Objective To detect ESBLs in E.coli and K.pneumoniae with appropriate method.Methods The suspiciously produced ESBLs were detected by standard screen test among 110 E.coli and 84 K.pneumoniae. ESBLs possessing strains were confirmed by E test, single disc diffusion test with clavulanic acid or sulbatan, and two kinds of double disc synergy test. Results There was a significant difference between the detected rate of ESBLs in E.coli (16/26) and K. pneumoniae (4/20) by E test( P 0.05). The single disc diffusion test with clavulanic acid or sulbatan was applicable to confirm ESBLs in K. pneumoniae (12/20). Conclusions Single disc diffusion test is a sensitive, convenient and inexpensive method to confirm ESBLs in E.coli and K.pneumoniae in clinical laboratory.

Objects To observe the TEM gene characteristic of Acinetinbacter lwoffi strains which were resistant to most kind of antibiotics and to compare their sequences with that of Escherichia coli Methods Two strains of A lwoffi, JN18 and JN70,were isolated from sputum of patients′ who suffered from respiratory infection TEM, SHV, OXA, IMP and CTX M genes were tested by PCR TEM sequences of JN18 and JN70 were detected by ABI automated sequencer and were analysed by DNAStar software to compare the differences with E coli TEM genes that had been published in GenBank Results Detected sequences of A lwoffi JN18 and JN70 strains were 1012 bp and 887 bp, respectively They coded regions of 832 bp and 772 bp for TEM of the two strains that were 98 2% identical In other hand, there were 14 pair bases differently in TEM regions The TEM sequence of JN18 was 98 72% identical to that of E coli TEM on the average, which was over 99% to TEM1D, TEM 70, TEM 76, TEM 77 and TEM 95 of E coli The divergence of TEM genes was 1 26 between A lwoffi JN18 strains and E coli JN70 strain had higher identity (98 93%), which reached 99 5% to TEM1D, TEM1F and TEM84 of E coli As JN18 strain, TEM gene of JN70 had very small divergence (1 06 ) with E coli Conclusion JN18 and JN70 strains of A lwoffi isolated from patients′respiratory tract in Jinan shared most of sequences with E coli TEM76 and 84 However, there were many mutant sites in them

Objective To optimize the extraction process of anti-hepatic fibrosis water-soluble component from Carthamus tinctorius L. via the orthogonal method. Methods The influences of type and content of solvent,extraction time,and extraction frequency on processing were investigated by assessing the content of hydroxysafflor yellow A,yield of extraction and anti-hepatic fibrosis potency via the high performance liquid chromatography and enzyme-linked immunosorbent assay by L9(3)4 orthogonal test. Results The optimum extracting condition for the anti-hepatic fibrosis water-soluble component from carthamus tinctorius L. was as follows: extracting with 10 times the volume of 10% ethanol for twice, 0. 5 h in each. Conclusion The process is scientific and feasible, which serves as a guideline for the production of the anti-hepatic fibrosis water-soluble components.

OBJECTIVE: To investigate the expression and clinicopathological significance of the cell cycle regulators cyclin E, cyclin D1, p21, p16 in laryngeal carcinogenesis tissus. METHOD: The expression of cell cycle regulators were detected by flow cytometry method in 23 cases of polyps of vocal cord, 69 cases of laryngeal precancerous change and 33 cases of laryngeal squamous cell carcinoma (LSCC), which tissue was paraffin embedded, sliced, dewaxed, and prepared into the cell suspension, then fluorescently labeled by cyclin E, cyclin D1, p21 and p16. RESULT: In polyps of vocal cord, laryngeal precancerous change and LSCC, The positive expression rate of cyclin E and cyclin D1 were respectively 13.04%, 20.29D, 42.420 and 26.09%, 43.48% and 93.94%. The positive expression rate of p16 and p21 were respectively 61.90%, 40.98%, 14.28% and 47.62%, 23.81%, 26.23%. Those showed the positive expression rate of cyclin D1, cyclin E gradually decreased from vocal cord polyps, laryngeal precancerous change to LSCC, (P < 0.05, P < 0.01), while the positive expression rate of p21 and p16 gradually decreased (P < 0.01). CONCLUSION The abnormal expression of cell cycle regulatory factors is the molecular events of laryngeal carcinoma. High expression of positive regulatory factors cyclin D1 and cyclin E, and low expression of negative regulatory factors p16 and p21, which showed the imbalance of multiple positive and negative regulatory factors related with cell cycle play an important role in the occurrence of laryngeal cancer.
Adult ; Cyclin D1 ; metabolism ; Cyclin E ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p21 ; metabolism ; Female ; Flow Cytometry ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Male ; Middle Aged ; Oncogene Proteins ; metabolism

