Objective: To reduce the risk of 3'-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU, WD) was designed to amplify a fragment of HBV DNA P gene by PCR, Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3'-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2. MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3'-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70. 4%(19/27) and 85. 2% (23/27) respectively (P

To reduce the risk of 3′-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU,WD) was designed to amplify a fragment of HBV DNA P gene by PCR. Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3′-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2, MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3′-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70.4%(19/27) and 85.2%(23/27) respectively (P<0.05). (2) The sequencing analysis of the PCR products suggested that the 3′-terminal mismatch of primers caused false negative in the PCR detection. Conclusion: When amplifying the variable region of DNA, the false negative result can be avoided by using 3′-terminus shifted degeneracy primers.

Aim To study the protection and possible mechanism of 5-hydroxy-1 H-indazole against 1-methyl-4-phenylpyridinium iodide ( MPP+)-induced SH-SY5 Y cell apoptosis. Methods An apoptotic model was es-tablished in human neuroblastoma SH-SY5 Y by MPP+in vitro. MTT analysis was used to evaluate the protec-tive effect of 5-hydroxy-1H-indazole. Immunochemistry and Hoechst33258 nuclear staining were used to ob-serve the neuroprotection and anti-apoptosis of 5-hy-droxy-1H-indazole. Western blot was used to detect the levels of P-tau ( Ser396 ) closely related to neuronal apoptosis and its upstream kinases:P-GSK-3β and CDK5 . Results MPP+ induced activation of GSK-3β, increase of activity of CDK5 , tau hyperphosphory-lation and neuronal cell apoptosis. However,5-hydrox-y-1 H-indazole reduced the activities of GSK-3β and CDK5,then decreased the level of tau hyperphosphory-lation and inhibited MPP+-induced SH-SY5 Y cells ap-optosis. Conclusions 5-hydroxy-1H-indazole could attenuate MPP+-induced SH-SY5 Y neuronal cell apop-tosis. Possible mechanism is that 5-hydroxy-1H-in-dazole inhibits GSK-3βand CDK5 two signal transduc-tion pathways to lower the level of tau phosphorylation, then plays a role of neuroprotection.

ObjectiveTo observe the effect of Buzhongyiqi decoction on post stroke fatigue.Methods60 stroke patients with fatigue were given Buzhongyiqi decoction for 4 weeks continuously. Chinese New Drug Clinical Guidelines was to evaluate the effect for patients according to improvement of symptoms.ResultsIn 60 cases, markedly effective in 47 cases (78.33%), effective in 10 patients (16.67%), invalid in 3 cases (5%). Symptoms of malaise, anorexia, muscle soreness were significantly improved.ConclusionBuzhongyiqi decoction can improve post stroke fatigue of stroke patients.

Objective To investigate the predictive value of clinical and radiographic features in fungal pathogen identification in immunocompromised patients with pulmonary invasive fungal infection (IFI).Methods All consecutive immunocompromised adult patients with pulmonary IFI in respiratory intensive care unit (ICU)in the First Affiliated Hospital of Xinjiang Medical University were recruited during a 2 year period.All patients met the 2008 European Organization for Research and Treatment of Cancer and Mycoses Study Group (EORTC /MSG) criteria were studied for proved or probable IFI responding to antifungal agents.The data of demographic,clinical and radiographic features,as well as serological test results of the patients were collected.Differences in the clinical and radiographic features of pulmonary IFIs caused by yeasts and molds were compared by χ2 test.A logistic regression model was used to perform discriminant analysis,and the effect of discrimination was assessed for accuracy.Results The study included 143 patients with a probable diagnosis of IFI who had the following risk factors:diabetes mellitus (43.4%),chronic lung disease (32.2%),broad-spectrum antibiotics administration (≥14 days;35.7%),malignancy (23.1%),corticosteroid therapy (≥14 days;23.1%),chronic renal failure and renal replacement therapy (16.1%),and immunological disease (10.5%).Frequent broad-spectrum antibiotics administration was associated with yeast infection (P <0.05 ),while mold infection was associated with chronic lung disease (P <0.05 ) .Yeast was more often isolated from patients with concurrent bacterial infection and on mechanical ventilation (P <0.05 ) . Thoracic high-resolution computed tomography (HRCT)showed the following images:bronchial pneumonia/pulmonary consolidation (53.1%),massive shadowing (29.4%),small nodules (24.5%),large nodules (18.9%),pleural effusion (18.9%),halo sign (14%),and cavity (9.8%).Imaging showed that mold was more common than yeast in patients with pleural and pericardial effusions (P <0.05).Logistic regression modeling showed that broad-spectrum antibiotics administration,prolonged mechanical ventilation,and pleural and pericardial effusions were statistically significant in fungal identification (P <0.05 ),with a predictive accuracy of 77.6%.Conclusions For immunocompromised patients with pulmonary IFI,most of the risk factors ,the main clinical and chest HRCT features did not help to predict the type of fungal pathogen,and yeast but not cryptococcus may be accompanied or colonized.

