Objective To analyze the current status of anti-bacterial activity of carbapenem-resistant Klebsiella pneumoniae and Escherichia coli clinically isolated from hospitalized patients,detect their related resistance genes,and provide reference for the clinical treatment of carbapenem resistant Enterobacteria(CRE)infections and the rational use of antibiotics.Methods A total of 400 non-repetitive isolates of Klebsiella pneumoniae and Escherichia coli isolated from clinical specimens of Punan Branch of Renji Hospital,Shanghai Jiao Tong University School of Medicine from January 2022 to December were collected.The minimum inhibitory concentrations of these strains against commonly used antibiotics were determined by the broth microdilution method.The carbapenemase and related resistance genes of CRE were detected by drug resistance phenotype testing and PCR.Results Among the 400 strains,51 strains were identified as CRE,accounting for 12.75%.Among these,49 strains produced carbapenemases,with 41 strains(80.39%)being CR Klebsiella pneumoniae and 10 strains(19.61%)being CR Escherichia coli.Among the CRE strains,34 strains(66.67%)carried blaKPC,13 strains(25.49%)carried blaNDM,and 2 strains(3.92%)carried blaOXA-48.Conclusion Compared with other commonly used antibiotics,colistin and tigecycline exhibited good in vitro antibacterial activity against carbapenemase-producing Klebsiella pneumoniae and Escherichia coli.In addition,there was good concordance between drug resistance phenotype testing and genotyping.Clinical microbiology laboratories could continuously monitor the drug resistance phenotype and genotype of CRE and develop appropriate treatment plans based on actual conditions.

Objective:To investigate the efficacy and safety of daratumumab in the treatment of systemic light chain amyloidosis.Methods:The clinical data of 24 patients with systemic light chain amyloidosis who received daratumumab-based regimens in Sichuan Provincial People's Hospital from January 2020 to November 2022 were retrospectively analyzed. The treatment process of patients was summarized and the therapeutic efficacy was evaluated. Kaplan-Meier method was used to make survival analysis and the adverse reactions were analyzed.Results:All 24 patients included 2 cases (8.33%) of Mayo 2004 stageⅠ, 2 cases (8.33%) of Mayo 2004 stage Ⅱ and 20 cases (83.33%) of Mayo 2004 stage Ⅲ. All patients were treated with daratumumab-based regimen, and 17 patients had evaluable efficacy. In the chemotherapy regimens, 15 patients received DVd (daratumumab + bortezomib + dexamethasone) regimen, 7 patients received DVCd (daratumumab + bortezomib + cyclophosphamide + dexamethasone) regimen, 1 patient received DRd (daratumumab + lenalidomide + dexamethasone) regimen, and 1 patient received DTd (daratumumab +thalidomide + dexamethasone) regimen. After 1 course of daratumumab-based regimens in 17 cases with evaluable efficacy, the strict complete remission (sCR) rate was 41.18% (7/17), the overall response rate (ORR) was 88.24% (15/17). Among 17 patients who received daratumumab-based chemotherapy regimen as the first-line treatment, sCR rate of 11 cases with evaluable efficacy was 36.36% (4/11) after 1 course of treatment ORR was 90.90% (10/11). Among 5 relapsed/refractory patients, sCR rate of 4 cases with evaluable efficacy was 50.00% (2/4) after 1 course of treatment; ORR was 75.00% (3/4). Among 24 patients, renal involvement was found in 17 patients at the initial diagnosis. After 1 course of daratumumab-based chemotherapy regimen, ORR of 7 cases with evaluable efficacy was 85.71% (6/7), among which 42.86% (3/7) patients with renal involvement had an assessed renal response of very good partial remission (VGPR) or above. At the initial diagnosis, 19 cases had cardiac involvement; ORR of 14 cases with evaluable efficacy was 85.71% (12/14), among which 42.86% (6/14) patients had cardiac response to VGPR or above. After daratumumab-based chemotherapy regimen, the main adverse reactions were infusion-related adverse reactions, myelosuppression and infection, all of which were tolerated by the patients. The median follow-up time of 24 patients was 7.0 months (0.5- 16.5 months), the median progression-free survival time was 7.0 months (0.5-16.5 months) and the median overall survival time was 7.0 months (0.5-35.0 months).Conclusions:Daratumumab-based chemotherapy regimen has good efficacy and safety in the treatment of systemic light chain amyloidosis.

