Multilocus sequence typing (MLST) is a molecular genotyping method based on nucleotide sequencing. The procedure of this method characterizes isolates of bacterial species using the DNA sequencing of multiple housekeeping genes(usually seven). For each housekeeping gene, the different sequences present within a bacterial species are assigned as distinct alleles.For each isolate, the alleles at each of the loci define the allelic profile or sequence type (ST). MLST has the advantages of being robust (based on genetic data) and electronically portable to generate data that allow rapid and global comparisons between different laboratories. In this paper, the principle, method, data analysis, application, advantages and flaws of MLST are introduced.

Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS ( 1 μg/mL) and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay (ELISA), and the expression of miR-146a of cells was detected by using Real-Time PCR (TaqMan probe).Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student's t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge (P ＜ 0.01 ). The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells( P ＜0.01 ), and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.

Objective:To investigate the effect and mechanism of exosomes derived from human-induced pluripotent mesenchymal stem cells (iMSC-Exos) on alveolar macrophages (AM) pyroptosis.Methods:The exosomes in the culture supernatant of human-induced pluripotent mesenchymal stem cells (iMSC) were extracted by rotating ultrafiltration, and the extracted exosomes were identified by transmission electron microscopy, Western blotting and high-resolution adjustable resistance pulse. The rat alveolar macrophage cells (NR8383 cells) were cultured in vitro and the logarithmic growth phase cells were divided into three groups: the control group was added with an equal volume of phosphate buffered saline (PBS) in the AM supernatant; in LPS/ATP group AM cells were stimulated with 500 μg/L LPS for 23 hours and then 5 mmol/L ATP was added for 1 hour to induce pyrolysis; iMSC-Exos group was incubated with AM and 100 mg/L iMSC-Exos for 3 hours before giving LPS and ATP. The cytotoxic activity was detected by cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) analysis, the apoptosis and the expression of caspase-1 were observed by immunofluorescence, the levels of inflammatory factors interleukins (IL-1β and IL-18) released by AM were detected by enzyme linked immunosorbent assay (ELISA), the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the expression level of pyroptosis related protein gasdermin D (GSDMD) were detected by Western blotting. Results:The extracted exosomes were observed by transmission electron microscopy as round vesicles, expressing exosomal markers CD63 and CD9 showed by Western blotting, high-resolution adjustable resistance pulse showed the average diameter of the particles was 130 nm, and could be uptaken by AM. Compared with the control group, the cell activity decreased [(0.56±0.05)% vs. (1.06±0.07)%, P < 0.01], the release of necrotic substance LDH increased (U/L: 1 218.86±22.73 vs. 188.30±1.61, P < 0.01), the expression levels of inflammatory factors increased [IL-1β (ng/L): 958.91±32.78 vs. 194.63±5.14, IL-18 (ng/L): 870.89±21.86 vs. 288.85±24.48, both P < 0.01], and the apoptosis rate [(55.35±6.19)% vs. (12.01±1.32)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 41.06±3.65 vs. 2.80±0.54, P < 0.01) elevated in the AM after LPS/ATP stimulation, suggesting that LPS combined with ATP successfully induced alveolar pyroptosis. Compared with the LPS/ATP group, AM pretreated with iMSC-Exos showed increased cell viability [(0.81±0.05)% vs. (0.56±0.05)%, P < 0.01], decreased LDH secretion (U/L: 535.05±42.55 vs. 1 218.86±22.73, P < 0.01), decreased expression of inflammatory factors [IL-1β (ng/L): 381.82±19.50 vs. 958.91±32.78, IL-18 (ng/L): 533.77±31.54 vs. 870.89±21.86, both P < 0.01], and decreased apoptosis rate [(19.74±2.96)% vs. (55.35±6.19)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 12.16±1.31 vs. 41.06±3.65, P < 0.01). At the same time, the expression of NLRP3 inflammasome pathway [NLRP3 protein (NLRP3/β-actin): 0.62±0.06 vs. 1.89±0.11; cleaved caspase-1 protein (cleaved caspase-1/β-actin): 0.42±0.07 vs. 1.22±0.17, both P < 0.01] and pyrolysis-related protein was significantly inhibited [GSDMD protein (GSDMD/β-actin): 0.57±0.05 vs. 1.22±0.05, P < 0.01]. Conclusion:iMSC-Exos successfully reversed the AM pyroptosis and inflammatory factor expression induced by LPS/ATP, which may be due to the targeted inhibition of NLRP3 inflammasome pathway, suggesting that iMSC-Exos can exert anti-inflammatory effects by inhibiting the pyrolysis of AM.

