2 and Runt-related transcription factor 2 were up-regulated,and the expression of α-smooth muscle actin was decreased in the hyperhomocysteinemia group(all P<0.05).(3)A7r5 cell culture analysis demonstrated that with the increase of homocysteine concentration gradient,the degree of calcification,the content of bone morphogenetic protein-2 and Runt-related transcription factor 2 protein in A7r5 cells increased(P<0.05),and the content of α-smooth muscle actin protein decreased(P<0.05).(4)The A7r5 cell culture analysis of overexpressed bone morphogenetic protein 2 showed that the calcification degree of the overexpressed bone morphogenetic protein 2 group was increased compared with the corresponding control group,the β-sodium glycerophosphate group,and the homocysteine group.RUNt-related transcription factor 2 expression up-regulated(P<0.05)and α-smooth muscle actin expression down-regulated(P<0.05).(5)The expression of bone morphogenetic protein 2 increased in A7r5 cells cultured with homocysteine in calcified medium,and the expression of Runt-related transcription factor 2 increased with the increase of bone morphogenetic protein 2 expression.(6)The results confirm that bone morphogenetic protein-2 is a key target gene in the regulation of smooth muscle cell phenotypic transformation resulting in vascular calcification by hyperhomocysteinemia.Targeted regulation of bone morphogenetic protein-2 reduces hyperhomocysteinemia-induced vascular calcification.

Objective:To explore and summarize the morphological features of laryngeal tuberculosis under electronic laryngoscopy and image-enhanced endoscopy (i-scan).Methods:A retrospective analysis was conducted on the data of 31 patients diagnosed with laryngeal tuberculosis at the Second Affiliated Hospital of Zhejiang University School of Medicine from January 2014 to June 2024, encompassing the morphological features of electronic laryngoscopy and i-scan endoscopy, histopathological features, and supplementary clinical examination results. Descriptive statistical methods were employed for the analysis.Results:Thirty-one patients were diagnosed with laryngeal tuberculosis, including 25 males and 6 females, aged from 21 to 84 years old, with an average age of 57 years old, and the disease course was from 1 to 12 months with an average of 3 months. The clinical symptoms included hoarseness in 27 cases, foreign body sensation in the pharynx in 2 cases, and sore throat in 2 cases. Twenty-six cases involved a single site, including 24 cases of the vocal cords, 1 case of the ventricular bands, and 1 case of the epiglottis; 5 cases involved multiple sites, including 2 cases of the vocal cords and the interarytenoid area, 2 cases of the aryepiglottic fold and the epiglottis, and 1 case of the aryepiglottic fold, the ventricular bands and the epiglottis. Eighteen patients showed a single morphology type under electronic laryngoscopy, including 4 cases of the edematous exudative type, 5 cases of the ulcerative type, and 9 cases of the granulation hyperplasia type. Edema, ulcer, and granulation hyperplasia can coexist and transit between each other. A total of 13 cases presented with two or more morphological types, with the edematous exudative type, the ulcerative type, and the granulation hyperplasia type occurred 7, 9, and 10 times respectively. Twenty-two patients had active or chronic pulmonary tuberculosis, while 9 patients had normal lung imaging. The laryngeal tissue biopsy pathology of 21 patients was chronic granuloma, and 10 patients were chronic granuloma with caseous necrosis; among them, 19 cases completed the Ziel-Neelsen staining (7 cases were positive for acid-fast bacilli), and 3 cases completed the polymerase chain reaction (PCR) (All were positive). Twenty cases completed the T-cell assay for tuberculosis infection (19 cases were positive), 15 cases completed the sputum smear (6 cases were positive), 18 cases were tested for antinuclear antibody (6 cases were positive), and 14 cases completed the erythrocyte sedimentation rate (4 cases were positive). Six patients underwent i-scan examination. In cases of ulcerative laryngeal tuberculosis without granulation hyperplasia in the surrounding tissues, i-scan revealed an abundance of abundant slightly thickened and tortuous oblique and dendritic blood vessels around the ulcer. If the pseudomembrane in the deep ulcer was thick, the blood vessel shadow was not visible. In shallow ulcers, there were areas of deep congestion and scattered dot-shaped blood vessel shadowst with uneven distribution; in cases of laryngeal tuberculosis with coexistence of the edematous exudative type and the granuloma type, i-scan visualized laryngeal cord edema with white exudates on the surface. Beneath the laryngeal cord exudates, there were scattered dot-shaped and irregularly distorted linear blood vessel shadows with uneven distribution, and tortuous, oblique, and dendritic blood vessels were observed around the lesion. In granuloma-type laryngeal tuberculosis, i-scan demonstrated that the area of granulation hyperplasia around the ulcer focus was significantly congested, characterized by scattered thick dot-shaped blood vessel shadows and irregularly distorted linear blood vessel shadows.Conclusions:Laryngeal tuberculosis presents as the edematous exudative type, the ulcerative type, and the granulation hyperplasia type under electronic laryngoscopy, and these types can coexist and interact. i-scan endoscopy can reveal detailed microvascular morphology and other subtle morphological characteristics. The identification and summary of these morphological characteristics are beneficial for the early detection and diagnosis of laryngeal tuberculosis.

Objective:To establish a method for determintation of chlorogenic acid and linarin in Yejuhua granules by HPLC.Methods:We applied HPLC methods. The Kromasil 100-5 C18 column (250 mm×4.6 mm,5 μm) was used, the mobile phase was acetonitrile-0.4%H 3PO 4 solution (gradient elution), the flow rate was 1.0 ml/min, the dection wavelenghth was 334 nm and the column temperture was 32 ℃. Results:Chlorogenic acid and buddleoside had good linearity in the ranges of 0.30-1.50 μg ( r2=0.999 1) and 0.12-0.62 μg ( r2=0.999 8), respectively. The average recoveries were 99.70% and 96.67%, with RSD<2%, respectively. Conclusion:The method is simple, rapid, reliable, efficient, and can be used for determination of chlorogenic acid and buddleoside in Yejuhua Granules.