Objective To analyze the epidemiological characteristics of norovirus infectious diarrhea in the active monitoring of foodborne diseases in Jiangsu Province from 2015 to 2020, so as to provide reference for the prevention and control of foodborne diseases caused by norovirus. Methods Norovirus positive diarrhea cases were collected from sentinel hospitals in 13 districts and cities of Jiangsu Province, and their epidemiological and clinical characteristics were analyzed. Results Atotal of 3 620 norovirus positive cases were detected and isolated from 61 489 samples. The main serotype was GII (71.97%), the onset season was winter and spring, and the onset age was 1-3 years old and 14-34 years old. There was no significant difference in norovirus positive rate between different sexes, and the main symptom was diarrhea (incidence rate was 92.10%), Meat and meat products (20.20%) were the main types of suspected exposed foods. Conclusion Norovirus infection has obvious seasonal characteristics, and the population is generally susceptible. It is high in children and young people, and meat food was the main suspicious exposure food. We should continue to improve the ability of active monitoring, identification, early warning and control of foodborne diseases, so as to reduce the occurrence of foodborne diseases.

Objective To establish a method for the determination of 10 organophosphorus flame retardants in drinking water by on-line solid phase extraction coupled with high performance liquid chromatography-tandem mass spectrometry (On-line SPE-UPLC-MS/MS). Methods After adding the internal standard, the water sample was filtered by Millipore filtration, and then concentrated and detected by Online SPE-UPLC-MS/MS. Samples were concentrated by C₈ SPE column and separated by C₁₈ column with acetonitrile-water-formic acid as the mobile phases gradient elution，and were detected by multiple reaction monitoring (MRM) acquisition under anion mode. Results The 10 organophosphorus flame retardants all displayed good linear relationships within a certain range of concentrations, with the correlation coefficients being more than 0.990. The method detection limits were 0.60-5.50 ng/L, and the spiked recoveries of low, medium and high concentrations were 64%-106% , 83%-104% and 85%-99%, respectively. Conclusion The method is simple, sensitive, rapid, accurate and reliable, so it is applicable for the determination of 10 organophosphorus flame retardants in drinking water.

Objective To analyze the correlation between intestinal flora changes and neonatal necrotizing enterocolitis (NEC)through 16S rRNA metagenomic sequencing and bacterial culture. Methods From September 2018 to March 2019, 10 NEC cases and 6 controls were randomly selected in the neonatal ICU ward of Nanjing maternal and child health care hospital to analyze the 16S rRNA metagenomic diversity of the for intestinal flora. The fecal samples and corresponding environmental samples were corrected from 51 cases of NEC children and their case controls to isolate and culture Clostridium. Results The dispersion of samples within the case group was smaller than that of the control group, and the sample diversity was higher than that of the control group. In the isolation and culture of Clostridium, the overall detection rate of Clostridium in the case group was 43.14% (22/51), and the detection rate of Clostridium butyricum was the highest (19.61%, 10/51). There was a statistical difference between the two groups (χ²=5.85, P=0.015 58). All Clostridium strains did not carry the A, B and E type neurotoxin genes. Conclusion: Increased intestinal flora diversity, intestinal flora abundance and changes in the abundance of Clostridium may be closely related to the intestinal environment of children with NEC; Clostridium, especially Clostridium butyricum, may be related to the occurrence of NEC.

