To clone and express 27 kDa cysteine protease (CP) gene of Spirometra erinacei plerocercoid,and analyze the biology characteristics,a total of RNA was extracted from the plerocercoids and reversely transcribed into cDNA.The 27kDaCP gene was amplified by PCR and cloned into pM-19T vector for sequencing.The accurate sequence was subcloned into the expression vector pET-28a (+).The recombinant plasmid was transformed into Transetta (DE3) and the expression protein induced by IPTG.The recombinant protein was purified by Ni2 + affinity chromatography,and analyzed by SDS-PAGE and Western blotting.The 27 kDa CP gene and its expression protein were predicted and analyzed by bioinformatics analysis tools such as NCBI and ExPASy.Results showed that the ORF length of 27 kDa CP gene sequence was 1 011 bp,and the removed signal peptide sequence was 954 bp with the submission number of ANA52569 in GenBank.The whole sequence of 27 kDa CP (Mr 35 669.9,pI 5.92) was 317 amino acids conferred from cDNA,which belongs to the Peptidase_C39_like superfamily.The pET-28a (+)-27kDa-CP was expressed under the induction of IPTG.Western blotting analysis showed that the purified protein reach expectancy,and had better response with positive serum of Spirometra erinacei plerocercoid infection.In conclusion,the 27 kDa CP gene of Spirometra erinacei plerocercoid is successfully cloned and expressed and knowing coded sequences and bioinformatic.

Objective:To investigate the episodic future thinking in patients with schizophrenia from three per-spectives including the specificity,emotional valence,and content.Methods:Totally 25 patients with schizophrenia who met the Diagnostic and Statistical Manual of Mental Disorder,Fourth Edition (DSM-Ⅳ)criteria and 25 healthy controls matched with gender,age,and years of education participated in this study.The Sentence Completion for E-vents in the Future Test (SCEFT)was used to test episodic future thinking.Results:Schizophrenia patients showed deficits in episodic future thinking[(0.13 ±0.10)vs.(0.31 ±0.12),P <0.01].They had problems in imaging the specific events even after controlling for working memory and verbal memory [F(1,54)=6.60,P <0.05].More-over,they generated less positive events compared with the healthy controls [(0.36 ±0.20)vs.(0.48 ±0.16),P <0.01].As to the content,schizophrenia patients generated more events about hospitalization [0 (0,0.64)vs.0,P <0.01],more events that were unclassifiable [0.36 (0.09,0.64)vs.0.09 (0,0.36),P <0.01],and less events re-lated with personal career [0 (0,0.27)vs.0.18 (0,0.45),P <0.01].Conclusion:It suggests that schizophrenia patients show deficits in episodic future thinking.They have problems in imaging the specific events,and they have more negative thinking.

This paper describes a practical process for a SGLT2 inhibitor dapagliflozin. The target product was synthesized from 1-chloro-2-( 4-ethoxybenzyl)-4-iodobenzene and 2, 3, 4, 6-tetra-O-acetyl-alpha-D-glucopyranosyl bromide by iodine-magnesium exchange, and coupling and acetyl removing reactions with the total yield of 50%. This practical process highlights fewer reaction steps, less waste and mild reaction conditions.

Objective:To compare the effectiveness between three methods for purifying the immunoglobulin of egg yolk(IgY) which are polyethylene glycol (PEG) method, chloroform extraction method and chloroform / PEG method, and to provide basis for obtaining the batch of IgY.Methods:The inactivated vaccine of Vibrio parahemolyticus (V. parahemolyticus) was prepared and the hens were immunized by multi-point intramuscular injection.The eggs were collected and the IgY was purified by PEG method, chloroform extraction method and chloroform/PEG method.The protein extraction rate, the IgY titer and the purity of the antibody which purified by different methods were detected.Furthermore, the operation process, cost and safety of the three methods were analyzed.Results:The protein contents of the extraction belonging three methods from high to low in turn were chloroform extraction method, chloroform/PEG method, and PEG method.There was no significant difference in the antibody titer between three methods, and the tiler of chloroform extraction method was slightly high.The purities of purified antibody from high to low in turn were PEG method, chloroform/PEG method and chloroform method.The PEG method had better security but relatively lower extraction efficiency and higher cost.The chloroform/PEG method had high extraction efficiency and good antibody purity.Conclusion:The PEG method is suitable for a small amount of extraction in the laboratory.The chloroform/PEG method is appropriate for extracting the high quality IgY in a batch as it has high extraction efficiency and good antibody purity.

Objective:To compare the clinical effects of rapid maxillary expansion between tooth-borne and hybrid-borne expanders.Methods:PubMed,Cochrane Library,EMbase,CBM,CNKI,VIP and WanFang Data were searched from the date of their establishment to October 31,2015,to find clinical trials about comparison of rapid maxillary expansion by tooth tissue-borne versus hybrid-borne expanders.The quality of the included studies was evaluated by 2 independent reviewers,and Meta-analysis was performed by using RevMan 5.3 software.Results:8 articles with 206 cases were included.The results of Meta-analysis showed that:there were significant differences between the 2 groups in the changes of the right first molar dental inclination [MD =-1.62,95 ％ CI (-3.18,-0.07)],inter first premolar width [SMD =-0.86,95％ CI(-1.47,0.25)] and molar alveolar inclination [SMD =-0.86,95％ CI(-1.10,-0.20)].There was no significant difference between the 2 groups in the changes of inter first molar ~dth [SMD =-0.08,95％ CI (-0.6l,0.45)],the left first molar dental inclination [MD =-1.12,95％ CI(-2.57,0.33)],maxillary width [SMD =-0.30,95％ CI(-1.08,0.47)].Conclusion:The effect between tooth-borne and hybrid-borne expanders in the expansion of maxillary dental arch is similar.However,tooth-borne expanders may cause greater inclination of first molar and alveolar process.