Objective To evaluate the effect of high-frequency chest well oscillation expectoration system on ventilator-associated pneumonia (VAP) and time for withdrawing mechanical ventilation (MV) system in ICU patients with invasive mechanical ventilation (IMV). Methods 100 ICU patients with IMV were divided into observationgroup (n = 50) and control group (n = 50). The high-frequency chest well oscillation expectoration was used in the former group and the mechanical vibration expectoration was used in the latter. The two groups were compared in terms of amount of sputum, vital signs (heart rate, breathing, systolic blood pressure and blood oxygen saturation), time for withdrawing MV system and VAP rate. Results On days 1, 2, 3, 4 and 5, the amount of sputum in the observationgroup was (33.5 ± 4.2)mL/d, (41.1 ± 3.0)mL/d, (38.2± 3 .5) mL/d, (34.8 ± 2.5) mL/d and (31.1 ± 2.1) mL/d, and those of the control group respectively was (27.4 ± 3.1) mL/d, (30.3 ± 3.6) mL/d, (28.1 ± 2.2) mL/d, (25.7±1.8)mL/d and (20.8 ± 1.7)mL/d. The differences between the two groups were statistically significant (P < 0.05). After sputum expectoration, the blood oxygen saturation of the observationgroup was significantly higher than the control group [(97.5 ± 0.9) vs. (95.2 ± 1.0)] (P <0.05), but there were no statistical differences in heart rate, breathing and systolic blood pressure (P > 0.05). The time for withdrawing MV system in the observationgroup and the control group respectively was (5.8 ± 2.2)d and (9.5 ± 1.8)d, (P < 0.05). The rates of VAP in the observationgroup and the control group respectively was 30.0% (15/50) and 52.0% (26/50), with significant difference between them (P < 0.05). Conclusion The high-frequency chest well oscillation expectoration for ICU patients with invasive mechanical ventilation can promote sputum expectoration , improve blood oxygen saturation , shorten the time for withdrawing the ventilator, and prevent the incidence of VAP.

Objective: To investigate the in vitro antibacterial activity of triclosan combined with different antibacterial agents against triclosan-resistant multidrug-resistant Acinetobacter baumannii (A.baumannii). Methods: A total of 626 A. baumannii strains were collected from the First Affiliated Hospital of Wenzhou Medical University from 2016 to 2017. The sensitivity of these A. baumannii strains to common antibiotics was detected by VITEK 2-compact automatical microbiological analyzer and the minimum inhibitory concentrations (MIC) of triclosan were detected by agar dilution method. Checkerboard method was used to detect the changes in MIC values of triclosan against 16 triclosan-resistant multidrug-resistant A. baumannii strains after it was used in combination with four external ointments, including gentamicin, erythromycin, chloramphenicol and kanamycin, and three common antibiotics of imipenem, meropenem and ciprofloxacin, respectively. Fractional inhibitory concentration index (FICI) was used to evaluate the joint bacteriostatic effects. Results: Among the 626 A. baumannii strains, 17 were resistant to triclosan with a drug resistance rate of 2.7% (17/626). These triclosan-resistant strains had high MIC values for ciprofloxacin, imipenem, ceftazidime and other commonly used clinical antibiotic and most of them were multidrug-resistant. After triclosan was used in combination with seven different antibacterial drugs, the MIC values of all drugs decreased to various degrees compared with those when they were used alone. Triclosan in combination with gentamicin, chloramphenicol and ciprofloxacin showed synergistic effects on 62.5%, 56.25% and 62.5% of the 16 strains and additive effects on 37.5%, 43.75% and 37.5%, respectively. When it was used in combination with erythromycin, kanamycin, imipenem and meropenem, synergistic effects on 37.5%, 25%, 12.5% and 12.5%, additive effects on 37.5%, 56.25%, 62.5% and 62.5%, and indifferent effects on 25%, 18.75%, 25% and 25% of the strains were detected, respectively. No antagonistic effect was found between triclosan and any of the above antibiotics. Conclusions Triclosan combined with gentamicin, chloramphenicol and ciprofloxacin had better in vitro antibacterial effects against the triclosan-resistant multidrug-resistant A. baumannii strains in this study with synergistic and additive effects. Some indifferent effects were found between triclosan and kanamycin, erythromycin, imipenem and meropenem, but no antagonistic effects were detected.