Objective To learn the prevalence situation of dyslipidemia among adult residents in the southern coastal area, Wenzhou,Zhejiang.Methods Adult residents were chosen by using multi -stage sampling method from 11 counties in Wenzhou in 2013.Questionnaire survey,medical examination,and biochemical detection triglycerides were conducted among the residents.Chi -square test,and multiple logistic regression analysis was used to identify the risk factors related to dyslipidemia.Results The overall prevalence of dyslipidemia was 44.99%(the standardized prevalence rate was 42. 93%),and prevalence rate in women (43.91%)was higher than that in men (45.80%),but the urban(14.26%)and rural(13.83)areas prevalence was not significant.The awareness rate of dyslipidemia was only 21.73%.The prevalence of isolated low HDL -C,high LDL -C,hypercholesterolemia,hypertriglyceridemia,mixed hyperlipidemia was 13.95%, 10.45%,19.34%,12.98%,4.69%,respectively.Multivariate logistic regression analysis showed that gender,age, nationality,coronary heart disease,dyslipidemia,family history,aquatic products,milk,pickled products edible frequency,living and working pressure,body mass index,central obesity,hypertension,diabetes were related the prevalence of dyslipidemia,but the risk factors of different lipid fractions were different.Conclusion The prevalence of dyslipidemia has been in a high level among adult residents in Wenzhou,but the awareness rate of dyslipidemia was low. Measures should be Strengthened to prevent dyslipidemia .

AIM:To investigate the role of microRNA-132 (miR-132) on alveolar macrophage inflammation. METHODS: Rat alveolar macrophage cell line NR8383 was transfected with miR-132 mimic, mimic negative control ( NC) , miR-132 inhibitor, or inhibitor NC.The cells were divided into transfection group, transfection +lipopolysaccha-ride ( LPS) group, and transfection +LPS +acetylcholine ( ACh) group.The mRNA expression of acetylcholinesterase ( AChE) was detected by real-time PCR.The protein levels of AChE, signal transducer and activator of transcription 3 (STAT3) and phosphorylated STAT3 (p-STAT3) in the cells, and nuclear factor-κB (NF-κB) in the cytoplasm and nu-cleus were analyzed by Western blot.The activity of AChE in the culture supernatant was measured by AChE activity assay kit.The nuclear translocation of NF-κB was detected by immunofluorescence assay.RESULTS: Up-regulation or down-regulation of miR-132 had no effect on the mRNA expression of AChE.However, up-regulation of miR-132 decreased the protein level of AChE compared with mimic NC group (P<0.05).Transfection with miR-132 inhibitor increased the pro-tein expression of AChE compared with inhibitor NC group ( P<0.05 ) .In the alveolar macrophages treated with LPS+ACh, the inhibition of nuclear translocation of NF-κB p65 in miR-132 mimic group was more effective than that in mimic NC group ( P<0.05) .The inhibitory effect in miR-132 inhibitor group was weaker than that in inhibitor NC group ( P<0.05 ) .The inhibitory effect of miR-132 mimic on the protein levels of STAT3 and p-STAT3 was stronger than that of mimic NC (P<0.05).CONCLUSION:miR-132 in LPS-stimulated alveolar macrophages reinforced ACh-mediated anti-inflam-matory reaction by targeting AChE to suppress ACh hydrolyzation, which was related to the suppression of NF-κB and STAT3 activation.

Dunhuang medicine is a branch of Dunhuang studies, including the characteristics of Dunhuang ancient medical prescriptions and treatment technique.Dunhuang Dabupi Decoction has good efficacy in nourishing vitality and yin and promoting the production of body fluid. Professor WANG Dao-kun used the prescription to treat gastric stuffiness, gaining significant curative effect. This article summaried cases of using this prescription, to show the advantages of Dunhuang ancient medical prescription Dabupi Decoction for present use.

Spleen diarrhea is one of the common diarrhea diseases. Because it has long course, lingering and difficult to cure, and easy to occur, it is difficult to treat. Professor WANG Dao-kun believes that spleen-stomach yang deficiency, disorder of qi movement, and dampness and internal stagnation are the common etiology and pathogenesis of spleen diarrhea. The treatment should be warming middle Jiao and tonifying spleen, regulating qi, and promoting diarrhea. Professor WANG has prepared modified Wenzhong Zhixie Decoction for the treatment of spleen diuresis. This article expounded Professor WANG's experience in treating spleen diarrhea with Wenzhong Zhixie Decoction.