Objective To study the effect of electroacupuncture on the behavior of APP/PS1 transgenic mice of different age, and to explore the optimal intervention time of electroacupuncture in treating Alzheimer's disease (AD). Method APP/PS1 double-transgenic mice of 4 months, 6 months and 9 months old, 20 in each age group, were randomized into a model group and an electroacupuncture group, and ten C57BL/6 wild-type mice were taken as a control group; after 6-week electroacupuncture treatment, the Morris water maze was adopted for spatial memory and behavioral test, and the changes of behavior in each group were observed.Result Of the 5-month-old mice, there were no significant between-group differences (P>0.05), while the time factor (day) produced a statistical significance (P<0.01); of the 7-month-old mice, there were significant differences considering the different groups, time factor and interaction (P<0.01); of the 10-month-old mice, there were significant differences considering the different groups, time factor and interaction (P<0.05). The spatial probe test showed that there were significant between-group differences in comparing the platform crossings and swimming distance in platform quadrant in each age group (P<0.05).Conclusion Electroacupuncture can improve the learning and memory of APP/PS1 double-transgenic mice, the age of 6-7 months old is possibly the optimal intervention time of electroacupuncture for AD, but there still requires further mechanism studies.

This article aims to summarize the significance of the establishment of human milk banks, the status of human milk banks in mainland China, analyze the relevant factors that affect the development of human milk banks, and propose corresponding countermeasures to provide a reference for improving the status of human milk banks in mainland China and promoting sustainable development.

Objective To analyze the clinical features and prognosis of anti-leucine rich glioma inactivated protein 1 (LGI1) encephalitis.Methods Twelve encephalitis patients with anti-LGI1 antibodies were collected from the First Affiliated Hospital of Zhengzhou University from June 2015 to December 2016.The clinical manifestations,electroencephalogram,laboratory examination and imaging findings were summarized and the prognosis was observed.The modified Rankin Scale (mRS) was used for evaluation before and after treatment.Results The major clinical features included memory deficit (10/12),spatial disorientation (7/12),epilepsy with generalized tonic-clonic seizures (9/12),faciobrachial dystonic seizures (7/12),hyponatremia (5/12),mental and behavioral abnormalites (1/12),light sleep (1/12),increased sleep (3/12),aphasis (4/12),dysphagia,choking (2/12),headache (1/12),dizziness (2/12),fatigue (2/12),ataxia (2/12),bradycardia (3/12),urinary disorders (2/12),intestinal obstruction (1/12),diarrhea (1/12).Admission mRS score was found to be three in eight cases,four in four cases.The abnormal electroencephalogram was found in six cases,mainly manifested as focal or diffuse slow wave,some accompanied by epileptic wave.MRI scan of brain showed abnormal signals in four cases,mainly involved medial temporal lobe,hippocampus,basal ganglia,while one patient avoided MRI scan due to implantation of pacemaker.Two patients presented with pulmonary nodules,one case with positive thyroid antibody and increased rheumatoid factor.The follow-up after treatment showed no one died;mRS score was two in two cases,one in nine cases and zero in one case;the sequelae were memory deficit,increased sleep,faciobrachial dystonic seizures.Conclusions Anti-LGI1 encephalitis is a treatable disease,cardinal clinical features of which are seizures,cognitive disorders,hyponatremia.Immunotherapy can improve the symptoms of the disease significantly,and the prognosis is better comparatively.

Diabetic kidney disease （DKD） is one of the typical microvascular complications in patients with diabetes and a major cause of end-stage renal disease， with the pathogenesis remains to be elucidated. It may be associated with hemodynamic effects， genetic factors， kidney inflammatory injury， oxidative stress， autophagy dysregulation， metabolic disorders and so on. Because of its complex mechanism， there are no specific prevention and treatment measures in clinical practice. AMP-activated protein kinase （AMPK）/mammalian target of rapamycin （mTOR） signaling pathway is a classical pathway involved in the regulation of autophagy. This pathway can be activated for treating DKD. Recent studies have demonstrated that the active components in Chinese medicinal herbs play a role in the prevention and treatment of DKD by directly acting on targeted cells and autophagy targets， which has attracted extensive attention. Researchers have extensively studied the occurrence and development of DKD and the mechanism of drug intervention in DKD， and the results prove that AMPK/mTOR pathway plays a role in the development of this disease. The active components in Chinese medicinal herbs regulate the AMPK/mTOR signaling pathway to affect autophagy， alleviate oxidative stress， inflammation， and extracellular matrix aggregation， and promote the generation of autophagosomes， thus mitigating kidney injury. This paper mainly reviews the relationship between AMPK/mTOR signaling pathway， autophagy， and DKD and the mechanism of active components in Chinese medicinal herbs in mediating autophagy via the AMPK/mTOR pathway， aiming to provide a theoretical basis for the clinical prevention and treatment of DKD.