We describe the diagnosis and treatment of a very premature female infant with gastroschisis complicated by tracheostenosis. The pregnant woman, whose fetus was diagnosed with gastroschisis by ultrasound at 22 weeks in a local hospital, was admitted to the Second Affiliated Hospital of Wenzhou Medical University at 28 +1 weeks with oligohydramnios. Ultrasound after admission confirmed the previous diagnosis. A live baby girl was born by vaginal breech delivery at 29 +1 weeks after spontaneous rupture of the membranes. Because of the unstable oxygen saturation, the neonate finally received Silo in the delivery room prior to the closure of abdominal fissure 7 d after birth, and during the placement difficult endotracheal intubation was evident. She was diagnosed with having congenital tracheal stenosis via chest CT scans with 3-dimensional reconstruction 3 weeks after birth and received transbronchoscopic balloon dilatation at 3 months after birth. During the 2-year follow-up, she grew well without any complications.

Objective:To investigate the predictive value of neutrophil to lymphocyte ratio (NLR) for hemorrhagic transformation (HT) in patients with acute ischemic stroke (AIS).Methods:Consecutive patients with AIS without performing intravenous thrombolysis and mechanical thrombectomy admitted to the Cerebrovascular Disease Treatment Center, the Affiliated Brain Hospital of Nanjing Medical University from December 2015 to December 2020 were enrolled. The clinical, imaging and laboratory examination data were collected. HT was defined as the first imaging examination of AIS patients without finding bleeding signs, but the imaging reexamination after hospitalization found intracranial hemorrhage. Multivariate logistic regression analysis was used to determine the independent correlation between NLR and HT. The receiver operating characteristic (ROC) curve was used to analyze the predictive value of NLR for HT. Results:A total of 805 patients with AIS were included. The median age was 67 years (interquartile range, 63-71 years), the median National Institutes of Health Stroke Scale (NIHSS) score was 4 (interquartile range, 2-9), the median NLR was 3.84 (interquartile range, 2.66-5.30). Seventy-ywo patients (8.9%) had HT. There were significant differences in age, baseline systolic blood pressure, baseline NIHSS score, time from onset to admission, time from onset to blood collection, time from onset to imaging reexamination, NLR, atrial fibrillation, history of previous stroke and transient ischemic attack and stroke etiology between the HT group and the non-HT group (all P<0.05). Multivariate logistic regression analysis showed that NLR was an independent risk factor for HT in patients with AIS after adjusting for confounding factors (odds ratio 1.355, 95% confidence interval 1.099-1.672; P=0.005). The ROC curve analysis showed that the area under the curve of NLR predicting HT was 0.852, and the optimal cut-off value was 4.75. Its sensitivity and specificity of predicting HT were 88.3% and 71.8% respectively. Conclusion:High NLR is an independent risk factor for HT in patients with AIS during hospitalization, and had better predictive value for HT risk.

Neuro-enhancement，by intervening on nerves for non-medical purposes，improves people’s physical，mental，and cognitive functions. While benefiting people，it also raises ethical risks of privacy，fairness，autonomy，and identity recognition between themselves and “artificial life”. Faced with these serious ethical risk challenges，it is urgent to propose countermeasures that respect and safeguard the basic rights of human beings，promote fair benefits with the principle of priority，standardize the information dissemination of neuro-enhancement，strengthen public education and training of ethical on neuro-enhancement technologies and advance responsible innovation，as well as carry out ethical education for neuroscience practitioners，with a view to promoting the healthy development of the field of neuroscience.