Objective:To quantitatively analyze the clinical and drug resistance feature of diarrhea of adults patients in 2016 and 2019 induced by the Escherichia coli (diarrheagenic Escherichia coli, DEC), and to reveal the difference of DEC′s epidemiological features of before and after measuring to strengthen food hygiene and safety in Beijing. Methods:A total number of 3 408 patients with food-borne adult diarrhea were received diagnosis and treatment in the intestinal clinic department of Beijing Tongren Hospital in 2016 and 2019.There were 1 926 patients in 2016 and 1 482 in 2019, respectively. The clinical information of patient were entered into the intestinal early warning system and were carefully preserved. The clinical specimens (the stool samples) were isolated and the DECs were identified by culturing. The colony of DECs was identified by matrix assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry. Five pathogenic types of Escherichia coli were classified by multiplex PCR methods. The drug-susceptibility test was performed according to the standards of the American Society for Clinical and Laboratory Standardization in 2019. The categorical data were analyzed by χ 2 test or Fisher′s exact test to verify the statistical difference. Results:A total number of 581 DECs strains were detected in 3 408 specimens. Among the subtypes of E Coli, the Enterotoxigenic Escherichia coli (ETEC) accounted for 53.36% (310/581), and Enterohemorrhagic Escherichia coli (EHEC) was detected. In 2016, the total detection rate of DEC was 14.54% (280/1 926), enteroaggregative Escherichia coli (EAEC) accounted for 18.21% (51/280), and ETEC accounted for 71.79% (173/280). In 2019, the total detection rate of DEC was 20.31% (301/1 482), EAEC accounted for 41.23% (116/301), and ETEC accounted for 48.93% (137/301). Compared with 2016, the detection rate of EAEC in 2019 increased significantly (χ2=29.26, P<0.001), followed by EPEC (χ2=9.37, P=0.002), and ETEC decreased (χ2=15.43, P<0.001). Compared with other pathogenic types, EAEC can easily cause nausea(χ2=32.72, P<0.001).The red blood cells(χ2=16.44, P=0.001) or the white blood cells (χ2=26.82, P<0.001) could be easily observed in stool specimens of patients infected with enteroinvasive Escherichia coli (EIEC). The resistance rates of EIEC to ampicillin, ampicillin/sulbactam and gentamicin were 80.95% (17/21), 66.67% (14/21) and 57.14% (12/21), respectively. Three strains of EAEC resistant to carbapenem antimicrobials were discovered in 2019 and of which two strains were resistant to ertapenem and imipenem, and the other one strain was only resistant to ertapenem. The whole genomic sequencing showed that there are multiple resistance mechanisms: including the mainly drug-resistant nodular cell differentiation family efflux pump, penicillin binding site mutation, and New Delhi metal-β-lactamase 5 production. Conclusions:The detection rate of DECs in adult patients with food-borne diarrhea is high, and the foremost subtype of DECs is ETEC. Compared with 2016, the detection rates of ETEC in clinical specimens decreased in 2019, and the detection rate of EAEC increased significantly, respectively. In 2019, a carbapenem-resistant antibacterial drug-resistant Escherichia coli strain was isolated. It is of great significance to focus on the biological characteristics and epidemiological changes of DEC.

