[Effects of Mijiandao suppository on intestinal laxation in rats and its mechanisms].

Objective: To investigate the effects of Mijian Daotong Bowel Suppository (MJDs) on the compound diphenoxylate induced constipation model of male rats and its mechanisms. Methods: Sixty SD male rats were randomly divided into blank group, model group, positive group and MJDs group. The constipation model was established by using compound diphenoxylate gavage. The rats in blank group and model group were treated with saline by enema, the rats in positive group and MJDs group were given Kaisailu and honey decoction laxative suppository by enema, respectively, once a day for 10 days. The body weight, fecal water content, gastric emptying rate (GER) and carbon ink propulsion rate (CIPR) of rats were observed during modeling and administration. The effects of MJDs on the pathological changes of colon tissue in constipation rats were investigated by hematoxylin-eosin (HE) staining. The effect of MJDs on 5-hydroxytryptamine (5-HT) in the colon of constipation rats was investigated by ELISA kit. The effects of MJDs on the expressions of aquaporins 3 (AQP3) and aquaporins 4 (AQP4) in the colon of constipation rats were detected by immunohistochemistry. Results: After 10 days of administration, compared with the blank group, the body weight, fecal water content, carbon ink propulsion rate and colon 5-HT content in the model group were decreased significantly, while the expression levels of AQP3 and AQP4 in the colon were increased significantly (P＜0.05, P＜0.01). Compared with the model group, the fecal water content and colon 5-HT content in the positive group were increased significantly, and the expressions of AQP3 and AQP4 in the colon were decreased significantly. The body weight, fecal water content and colon 5-HT content in the MJDs group were increased significantly, and the expressions of AQP3 and AQP4 was decreased significantly (P＜0.05, P＜0.01). Compared with the positive group, the fecal water content of the MJDs group was decreased significantly, and the expressions of AQP3 and AQP4 in the colon of the MJDs group was decreased significantly (P＜0.05, P＜0.01). Gastric emptying rate was not statistically significant difference between the groups. Conclusion: MJDs has good therapeutic effects on constipation, and its mechanisms may be related to up-regulating the content of 5-HT in the colon and down-regulating the expressions of AQP3 and AQP4 in the colon.

Relapsing-Remitting Multiple Sclerosis Is Associated With Regional Brain Activity Deficits in Motor- and Cognitive-Related Brain Areas.

Objective: To identify the abnormal regional spontaneous brain activity associated with relapsing-remitting multiple sclerosis (RRMS) using fractional amplitude of low-frequency fluctuation (fALFF) analysis and their relationships with clinical features. Methods: A total of 26 RRMS (11 males, 15 females; age, 36.58 ± 10.82 years) and 27 status-matched healthy group (HGs; 12 males, 15 females; age, 35.85 ± 12.05 years) underwent an Expanded Disability Status Scale (EDSS) examination. fALFF was applied to evaluate the abnormal regional brain activity associated with RRMS. Pearson's correlation analysis was applied to calculate the correlations between the signal values of brain areas that exhibited abnormal fALFF values and clinical features. Receiver operating characteristic (ROC) curve was performed to evaluate the sensitivity and specificity of those altered brain areas to distinguish between RRMS and HGs. Results: Compared with HGs, RRMS exhibited higher fALFF in the right cerebellum posterior lobe, left orbitofrontal cortex, left dorsolateral prefrontal cortex, bilateral supplementary motor area, and right fusiform gyrus and lower fALFF values in the left hippocampus and right precuneus. ROC revealed that these areas showed two good and five fair AUC values (0.77 ± 0.03, 0.729~0.822). However, four combinations with more than five brain regions received the same best discriminatory power with a sensitivity of 96.3% and a specificity of 88.5%. EDSS revealed a negative correlation with supplementary motor area (r = -0.395, p = 0.046). Conclusions: RRMS is associated with abnormal regional brain activity deficits of motor- and cognitive-related areas. The fALFF parameter may serve as a potential biological marker to discriminate between the two groups.

Effectiveness and safety of oral Chinese patent medicines as adjuvant treatment for unstable angina pectoris on the national essential drugs list of China: a protocol for a systematic review and network meta-analysis.