Objective To analyze the genotype-phenotype correlation in 5 families with congenital ectopia lentis (CEL) accompanied with cardiovascular abnormal manifestation.Methods Detailed clinical data of 15 family members in 5 families were collected from August 2017 to March 2018 in Zhongshan Ophthalmic Center,including examination of the condition of lens before and after mydriasis by slit-lamp,evaluation of the cardiovascular system using transthoracic echocardiography,and evaluation of the degree of involvement of the subjects' skeletal system using X-ray images.Genomic DNAs were extracted from whole blood sample of the 5 probands and 10 relatives,and screened for FBN1 mutation by targeted exome sequencing.The possible genotype-phenotype correlation was analyzed by reviewing previous literatures into these mutation sites.The study followed the principles of the Helsinki Declaration and written informed consent was obtained from each subject prior to any examination.Results All of the five probands were diagnosed as CEL accompanied with cardiovascular abnormal manifestation.FBN1 gene mutations were identified in all of the five probands,including four missense mutations (c.2741G＞T,c.2585G＞T,c.1633C＞T,c.4260C＞G) and one splicing mutation (c.2114-1G＞C).It was predicted that all of the 5 mutations would alter the protein structure.Conclusions FBN1 gene has a high degree of clinical heterogeneity,and the early detection of ocular phenotypes combined with genetic screening is of great significance in the diagnosis of cardiovascular abnormalities.

Objective To compare the effect of novomix30 and metformin in treatment of diabetes mellitus combined with osteoporosis.Methods 117 patients diagnosed as type 2 diabetes mellitus (T2DM) combined with osteoporosis were collected from Sep.2013 to Apr.2017 in our hospital.They were divided into two groups:metformin group (n=53),and novomix group (n=64) according to different therapies.General information of the two groups were comparable.The patients of metformin group reveived calcium,alendronate and metformin.The patients of novomix group received calcium,alendronate and novomix.The lumbar bone mineral density (BMD),greater trochanter BMD,femoral neck BMD,Ward triangle BMD,the serum levels of BGP,alkaline phosphatase (ALP),β-cross-linked C-telopeptide of type Ⅰ collagen(β-CTX),1,25-(OH)2D3,the levels of fasting plasma glucose (FPG),2h plasma glucose (2h PG),HBA1c and adverse reactions were recorded during the treatment.Results The lumbar BMD,greater trochanter BMD,femoral neck BMD,Ward triangle BMD of novomix group were higher than those of metformin group after treatment (P＜0.05).The serum β-CTX levels of novomix group were lower than those of metformin group after treatment (P＜0.05),while the serum BGP,ALP,1,25-(OH)2D3 levels of novomix group were higher than those of metformin group after treatment (P＜0.05).The levels of FPG,2hPG,HBA1c had no significant difference between the two groups after treatment (P＞0.05).The adverse reactions rate of metformin group and novomix group was 28.30％ and 25.00％ respectively,and there was no significant difference between the two groups (P＞0.05).Conclusion Novomix30 can significantly improve bone metabolism level for patients combined with T2DM and osteoporosis,maintain bone remodeling balance,reduce bone loss,and improve BMD,which is worthy to be promoted.