Objective To probe the immunological traits of mesenchymal stem cells derived from umbilical cord Wharton's jelly (WJMSCs). Methods The diced Wharton's jelly which was from healthy fullterm birth human umbilical cord was cultured. The mesenchymal stem cells were identified with mesenchymal stem cells markers expression by flow cytometry and multiple differentiation ability. The expression of MHC- Ⅰ / Ⅱ, costimulatory molecules (CD40, CD80 and CD86) was detected with flow cytomctry, immunocytochemistry, and RT-PCR. The expression of immune inhibitors like HLA-G, IDO, and PGE2 was detected by immunocytochemistry and RT-PCR. The expression of immune-related molecules as IL-10, TGF-β, FGF and VEGF was detected with antibody microarray and western blot. Further more, to clarify the in vivo immune reaction of hWJMSCs, we fabricated the hWJMSC-scaffold constructs and implanted them into the rabbit backs. The lymphocyte infiltration and implanted cell survival observed with immunofluorescence. Results After culturinge of diced Wharton's jelly tissue, we obtained spindle-shaped cells. With differentiation medium, the cells can differentiate into osteoblasts, chongdrocytes, adipose cells and schwann cells. Expression of MHC, costimulatory molecules, and a series of immune suppressive-related molecules was found. Immune inhibitors as HLA-G, 1DO, PGE2, and immune suppressive related molecules as HGF, VEGF, TGFand IL-10 were positively expressed. But the cells did not express MHC-Ⅱ. No immune rejection was observed in vivo after implantation of hWJMSC-scaffold constructs. Conclusion It can be concluded that hWJMSCs have very low immunogenicity, which means the cells have potential to induce immune tolerance.The hWJMSCs do not provoke immune rejection in vivo.

AIMTo explore the effect of platelet-activating factor on long-term potentiation in the CA1 region of rat hippocampal slices.

METHODSWe recorded the excitatory postsynaptic potentials (EPSPs) and investigated effect of long-term potentiation by PAF in rat hippocampus in vitro.

RESULTSLow doses (1 micromol/L ) of PAF could induce LTP, while higher doses (10-50 micromol/L) of PAF could inhibit induction of LTP. But it couldn't inhibit the LTP induced by subsequent high frequency stimulation and the EPSP of basal. GB of PAF receptor antagonists could prevent the LTP induced by low doses of PAF, and could't inhibit the LTP induced by HFS.

CONCLUSIONHigher doses of PAF is an HIV-1-induced neurotoxin, it may contribute to the HAD pathogenesis by inhibition of LTP.

Animals ; Electric Stimulation ; Hippocampus ; drug effects ; physiology ; Long-Term Potentiation ; Male ; Organ Culture Techniques ; Platelet Activating Factor ; administration & dosage ; pharmacology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the function of pancreatic islet beta cells, insulin resistance (IR) and features of TCM symptoms and syndromes in the non-diabetic first-grade relatives (ND1GR) of patients with type 2 diabetes mellitus (DM2).

METHODSA total of 68 ND1GR of DM2 patients were enrolled in the observed group and 45 healthy subjects with matched sex, age and body mass index (BMI) but without family history of DM were selected into the control group. Levels of fasting blood glucose (FBG), 2 hrs postprandial glucose (2hPG), fasting insulin (FINS) and 2 hrs postprandial insulin (2h INS) in all the subjects were measured to calculate and compare the IR and beta-cell function of the homeostatic model analog (HOMA-IR and HOMA-beta), and the insulin sensitive index (ISI). Moreover, the symptoms manifested in the ND1GR were also observed to analyze the features in them.

RESULTSFBG and FINS were obviously higher in the observed group than those in the control group (P < 0.01), while no significant difference was found in 2hPG or 2h INS (P > 0.05). HOMA-IR and HOMA-beta were significantly higher (P < 0.05) and ISI were significantly lower (P < 0.01) in the observed group than those in the control group. Compared with the control group, the main symptoms such as dark purplish tongue, listlessness, thready and thin pulse, lassitude in loin and legs in the observed group were seen more frequently. In the observed group syndrome of deficiency of Qi and Yin accounted for 51.47%, syndrome of deficiency of Yin for 30.88%, subjects with syndrome of blood stasis as the main accompanying syndrome accounts for 61.76%.

CONCLUSIONHigher beta cell secretion function and lower insulin sensitivity appear in ND1GR of DM2 patients, suggesting the existence of insulin resistance. The feature of TCM syndrome in them is characterized by deficiency of Qi and Yin with inner obstruction of blood stasis.

