Objective To develop a sensitive, specific, rapid and easy method for detecting E. coli O157 : H7 and other Shiga toxin-producing E. coli in food and clinical specimens. Methods A circular probe and capture probe specific for Shiga toxin-2 (stx2) gene had been synthesized and was used for determining the sensitivity of ramification amplification method (RAM). Different serotypes which contained stx2 gene, including an E. coli O157 : H7, an E. coli O46 : H38, an E. coli O111 : NM, an E. coli O22 : H8 and E. coli ATCC23716 (stx2 gene negative) were used for determining the specificity. All strains were detected by RAM to determine whether they contained stx2 gene. Real-time RAM was further studied to detect stx2 gene. Results The lowest number targets detected by RAM assay was 10 copies of stx2, indicating that RAM assay was as sensitive as conventional PCR. The result of specificity showed that different serotypes of strains were all positive for stx2 gene, while nonpathogenic E. coli ATCC23716 was negative. Among 32 isolates, 28 STEC isolates containing stx2 gene were positive by RAM assay, while others were negative. The RAM results were 100% in concordance with that of PCR. The real-time RAM results also showed that as many as 10 bacteria can be detected and the time of appearance of detectable signal was depended on the target concentration. Conclusion RAM assay can offer an alternative method for PCR to detect E. coli O157 : H7 and STEC in all types of specimens because of its simplicity and isothermal amplification.

Humans ; Medical Waste Disposal ; Occupational Health

On the basis of studying processing method and quality control standard of Realgar,we did researches on the differences of toxicity and efficacy between new processing method(Washed by Acid)and traditional processing method(Grinding Without Water).The result showed that the efficacy was improved and the toxicity reaction Was lowered after Realgar being processed by new method.

Objective To explore the clinical diagnostic value of joint detection of serum platelet-derived growth factor BB (PDGF-BB), soluble vascular endothelial growth factor receptor -1 (sFlt-1) and urotensinⅡ( U-Ⅱ) in preeclampsia disease.Methods The cases of obstetric patients suffering from preeclampsia in the Third People′s Hospital of Liaocheng , Shandong Province between October 2012 and April 2014 were enrolled , including 96 cases of mild preeclampsia and 81 cases of severe preeclampsia.Totally 68 cases of normal pregnant women with similar age and gestational age were selected as control group.A case-control study was applied for the following investigations.The concentrations of serum PDGF-BB, sFlt-1 and U-Ⅱwere measured using ELISA.The diagnostic value of PDGF-BB, sFlt-1 and U-Ⅱalone or in combination for preeclampsia was analyzed and evaluated with receiver operating characteristic curve ( ROC) and Logistic regression analysis.Results Serum concentrations of PDGF-BB, sFlt-1 and U-Ⅱin mild preeclampsia group were (80.45 ±21.87)ng/L,(23.03 ±6.67)μg/L and(4.54 ± 1.02)ng/L, and those in severe preeclampsia group were (124.91 ±47.54)ng/L,(35.65 ±12.45)μg/L and(6.29 ±2.31) ng/L, while those in control group were (60.89 ±19.38) ng/L,(17.19 ±7.867)μg/L and ( 3.81 ±1.01 ) ng/L, respectively.The three parameters in mild preeclampsia group and severe preeclampsia group were significantly higher than those in control group ( P<0.01; F value was 79.43, 79.28 and 50.72 respectively ).In the same situation , these three indicators in severe preeclampsia group were significantly higher than those in mild group (P<0.01).Moreover, the AUC of serum PDGF-BB, sFlt-1 and U-Ⅱalone or in combination were 0.821, 0.786, 0.772 and 0.933, respectively.The differences between joint detection and three individual detections were statistically significant ( P<0.05 ) , as combined detection having a sensitivity of 95.7% and a specificity of 86.6%.Conclusion The combined detection of Serum PDGF-BB, sFlt-1 and U-Ⅱ had an important value in early assessment and treatment of preeclampsia.

Objective To examine the clinical factors associated with postoperative outcome in dementia combined with cerebral amyloid angiopathy-related intracerebral hemorrhage (CAA-ICH). Methods The clinical data of 25 surgical patients of dementia combined with CAA-ICH were retrospectively analyzed. The postoperative short-term effects and long-term effects were evaluated at the 30 d and 6 months after operation according to the modified Rankin score (MRS), and the influencing factors were analyzed. Results Among the 25 patients, eusemia was in 6 cases. Age ≥ 75 years, Glasgow coma score (GCS) ≤ 8 scores on admission, hypertension, postoperative anemia, pulmonary diseases, midline shift were the influencing factors of postoperative short-term effects (P < 0.05 or <0.01). Twenty-two patients were followed up for 6 months, and ensemia was in 9 cases. Age≥75 years, GCS ≤ 8 scores on admission, hypertension, postoperative anemia, hypoproteinemia and pulmonary diseases were the influencing factors of postoperative long-term effects (P<0.01 or<0.05). Conclusions Preoperative assessment is important for the patients of dementia combined with CAA-ICH. Age ≥ 75 years, GCS ≤ 8 socres on admission, hypertension, postoperative anemia, pulmonary diseases, midline shift are associated with poor short-term effects; age ≥ 75 years, GCS ≤ 8 scores on admission,hypertension, postoperative anemia, hypoproteinemia and pulmonary diseases are associated with poor long-term effects.