INTRODUCTION: Achieving efficacious and safe treatments for unstable angina pectoris (UAP) is still a challenging clinical problem. The availability of different oral Chinese patent medicines frequently poses a practical challenge to clinicians, namely, which one to choose as first-line regimen for treatment. This study aims to examine the comparative effectiveness and safety of oral Chinese patent medicines for UAP on the national essential drugs list of China. METHODS AND ANALYSIS: We will conduct a network meta-analysis (NMA) of all randomised controlled trials to evaluate the use of oral Chinese patent medicines as adjuvant for the treatment of UAP. We will explore eight electronic databases from their inception to June 2018 and search for grey literature. Primary outcomes include mortality and the cardiovascular events. Secondary outcomes include: (1) symptom improvement; (2) ECG improvement; (3) frequency of acute angina attack; (4) duration of angina; (5) adverse effects. Two independent authors will screen titles and abstracts, review full texts, extract data, assess the risk of bias using the Cochrane risk of bias tool and assess the quality of evidence and strength of the recommendations using the Grading of Recommendations Assessment, Development and Evaluation (GRADE). If adequate data are available, NMA will be performed with Bayesian analysis methods. ETHICS AND DISSEMINATION: The NMA will help us to reduce the uncertainty of interventions and help clinicians to make optimal and more accurate therapeutic decisions for adults with UAP. Therefore, we will publish the findings of this study in a peer-reviewed journal. No ethics approval is necessary for this study based on the nature of its design. TRIAL REGISTRATION NUMBER: CRD42018092822.

Development and Validation of Prognostic Model for Patients with Acute Subdural Hematoma-Reliable Nomogram.

OBJECTIVE: By extracting clinical and computed tomography imaging data of patients with acute subdural hematoma (ASDH), factors that were significantly associated with poor prognosis were screened and a nomogram model was established and validated. METHODS: All patients with ASDH who underwent subdural hematoma removal and decompressive craniectomy from January 2014 to March 2018 in Qinghai Provincial People's Hospital were continuously collected. Finally, 124 patients were included in the study. According to the Glasgow Outcome Scale at 3 months after operation, patients were divided into a good prognosis group and a poor prognosis group. RESULTS: Univariate and binary logistic regression analysis were performed to screen out independent predictors that were significantly associated with poor prognosis of ASDH. On the basis of these factors, a nomogram model was established. CONCLUSIONS: The nomogram model had high accuracy for predicting poor prognosis in patients with ASDH, and it was easy to promote. In the future, large sample and multicenter prospective studies are necessary to complement and identify the results.

[Subtypes of muscarinic acetylcholine receptors involved in the persistent activity of layer V pyramidal neurons in the primary auditory cortex of young mice].

Cholinergic receptor activation and intracellular current injection lead to the persistent activity (PA), which may be involved in inducing neural plasticity. Our previous study showed that PA is closely related to the activation of muscarinic acetylcholine receptors (mAChRs) in pyramidal neurons of mouse primary auditory cortex (AI). However, the subtypes of mAChRs involved in PA remain unclear. Thus, using whole-cell patch-clamp recording and pharmacological methods, we investigated the role of different mAChR subtypes in inducing PA in AI layer V pyramidal neurons of young mice. The results showed that activation of mAChRs with intracellular depolarizing current induced PA in layer V pyramidal neurons. Blockade of M1, M2 or M3 subtypes prevented the PA, whereas M4 receptor antagonists did not affect the production of PA. The results suggest that the PA may be induced through a mechanism involving M1, M2 and M3 muscarinic receptors, but not M4 subtype.

A bibliometric analysis of clinical study literature of traditional Chinese medicine therapies for smoking cessation.

INTRODUCTION: Traditional Chinese medicine (TCM) is commonly used for smoking cessation in China. The aim of this study is to perform a comprehensive literature search to identify clinical studies on TCM therapies for smoking cessation. METHODS: Publications of randomized controlled trials, controlled clinical studies, cohort studies, case-control studies, case series and case reports, reviews and cross-sectional studies on smoking cessation using TCM therapies were retrieved from seven databases from their inception to February 2017. The following data were extracted and analyzed: study type, year of publication, language, country or region, journals, participants, intervention and comparison, and outcome. RESULTS: In total, 260 publications on TCM therapies for smoking cessation were identified from 1980 to 2016, including 52 randomized clinical trials, 7 controlled clinical studies, 1 cohort study, 110 case series, 18 case reports, 50 narrative reviews, 17 systematic reviews, and 5 cross-sectional studies. Of these, 68.5% (178) were published in Chinese and the remaining published in English. Mainland China (n=129, 49.6%) was the leading country in this field, followed by USA (n=27, 10.4%) and UK (n=25, 9.6%). A total of 36 645 participants from 40 countries with age ranging from 12 to 86 years were involved in 188 clinical studies (excluding reviews and cross-sectional studies). The most commonly reported therapies were auricular acupressure (25, 13.3%), body acupuncture (14, 7.4%), and body acupuncture plus auricular acupressure (14, 7.4%). Composite outcomes were most frequently reported (110, 58.5%). CONCLUSIONS: A substantial number of clinical studies have been conducted and published on TCM therapy for smoking cessation, mainly focusing on acupuncture stimulation techniques. The findings suggest that future research should pay more attention to acupuncture for smoking cessation.