In view of that there is no report of west Nile virus infection cases in our country, evaluation the self-prepared anti-WNV-IgG diagnostic ELISA kit should be employed with the establishment of the serum sample panel collected from the entry personnel. All individuals of entry personnel were traveled from epidemic area of infectious west Nile disease. In our study, the serum samples were both detected by self-prepared anti-WNV-IgG diagnostic ELISA kit and the FDA certified kits ,which are FOCUS West Nile Virus IgG Dxselect and Panbio Dengue IgG Capture ELISA kits. The self-prepared kit and FDA certified kits were compared and assessed simultaneously. Furthermore, the specificity, repeatability and stability of the kits were also evaluated. The results indicated that no significant difference of detective rates (35. 6% for self-prepared kit vs. 32.5% for FOCUS kit, χ2 = 3. 05, P > 0.05) and good consistency (Kappa = 0.8372) between the self-prepared kit and FDA certified kits. Also, the positive coincidence rate, the negative coincidence rate and the total coincidence rate were calculated as 91.18%, 95.34% and 92.66%, respectively. The laboratory self-developed kit presented similar quality as the counterpart kits with FDA certificate. The development of our self-prepared anti-WNV-IgG diagnostic ELISA kit will provide technical support for the prevention and control of west Nile virus entry.
Endemic Diseases ; Enzyme-Linked Immunosorbent Assay ; Humans ; Immunoglobulin G ; blood ; Reagent Kits, Diagnostic ; West Nile Fever ; epidemiology ; West Nile virus ; immunology

OBJECTIVETo assess the long- term efficacy of surgical treatment of hepatic metastases from gastric adenocarcinoma.

METHODSClinical data of 938 cases with primary gastric cancer from January 1993 to October 2001 were analyzed retrospectively. Thirty- two cases received resection of hepatic metastases from gastric adenocarcinoma.

RESULTSMetachronous hepatic metastasis occurred in 24 cases(2.6% ),and 14 cases(58.3% ) underwent resection of hepatic metastases, while synchronous hepatic metastasis occurred in 90 cases(9.6% ) and 18 cases(20% ) underwent resection of hepatic metastases. The 1- 3- 5- year survival rates of hepatic resection for metachronous hepatic metastases were 73% 37% 25% respectively; while for synchronous hepatic metastases 68% 24% and 17% respectively. There were no significant differences between the two groups (P> 0.05).

CONCLUSIONSynchronous and metachronous hepatic metastases from gastric cancer should be treated by surgical resection to confirm a better prognosis.

Aged ; Female ; Hepatectomy ; Humans ; Liver Neoplasms ; secondary ; surgery ; Male ; Middle Aged ; Retrospective Studies ; Stomach Neoplasms ; pathology

OBJECTIVETo analyze the efficiency of cervical lymph node metastasis dissection and postoperative morbidity after selective three-field lymph node dissection (3FLND) for thoracic esophageal squamous cell carcinoma, and explore the proper selection conditions.

METHODSAccording to the conditions as follows: systemic evaluation, tumor T staging, tumor location, cervical CT and ultrasonography and the number of lymph nodes metastases, 85 patients with thoracic esophageal squamous cell carcinoma were selected and received 3FLND.

RESULTSIn the same period 45.5% (85/187) of the patients received 3FLND selectively based on the conditions. The rate of the cervical lymph nodes metastasis was 40.0% (34/85). The rate of the cervical positive lymph nodes of the upper, middle and lower thoracic esophageal carcinomas with enlarged lymph nodes suggested by cervical CT and ultrasonography was 68.4% (13/19), 41.7% (20/48) and 16.7% (1/6), respectively. Twelve patients with upper thoracic esophageal carcinoma with enlarged lymph nodes unrevealed by cervical CT and ultrasonography showed no histopathological lymph node metastasis. In the same period 17.1% (32/187) of the patients were selectively not undergone three-field lymph node dissection. The cervical lymph node metastasis rates in patients with upper and middle mediastinal lymph node metastasis were 79.3% (23/29) and 58.6% (17/29), significantly higher than 8.9% (5/56) and 7.1% (4/56) in the patients without upper and middle mediastinal lymph node metastasis (P<0.05). There was no in-hospital mortality in the group. The incidence of pulmonary complications and over-all postoperative morbidity was 24.7% and 42.4%, respectively.

CONCLUSIONSSelective 3FLND based on certain conditions can reduce the risk of postoperative morbidity and improve the efficiency of metastatic cervical lymph node dissection in thoracic esophageal squamous cell carcinoma. The thoracic tracheoesophageal groove positve lymph node indicated by CT scans should be one of selective conditions for 3FLND. The upper thoracic esophageal carcinoma should selectively receive 3FLND. The selection standards should be more strict for the lower thoracic esophageal carcinoma.

Carcinoma, Squamous Cell ; pathology ; surgery ; Esophageal Neoplasms ; pathology ; surgery ; Female ; Humans ; Lymph Node Excision ; methods ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Survival Rate ; Treatment Outcome