Objective To observe the influence of environment stimulation on learning and memory ability and hippocampal pathology of neonatal rats with hypoxic-ischemic brain damage (HIBD).Methods The models of HIBD SD rats were established by the method of Rice, and were divided randomly into three groups: enriched environment stimulation group (EE), impoverished environment stimulation group (IE), and standard environment stimulation group (SE). The sham-operation rats were served as control group. Different environment stimulation was administrated to the rats since day 1 after HIBD.On the day 28,Morris water maze was used to evaluate the learning and memory ability. HE staining and nissl stain were employed to observe the pathological change and the number of neurons in hippocampus of rats.Results The learning and memory ability of EE group was significantly higher than that of SE group (P0.05), and the ability of SE group was higher than that of IE group ( P0.05). The number of SE group was lower than that of Sham group (P

OBJECTIVE: To explore the methods of the larygeal recurrent nerve dissection in different and difficult thyroid surgery, so as to minimize damage and improve the safety of the operation. METHOD: The process and methods in different laryngeal recurrent nerve dissection about 52 hospitalized patients from 2010 to 2012 were retrospectively analyzed. These cases include large nodular goiter, nodular goiter behind the sternum or located in the lower pole of the thyroid gland, thyroid cancer, tumors of parathyroid gland, etc. We studied the conditions of lesions involving the laryngeal recurrent nerve and the defensive measures to protect the nerve. RESULT: The laryngeal recurrent nerve was dissected successfully in 50 cases, except 2 cases whose laryngeal recurrent nerve were violated by thyroid cancer. CONCLUSION When we dissect the laryngeal recurrent nerves in different and difficult thyroid, the glands and tumors were mostly needed to be freed and turned inward and forward. After that, the laryngeal recurrent nerves can be dissected successfully with the markers of tracheoesophageal groove, inferior thyroid artery and/or angle under the thyroid cartilage.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Recurrent Laryngeal Nerve ; surgery ; Retrospective Studies ; Thyroid Gland ; surgery

L9 (3(4)) orthogonal experiment was used to design the extraction technology of compound Clematidis Radix spray. Weight coefficients of active ingredients and dry extract rate were solved by information entropy. Support vector machine (SVM) was established and the model parameters were optimized through the genetic algorithm. Grid search algorithm was used for optimization of extraction technology of Clematidis Radix spray. The optimal extraction technology was to extract Clematidis Radix spray in water with 6 times the weight of herbal medicine for 3 times, with 2 h once. Bias of value between real and predicted by SVM was 1.23%. SVM was compared with traditional intuitive analysis of orthogonal design. It indicates that the new method used to optimize the extraction parameters of compound Clematidis Radix spray is more accurate and reliable.
Clematis ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Plant Roots ; chemistry ; Support Vector Machine ; Technology, Pharmaceutical

Objective To construct lentivirus containing CXCR4 gene and transfect MCF-7 cells , and obtain CXCR4 high-expressing MCF-7 cells. Methods CXCR4 gene was amplified by RT-PCR to construct CXCR4/pSin-EF2, which was transfected into HEK293T cells with psPAX2 and pMD2G vector for lentivirus packing. Packaged lentivirus was used to transfect human breast cancer cells MCF-7, with empty lentivirus as control. CXCR4 mRNA and protein expression levels were detected by RT-PCR and Western blot before and after transfection. And flow cytometry was used to detecte cell surface CXCR4 expression. Results The recombinant plasmid CXCR4/pSin-EF2 was constructed successfully,identified by double digestion and sequencing, and transfected into HEK293T cells to obtain high-titer lentivirus. RT-PCR and Western blot confirmed that the expression of CXCR4 in MCF-7 cells increased significantly after CXCR4 lentivirus transfection. Flow cytometry results showed that the CXCR4 positive rate increased from 26.78% to 99.29%, while there is no significant difference in CXCR4 expression between vector-transfected MCF-7 cells and non-transfected MCF-7 cells. Conclusion CXCR4 lentivirus and the breast cancer cell line with high and stable expression of CXCR4 (MCF-7CXCR4) were successfully constructed.

Objective The effects of candesartan,an angiotensin Ⅱ type 1 receptor blocker (ARB) were investigated on advanced glycation end-products accumulation and the receptor for AGE (RAGE) expression in type 2 diabetic KK/Ta mouse kidneys.MethodsKK/Ta mice(n=72)were random divided into three groups(n=24) and it was treated with candesartan [4 mg/(kg·d)] or vehicle from 6 or 12 to 28 weeks of age.BALB/c mice(n=24) treated with vehicle were used as controls.Body weight,blood pressure,blood glucose,urinary microalbumin,urinary creatinine and serum creatinine were measured every four weeks.At 28 weeks,renal expressions of carboxymethyllysine and RAGE were evaluated by immunohistochemistry and/or competitive RT-PCR.Results KK/Ta mice developed high body weight,high blood glucose,and high urinary microalbumin/creatinine ratio in KK/Ta mice at 28 weeks of age,and it was significantly higher than that of BALB/c mice [(427.49±89.37)mg/g vs (9.54±3.25)mg/g,P＜0.01 ].Protein and mRNA expressions of RAGE were upregulated in KK/Ta kidneys with increased immunostaining intensities of carboxymethyllysine.Candesartan treatment has markedly reduced urinary microalbumin/creatinine ratio [Early treatment group (32.18±9.41)mg/g,Late treatment group (53.20±7.26)mg/g,P＜0.01 ].Treatment with candesartan down-regulated the protein and mRNA expressions of RAGE and reduced the accumulation of carboxymethyllysine.There were no significant differences between the two treatment groups (from 6 or 12 weeks).ConclusionsThe results suggest that candesartan,an ARB,reduces advanced glycation end-products accumulation and subsequent albuminuria by down-regulating RAGE expression in type 2 diabetic KK/Ta mouse kidneys.