17 patients with maxillary antral pseudocyst underwent side wall fenestration with simultaneous implantation of 38 implants and the upper denture repair, follow-up observation was conducted after repair for 6 to 24 months (an average of 11. 4 months) . Good osseointegration was observed in all cases. No implant shedding or bone resorption occurred during the follow-up period, and the antrol pseudo cyst disappeared or decreased. Dental implantation simulataneously with maxillary sinus augmentation is effective in the management of maxillary implantation for the cases with antrol pseudocyst.

Background:Antibiotics can reduce the efficacy of immunotherapy for melanoma,non-small cell lung cancer and renal cell carcinoma by altering the intestinal microbiome,but their effect in the treatment of gastrointestinal cancer is unclear.Aims:To investigate the prognostic effect of antibiotics on PD-1/PD-L1 inhibitors in the treatment of colorectal cancer(CRC)patients.Methods:The clinicopathological data of 95 patients diagnosed with CRC and treated with immune checkpoint inhibitors(ICIs)from December 2018 to May 2022 at Renmin Hospital of Wuhan University were retrospectively analyzed,and the patients were divided into antibiotic group and control group according to whether antibiotics were used.Risk factors of progression free survival(PFS)and overall survival(OS)were analyzed by univariate and multivariate analysis.Results:Among the 95 patients with CRC,64 were male and 31 were female.The mean age was(60.46±11.82)years.PFS(10.9 months vs.24.8 months,P=0.032)and OS(14.5 months vs.35.5 months,P=0.008)were significantly decreased in antibiotic group than in control group,and one-year survival rate was 56.5%and 78.7%,respectively.Multivariate analysis showed that tumor location and antibiotic use were independent risk factors of PFS(P<0.05),while tumor location,antibiotic use and no radical surgery were independent risk factors of OS(P<0.05).Univariate and multivariate analyses showed that antibiotic administration route,dose were not risk factors of PFS(P>0.05);univariate analysis showed that antibiotic administration route was correlated with OS(P=0.045),however,multivariate analysis showed that antibiotic administration route,infection location were not risk factors of OS(P>0.05).Conclusions:Tumors located in colon and antibiotics applied within 30 days before and after the first dose of ICIs can shorten PFS and OS in CRC patients,while radical surgery can prolong the OS of CRC patients.

Objective:To analyze clinical phenotypes and pathogenic mutations of a patient with classic tuberous sclerosis complex.Methods:Clinical data was collected from a patient with classic tuberous sclerosis complex. Next-generation sequencing was performed to screen pathogenic gene variants, and Sanger sequencing to verify the mutations. Minigene plasmids were constructed and transfected into the human renal epithelial cell line 293T, and RNA was extracted for transcriptional analysis.Results:The patient clinically presented with recurrent epileptic seizures, facial angiofibroma, periungual fibroma, pulmonary lymphangioleiomyomatosis, renal angiomyolipoma and multiple osteosclerosis. Next-generation sequencing revealed a suspected pathogenic variant in the TSC2 gene in the patient. Sanger sequencing identified a heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene in the patient, but not in his parents or 100 unrelated healthy controls. Moreover, this mutation had not been previously reported. The minigene experiment showed changed mRNA sequence of the TSC2 gene in this patient with loss of the authentic splice site in exon 4 and insertion of a 74-bp intron, which shifted the splice site 90 bp downstream (r.336delins336+16_336+90) .Conclusion:The novel heterozygous mutation c.336_336+15delGGTAAGGCCCAGGGCG in exon 4 of the TSC2 gene can lead to aberrant splicing, and may contribute to tuberous sclerosis complex in this patient.