Proteomic Analysis on Cercariae and Schistosomula in Reference to Potential Proteases Involved in Host Invasion of Schistosoma japonicum Larvae.

Schistosomiasis is a parasitic zoonosis posing great threat to human health. The infection is acquired by larval cercariae penetrating host skin and transforming into juveniles, schistosomula. Proteolytic enzymes secreted from the cercarial acetabular glands are known to aid to the skin penetration, but molecular mechanisms remain largely unclear. To profile the protein composition and identify potential invasive proteases, we developed a new method for simulating cercarial transformation and collecting schistosomula, and for the first time, we compared the proteomes of Schistosoma japonicum cercariae and schistosomula by using in-gel shotgun proteomic analysis. Totally, 1972 proteins were identified in association with ten main biological processes based on Gene Ontology analysis; 46 proteases were detected in cercariae, and among them, 25 proteases disappeared after penetrated. Notably, leishmanolysins and serine and cysteine proteases were found abundant but differentially expressed. Recombinant serine protease SjCE2b and cysteine protease SjCB2 were produced and used for validation of native proteins. Immunofluorescence and Western blotting assays detected SjCE2b and SjCB2 in cercariae but not in schistosomula, suggesting the two enzymes might be consumed upon skin migration. Our data comprehensively chart the proteomic changes during cercarial invasion, revealing the potential proteases involved, providing a platform for the development of molecular anti-infection strategy.

[In vitro whole-cell patch clamp recordings of neurons in subnuclei of mouse inferior colliculus].

The inferior colliculus (IC) is a pivot along the central auditory pathway. Using infrared visual whole-cell patch clamp recording technique, we investigated the electrophysiological properties of IC subnuclei neurons. Recordings were made from 88 neurons, including 21 neurons from the dorsal cortex of the IC (ICd), 43 neurons from the central nucleus of the IC (ICc) and 24 neurons from the external cortex of the IC (ICx). Based on the responses to positive current injection, three firing patterns, i.e., onset (6.8%, n = 6), adapting (39.8%, n = 35) and sustained (53.4%, n = 47) patterns, were identified. The hyperpolarization-activated inward current (Ih) could be recorded in half of the neurons (49/88). The sustained pattern occurred in more than half of ICd and ICc neurons (61.9% and 67.4%), while the adapting pattern occurred in majority of ICx neurons (75%). Action potential (AP) threshold and time constant also showed significant differences across neurons from the ICd, the ICc and the ICx. Our results indicate that IC neurons are different in electrophysiological properties across the subnuclei. The variance of the responses may be related to the distinct types of neurons as well as the received projections, which is implicated in the distinct roles of IC neurons in central auditory processing.

[Prokaryotic expression and function analysis of Schistosoma japonicum calpain].

OBJECTIVE: To clone and express recombinant calpain of Schistosoma japonicum (Sjcalpain), observe the distribution of Sjcalpain in S. japonicum cercariae and analyze its role in skin invasion. METHODS: The primers were designed according to the full-length sequence of calpain (GenBank accession No. AB016726). The genes encoding catalytic domain and Ca2+ binding domain of Sjcalpain were amplified by PCR, and the target fragments were subcloned into pET-28a. The recombinant proteins were expressed in E. coli BL21 (DE3) and purified by Ni-NTA resin. The rabbit polyclonal antibodies were prepared with the two purified recombinant proteins by immunizing New Zealand white rabbits. ELISA was used to detect the titer of rabbit antiserum. Immunolocalization was used to investigate the distribution of Sjcalpain in S. japonicum cercariae. Cercariae were incubated with specific inhibitor before infection of mice and the worm reduction rate was calculated. RESULTS: The recombinant expression vector Sjcalpain catalytic domain/pET28a and Sjcalpain Ca2+ binding domain/pET28a were constructed and the recombinant proteins were successfully expressed in E. coli BL21 (DE3) (about M(r) 43 000 and M(r) 39 000, respectively). The two target proteins were expressed as inclusion bodies. The purified target proteins were obtained through Ni-NTA affinity purification. ELISA result showed that the titer of prepared rabbit polyclonal antibodies was higher than 1 : 80 000. Immunolocalization study demonstrated that Sjcalpain protein was mainly expressed in the head of cercariae. Inhibition assays suggested that the average number of adult worms in calpain inhibitor-incubation group and control group was 19 and 23, respectively, with a worm reduction rate of 17.4%. CONCLUSION: Sjcalpain is mainly expressed in the head of S. japonicum cercariae. Inhibition of Sjcalpain could reduce the number of invading cercariae in infected mice, which suggest that Sjcalpain may play a role in skin invasion by cercariae.