Objective:To detect the effect of microRNA-7 ( miR-7 ) knockdown on pathology in murine acute lung injury ( ALI) model,and preliminarily explore its significance.Methods:Murine ALI model was performed by intraperitoneal injection of Li-popolysaccharide (LPS) (10 mg/kg) into miR-7KD mice and wild-type (wild type,WT) mice respectively.Then,the pathologic injury of lung tissue were observed by HE staining.And total cell count of bronchoalveolarlavage(BAL) was calculated.The relative expression of related cytokines in lung tissue was analyzed by Real-time PCR assay.Furthermore,the changes on proportion of innate immune cells (γδT cell and F4/80 macrophages cell) and adaptive immune cell ( CD4+T cell and CD8+T cell) were analyzed by FACS.Meanwhile, the expression of CD62L and CD69,as well as the absolute number,in CD4+T cell were also analyzed.Results: Compared with WT mice,pathological damage in lung tissues was significantly alleviated in miR-7KD mice.Real-time PCR analysis showed that the relative expression of IL-6 was obviously reduced (P<0.01),conversely,relative expression of IL-4 and TGF-βwere obviously increased (P<0.05).Furthermore,the total cell number in BAL also reduced significantly (P<0.05).Importantly,FACS analysis showed that the proportion and the absolute number of F4/80+Mφcells obviously reduced (P<0.05);however,the proportion of γδT cells increased (P<0.05).Moreover,the proportion and the absolute number of CD4+T cells and CD8+T cells were significantly reduced (P<0.05). Finally, the proportion and the absolute number of CD62L+in CD4+T cells were upregulated vigorously,contrastly,the proportion and the absolute number of CD69+in CD4+T cells were notably up-regulated (P<0.05).Conclusion:miR-7 defeciency could significantly ameliorate the pathology of murine ALI,suggesting that it may play an important regulatory role in the development of ALI.

OBJECTIVE:To optimize the extraction and purification technology of total flavonoids from Engelhardia roxburghi-ana,and to establish the method for the content determination of 3 kinds of effective components. METHODS:Using the extrac-tion transfer rate of astilbin as index,single factor test was used to investigate extraction solvent,extraction method,volume frac-tion of percolation solvent ethanol,percolation material-liquid ration,soaking time before percolation and percolation rate of extrac-tion technology,and volume fraction of eluant ethanol in AB-8 resin purification technology. The contents of 3 effective compo-nents as astilbin,texifolin and engelitin in total flavonoids from E. roxburghiana were determined by HPLC. RESULTS:The opti-mal extraction technology was using 70% ethanol as extraction and percolation solvent,percolation extraction,soaking for 8 h be-fore percolation,percolation material-liquid ratio of 1∶16(g/ml),percolation rate of 30 ml/(min·kg). The purification technology was diluting the solution to 0.5 g (crude drug)/ml with water,ethyl acetate extraction,dissolved extract with 50% ethanol after evaporated to dryness,AB-8 resin for sampling,eluted with 50% ethanol,concentrating and drying. In verification test,extraction transfer rate of astilbin was more than 80%(RSD=0.42%,n=3). The contents of astilbin,taxifolin and engeletin in total flavo-noids from E. roxburghiana by purified were 57.94%,3.72% and 2.83%,respectively;the contents of 3 components accounted for 64.00% of total flavonoids. CONCLUSIONS:The extraction and purification technology is stable,rational and reliable;the content determination method of 3 effective components in total flavonoids of E. roxburghiana is accurate,simple and producible.

Objective:To detect the change of proportions of αβT lymphocytes in mesenteric lymph nodes of miR-7 knockdown (miR-7KD)mice,and preliminarily explore its importance.Methods: The volume,weight index and total cells number of mesenteric lymph node in miR-7KD mice were measured.The pathologic morphology change of mesenteric lymph nodes was observed by HE stai -ning.And the changes on proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice were analyzed by Flow cytometry.Results:Compared with those of WT (wild type) mice,the volume,weight index,and the total cells number of mesenteric lymph node were significantly increased ( P<0.05 ).Moreover ,the pathologic morphology was significantly changed.The proportion and numbers of T lymphocyte were significantly increased;however,the proportion of B lymphocyte were significantly decreased (P<0.05). Notably,the proportion and number of CD4+T cells and CD8+T cells were significantly increased (P<0.05).Meanwhile,CD62L+T cell proportion were vigorously reduced and CD 69+T cell and IFN-γ+T cell proportion were notably up-regulated,which belong to CD4+and CD8+T cell(P<0.05).Conclusion:There was significant influence in the proportion of αβT lymphocytes in mesenteric lymph nodes of miR-7KD mice,suggesting that miR-7 might play an important role in the composition and function of lymphocytes in mesenteric lymph nodes.