OBJECTIVE: To explore the genetic basis for a patient diagnosed with tuberous sclerosis complex (TSC). METHODS: Peripheral blood samples of the patient and his parents were collected for the extraction of genomic DNA. Next generation sequencing (NGS) was carried out to detect potential variant, and the result was verified by Sanger sequencing. RESULTS: The patient was found to harbor a heterozygous c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene. The same variant was not found in his unaffected parents and 100 unrelated healthy controls. Based on the American College of Medical Genetics and Genomics guidelines, the variant was predicted to be pathogenic (PVS1+PS2+PM2). CONCLUSION The novel c.1053delG (p.Glu352SerfsX10) frameshifting variant of the TSC2 gene probably underlay the TSC in this patient.
Genomics ; Heterozygote ; Humans ; Mutation ; Tuberous Sclerosis/genetics* ; Tuberous Sclerosis Complex 2 Protein/genetics*

Objective:To investigate the effect of liraglutide(LRG) on high glucose-induced oxidative stress injury in(H9c2) cardiomyocytes and its underlying mechanisms.Methods:A high glucose treatment was applied to H9c2 cells for 24 hours to establish an in vitro model of myocardial cell injury. Different concentrations of liraglutide(10, 100, 1000 nmol/L) were administered for intervention. Cell viability was evaluated using the CCK-8 assay, and changes in cell morphology were observed under an inverted microscope. After 24 hours of liraglutide(100 nmol/L) intervention following high glucose treatment, the levels of lactate dehydrogenase(LDH), superoxide dismutase(SOD), and malondialdehyde(MDA) in the cell supernatant were measured. RT-PCR and Western blotting were used to detect the mRNA and protein levels of silent information regulator factor 1(SIRT1) and forkhead box protein O1(FOXO1). Western blotting was also used to assess the acetylation level of FOXO1 protein. Small interfering RNA(siRNA) technology was employed to silence SIRT1 in H9c2 cells to confirm its role in the study. Results:Compared to the control group, the high glucose group showed decreased cell viability, cell structure damage, increased levels of LDH and MDA in the cell supernatant, decreased SOD levels, aggravated oxidative stress, decreased SIRT1 expression, and increased acetylation level of FOXO1(all P<0.05). Compared to the high glucose group, liraglutide intervention resulted in increased cell viability, improved cardiac cell morphology, reduced oxidative stress levels, increased SIRT1 expression, and decreased acetylation level of FOXO1(all P<0.05). When SIRT1 was downregulated, the protective effects of liraglutide were weakened(all P<0.05). Conclusions:Liraglutide has a protective effect against high glucose-induced oxidative stress injury in H9c2 cells, which may be associated with the upregulation of SIRT1 expression.

Objective To investigate antimicrobial resistance among nosocomial gram-negative bacilli in 2006.Methods About 987 consecutive and non-repetitive gram-negative bacilli were isolated from 10 teaching hospitals from Sep.to Dec.in 2006 in China.All of these isolates were sent to the central laboratory for reidentification and susceptibility testing.The minimal inhibitory concentration(MICs)of meropenem and other antibacterial agents were determined by agar dilution method.Results The activity of antibacterial agents against Enterobacteriaceae was as fol lows in descending order of susceptible rate: meropenem(susceptible rate 99.8%),imipenem(99.5%),piperacillin/tazobactam(91.3%),amikacin (89.3%),cefepime(83.8%),cefoperazone/sulbactam(79.7%),ceftazidime(74.7%),cefotaxime (57.7%),ceftriaxone(56.6%),ciprofloxacin(53.6%).The prevalence of extended-spectrum β-Iactamases(ESBL)was 59.0% in Escherichia coli,33.0%in Klebsiella pneumoniae and 8.0%in Proteus mirabilis.The most active agents against E.coli and K.pneumoniae were meropenem,imipenem(99.2%. 100%),piperacillin/tazobactam(90.8%-97.0%),and amikacin(83.8%-92.4%).Cefepime Was more active against K.pneumoniae than E.coli(85.4% vs.65.2%).Against E.cloacae,E.aerogenes and Citrobacter freundii,the most active agents were as follows in desecnding order:meropenem,imipenem (99.2%-100%),amikacin(85.2%-92.6%),cefepime(81.5%-85.9%),piperacillin/tazobactam (73.4%-87.2%),cefoperazone/sutbactam(65.6%-77.7%),and ciprofloxacin(53.1%-72.3%).The most active agents against Pseudomonas aeruginosa were amikacin(83.5%),followed by meropenem (79.1%),piperacillin/tazobactam(74.1%),and imipenem(70.9%).The most susceptible agents against Acinetobacter baumannii were imipenem(79.1%),meropenem(73.4%) and cefoperazone/ sulbaetam(54.7%).Mutiresistant A.baumannii increased up to 53.0%.The most active agents against Burkholderia cepacia were meropenem(73.3%),eeflazidime(73.3%),and piperacillin/tazobactam (62.2%).Conclusions Carbapenems remained very high activity against Enterobacteriaceae.Increasing resistance to 10 antimicrobials agents tested from A.baumanni and P.aeruginosa brought great concern.