[Cloning, expression and transcription specificity analysis of two tyrosinases from Schistosoma japonicum].

OBJECTIVE: To clone and express the recombinant proteins based on the whole open reading frame of two tyrosinases (tyrosinase 1 and tyrosinase 2) from Schistosoma japonicum, and study the transcription specificity of the two tyrosinases in different sex and developmental stages of S. japonicum. METHODS: The full-length of SjTYR1 and SjTYR2 were amplified with specific primers and subcloned into pSJ2. The recombinant plasmids were transformed into E. coli Rosetta Gami strains and induced with IPTG for expression. The recombinant proteins were purified by Ni-NTA agarose. The recombinant proteins SjTYR1 and SjTYR2 were used to produce the specific antibodies by immunizing the rabbits. The immunogenicity of the recombinant proteins SjTYR1 and SjTYR2 were detected by Western blotting using sera of recombinant proteins-immunized rabbits and S. japonicum-infected rabbit serum as the primary antibody, respectively. The reactivity of sera from recombinant proteins-immunized rabbits was analyzed by Western blotting against the native protein of S. japonicum worm. Total RNA was extracted from 14, 16, 18, 20, 22, 24, 26, and 28-day male and female worms. Transcription levels of the two tyrosinases in different sex and different stage were determined via RT-PCR method. RESULTS: The expression vector of SjTYR1/pSJ2 and SjTYR2/pSJ2 were constructed and the recombinant pro teins SjTYR1 and SjTYR2 were expressed in inclusion body in E. coli (about M(r) 55 000 and M(r) 56 800). The sera of S. japonicum-infected rabbits reacted positively with the purified recombinant protein SjTYR1, but not with recombinant protein SjTYR2. The native protein of S. japonicum worm could be recognized by sera of rSjTYR1-immunized rabbits (M(r) 100 000), but not by sera of rSjTYR2-immunized rabbits. Transcription levels of the two tyrosinases in male worms were nearly zero. In female worms, the transcription levels of the two tyrosinases increased sharply from the 24th day post-in- fection and reached maximum on the 28th day. CONCLUSION: The recombinant proteins of SjTYR1 and SjTYR2 show immunogenicity and immunoreactivity. SjTYR1 and SjTYR2 are both expressed specifically in female worms and the transcription levels increase in 24-28 days after infection.

Theoretical investigation on switchable second-order nonlinear optical (NLO) properties of novel cyclopentadienylcobalt linear [4]phenylene complexes.

As a kind of novel organometallic complexes, the cyclopentadienylcobalt (CpCo) linear [4]phenylene complexes (4 = number of benzene rings) display efficient switchable nonlinear optical (NLO) response when CpCo reversibly migrates along the linear [4]phenylene triggered by heating or lighting. In this paper, the second-order NLO properties for CpCo linear [4]phenylene complexes were calculated by using the density functional theory (DFT) methods with four functionals. All of the functionals yield the same order of ß tot values: 1<2<4<3. The effect of solvent on second-order NLO properties has been studied using polarized continuum model (PCM) in the tetrahydrofuran (THF) solution. The solvent leads to a slight enhancement of the NLO responses for the studied complexes relevant to their NLO responses in vacuo. The electronic absorption spectra were investigated by the TDDFT methods. The TDDFT calculations indicate that the maximum absorption peaks of complexes 2-4 in the near-infrared spectrum area show the bathochromic shift together with a decreasing intensity compared to complex 1. We have also found that the cobalt (Co) atom acts as a donor in all the organometallic complexes and the d → π* and π → π* charge transfer (CT) transitions contribute to the enhancement of second-order NLO response. Furthermore, two experimentally existing complexes 1 and 3 are found to have a large difference in ß tot values. It is our expectation that this difference may stimulate the search for a new type of switchable NLO material based on CpCo linear [4]phenylene complexes.