Adult ; Diabetes Mellitus, Type 2 ; genetics ; Diagnosis, Differential ; Female ; Glucose Tolerance Test ; Humans ; Insulin Resistance ; Insulin-Secreting Cells ; physiology ; Male ; Medicine, Chinese Traditional ; Middle Aged

OBJECTIVETo investigate the role of extracellular regulated kinase (ERK1/2) pathway in cisplatin-induced apoptosis in human ovarian carcinoma cells.

METHODSCisplatin-induced apoptosis were stained with DAPI and was assessed microscopically in human epithelial adenocarcinoma ovarian cell line SKOV3 cells. ERK activation was determined by Western blotting using an anti-phospho-ERK antibody to detect ERK activity. The effect of PD98059 on ERK activity induced by cisplatin was detected by MTT assay.

RESULTSMarked apoptosis of SKOV3 cells resulted from 48 hours treatment with 20 microg/mL cisplatin. Strong activation of ERK was led to by 15 microg/mL cisplatin. Dose response and time course of cisplatin induced apoptosis in SKOV3 cells. Cisplatin-induced ERK activation occurred at 12 hours and increased to highest induction at 24 hours by Western blotting. The effect of PD 98059 on ERK activity induced by cisplatin at the concentration of 100 micromol/L PD 98059. Statistically significant decreased in cell survival were observed with 100 micromol/L PD 98059 at 15 and 20 microg/mL cisplatin (P < 0.05).

CONCLUSIONSCisplatin activates the ERK signaling pathway in ovarian cancer cell line SKOV3. Inhibition of ERK activity enhances sensitivity to cisplatin cytotoxity in ovarian cancer cell line SKOV3. Evaluation of ERK activity could be useful in predicting which ovarian cancer will response most favorably to cisplatin therapy.

Adenocarcinoma ; enzymology ; pathology ; Antineoplastic Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Cell Survival ; drug effects ; Cisplatin ; pharmacology ; Enzyme Activation ; drug effects ; Female ; Flavonoids ; pharmacology ; Humans ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Ovarian Neoplasms ; enzymology ; pathology ; Signal Transduction

OBJECTIVETo define a correlation between different human papillomavirus (HPV) types and telomerase activity in cervical cancer.

METHODSTelomerase activity was detected by TRAP-PCR, and different HPV type was determined by PCR in 83 cervical cancer, 47 cervical intraepithelial neoplasia (CIN) and 10 normal cervix cases.

RESULTSWith regard to positive rates of telomerase and HPV 16/18: the results were cervical cancer > CIN > normal cervix, CIN III > CIN I, II; with regard to HPV 6/11 positive rate: the results showed CIN I, II > CIN III. Positive rates of telomerase cervical cancer and HPV were bearing on grading and staging, but they did not correlate with histologic subtypes. Positive rate of HPV 6/11 had nothing to do with grading, staging and histologic patterns. On expression strength of telomerase and HPV 16/18: the results showed cervical cancer > CIN, CIN III > CIN I, II. Regard to HPV 6/11'expression strength: the results showed CIN I, II > CIN III, CIN > cervical carcinoma.

CONCLUSIONHPV 16/18 infection seemed to have played an important role in carcinogenesis of cervical lesions by activation of telomerase.

Cervical Intraepithelial Neoplasia ; enzymology ; pathology ; virology ; Female ; Humans ; Neoplasm Staging ; Papillomaviridae ; isolation & purification ; Telomerase ; metabolism ; Uterine Cervical Neoplasms ; enzymology ; pathology ; virology

In order to find a method suitable for purifying large amount of CD34(+) cells, from 5 cases who accepted autologous peripheral blood stem cell transplantation, CD34(+) cells were collected and enriched by using Isolex 300i (Nexell). Phenotypes were detected by flow cytometry and the biological viability were assayed by the colony-forming experiments and cell expansion experiment in vitro. The results showed that the number of mononuclear cells first collected was about (3.5 - 6.0) x 10(10) and (0.55 - 1.2)% of cells were CD34 positive. The number of positive production was about (2.0 - 3.0) x 10(8); the CD34(+) cells purity was (75 - 85)% and the yield was (40 - 65)%. The CD34(+) cells of positive production could expand up to 2 - 3 times when cultured with SCF + IL3 + FL + TPO + EPO in vitro. The results of colony-forming experiments demonstrated that the CD34(+) cells collected has enough colony-forming ability. All results showed the enriched CD34(+) cells with biological viability. In conclusion, the CD34(+) immunomagnetic isolation apparatus Isolex300i is suitable to clinical application for a large amount of CD34(+) cell enrichment.
Antigens, CD34 ; immunology ; Colony-Forming Units Assay ; Flow Cytometry ; Hematopoietic Stem Cells ; cytology ; immunology ; Humans ; Immunomagnetic Separation ; instrumentation ; methods ; standards ; Neoplasms ; blood

To develop a Huntington’s disease(HD) cell model in vitro to screen drugs targeting the aggregation of polyQ,different length of CAG repeat fragments were amplified by random primer PCR, identified by DNA sequencing and were fused to the N-terminus of CAT in the pCAR system respectively which had been constructed and identified before. Recombinant plasmids were transformed into and induced to express in the host E.coli. SDS-PAGE and chloramphenicol resistance test were done to determine the solubility of the polyQ and chloramphenicol resistance levels of the fusions. With different length of CAG repeat fragments cloned and expressed in the CAT-fusion protein reporting system, it is found that when the length of the fragments increased over 40, their encoding polyQ expressed as insoluble protein and chloramphenicol resistance levels are lower, while under 40, the polyQ expressed as soluble ones and chloramphenicol resistance levels are higher. A in vitro HD model that could minimize the pathological process of the HD thus has been developed. With which by measure the recombinant bacteria’s resistance to chloramphenicol, the polyQ’ solubility and folding state in vitro by quality and quantity could be determined. Thus this model can be used to screen drugs or bioactivity materials that can inhibit aggregation of the polyQ, which thereby shedding new light on the prevent, diagnosis and therapy of HD.

Objective To study the genotypes of 102 strains of methicillin-resistant Staphylococcus aureus(MRSA)collected consecutively in 2002 in our hospital Method Multiplex PCR was used to genotype Staphylococcal chromosomal cassette mec(SCCmec)element and its variants.Results Among 102 strains of MRSA,the genotypes were as follows:SCCmec-Ⅲ(94 strains),SCCmec-ⅢA(4 strains), SCCmec-Ⅳ(2 stains),SCCmec-Ⅰ(2 stains).Conclusion The predominant genotype of MRSA circulating in this hospital in 2002 was SCCmec-Ⅲ by multiplex PCR.

OBJECTIVETo investigate the effects of Baisong tablet on the behaviors and CRHmRNA expression in the chronic stress rats.

METHODRats were exposed to different ways of chronic stress. Body weight and behaviors were investigated during the whole procedure, the CRHmRNA expressions in the hypothalamus and prefrontal cortex were semiquantified by the RT-PCR method.

RESULTIn comparision with the normal group, rats exposed to chronic stress showed decreased body weight and a significant reduction of consumption of sucrose solution, and the duration of immobility during the forced swimming test was increased significantly. The chronic stress rats was in depression of behavior. CRHmRNA expression in the brain of the chronic stress rats was upregulated significantly, while it was downregulated in the groups of Baisong tablet and the group of fluoxetin.

CONCLUSIONBaisong tablet has the effect of antidepressant, and it may be related to the effect of the downregulated CRHmRNA expression in brain.

Animals ; Behavior, Animal ; drug effects ; Body Weight ; drug effects ; Corticotropin-Releasing Hormone ; biosynthesis ; genetics ; Depression ; metabolism ; physiopathology ; Drug Combinations ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Female ; Frontal Lobe ; metabolism ; Hippocampus ; metabolism ; Male ; Plants, Medicinal ; chemistry ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Stress, Psychological ; metabolism ; physiopathology ; Tablets ; Tribulus ; chemistry