BACKGROUND: Mesenchymal stem cell transplantation for treatment of hypoxic-ischemic brain damage (HIBD) has obtained some outcomes in adult animals, but studies are few in neonatal animal models. Mesenchymal stem cells are commonly harvested from bone marrow. A few studies are on umbilical cord blood-mesenchymal stem cells (UCB-MSCs). OBJECTIVE: To investigate the feasibility and timeliness of UCB-MSC transplantation after injecting UCB-MSCs into neonatal rat models of HIBD. DESIGN, TIME AND SETTING: The complete randomized controlled animal experiment was performed at the Laboratory of Department of Neurology of First Hospital Affiliated to Nanchang University from October 2004 to July 2005. MATERIALS: A total of 38 healthy neonatal SD rats aged 7 days old were used to create rat models of HIBD. Three rats died. METHODS: Cord blood samples were collected after normal full-term delivery of 23-35 healthy pregnant women for culturing UCB-MSCs. MSCs were labeled with 4',6-diamidino-2-phenylindole 2hci (DAPI) in vitro before transplantation. Thirty-five rat models were divided into three groups. UCB-MSCs were injected into tail vein of twelve rat models in the early transplantation group two days after modeling. UCB-MSCs were injected into tail vein of twelve rat models in the late transplantation group one week after modeling. Same volume of saline was injected into eleven rats of the control group. Six rats from early transplantation and late transplantation groups each were respectively obtained at day 2 after transplantation and at week 2 after modeling. Three, four and four rats from control group were obtained respectively 2 days, 1 and 2 weeks after modeling, and sacrificed after anaesthesia. Ischemic brain tissues from the brain and hippocampal gyrum were sliced into frozen sections. MAIN OUTCOME MEASURES: Brain tissue pathomorphology was measured by Haematoxylin and Eosin Staining. Brain tissue DAPI-positive cells were detected with a fluorescence microscope. RESULTS: Brain edema at ischemic region, neural cell swelling and a decrease in cell number were tested in the control group. DAPI-positive UCB-MSCs were few in focal brain tissues, and swelling degree,extracellular space improvement and increased cell number were insignificant in the early transplantation group. One week after modeling, brain tissue extracellular space became small, cell number increased, and brain swelling reduced; A mass of DAPI-positive cells in rat focal brain migrated and diffused, without significant boundary in the late transplantation group. CONCLUSION: UCB-MSCs effectively traverse blood-brain barrier, and migrate, disperse and conform around focal brain tissues. A good outcome of transplantation is obtained at week 1 after HIBD.

Aims To establish several behavioral paradigms to characterize the psychotropic effects of hallucinogens which ze-brafish was utilized as a model animal, and then to investigate the effects of potent hallucinogenic drugs on these models. Methods With the video record and track system, the behavior was recorded and quantified automatically. In the experiments, the bottom dwelling test, social behavior and mirror test were performed to test the hallucinogenic effects of drugs. Metham-phetamine (METH, 2 mg·L-1) and ketamine (20 mg·L-1) were selected as experimental challenges. The 30 min pre-treat-ment time was chosen based on our prior experience in zebrafish models. Results Compared to the normal group, in dwelling test, acute exposure of zebrafish to METH and ketamine de-creased transitions significantly, and in mirror reflection test, the drug-treated fish changed the preference for mirror zone, and ex-hibited aggressive for their mirror images. The pretreatment of METH and ketamine significantly reduced the contact durations, and the ketamine inhibited the contact frequency each other, the results indicated that the social interaction of zebrafish was im-paired. Conclusion The results confirm high sensitivity of ze-brafish models to hallucinogenic compounds with complex behav-ioral and physiological effects.

Cas9 is a RNA-guided double stranded DNA nuclease that participates in the CRISPR/Cas9 system. Wide-type Cas9 directly silences the expression of target gene by gene splicing. The engineered dCas9 protein with the mutation at D10A and H840A lacks the Cas9' s endonuclease function but keeps its DNA binding activity. dCas9 can activate special genes by fusing with transcription activator. Meanwhile,it can inhibit the gene transcription by directly binding to the target gene and stop gene transcrip?tion. Combination of light sensitive structures and CRISPR can produce light-inducible CRISPR/Cas9 system for control of gene expres?sion. This system is able to activate or inhibit gene expression via the use of controlling blue light(470 nm). In this review,we mainly discuss the development of the light inducible CRISPR/Cas9 system as well as its application in the control of gene expression.

Hypoxia refers to the reduction in tissue oxygen supply or utilization. It occurs in various pathological symptoms like embolism,anthracemia,and chronic obstructive sleep. At high altitude, lower partial pressure of oxygen compromises the supply of adequate oxygen to the tissues and leads to many clinical syndromes,such as acute mountain sickness,high-altitude cerebral edema,and high-altitude pulmonary edema. Histamine H3 receptor, primarily as a presynaptic receptor, is widely expressed in the central and peripheral systems. Histamine,dopamine,acetylcholine and many other neurotransmitters are regulated by histamine H3 receptor. Studies have shown that histamine H3 receptor is involved in the hypoxic response of the respiratory network. In addition,histamine,espe?cially histamine H3 receptor,participates in the regulation of cerebral ischemia in the central nervous system. In this paper,we reviewed the structure and functions of histamine H3 receptor and explained its role in the regulation of hypoxia so as to evaluate the possibility of histamine H3 receptor as a drug target for the therapy of hypoxia-induced injuries.

OBJECTIVE To investigate the protective effect of sGC003,a novel agonist of soluble guanylate cyclase,on endothelin-1(ET-1)-induced cardiomyocyte hypertrophy. METHODS Cardiomy?ocytes were isolated from neonatal Sprague-Dawley rats using serial enzymatic digestion and then incubated with ET-1 10 nmol·L-1 in the absence or presence of sGC003 0.01,0.1 and 1.0μmol·L-1. Hyper?trophic responses including the cardiomyocyte area(Image-Pro Plus 6.0),the expression of atrial natri?uretic peptide gene(ANP)mRNA(RT-PCR method)and total protein content(BCA method)were detect?ed. RESULTS After 48 h stimulation with ET-1 10 nmol·L-1,the cardiomyocyte area increased by 80%(P<0.01),the total protein content increased by 120%(P<0.01) and the expression of ANP mRNA up-regulated by 140%(P<0.01). sGC003 0.01,0.1 and 1.0μmol · L-1 elicited antihypertrophic actions, including inhibition of ET-1-mediated increase in the cardiomyocyte area(P<0.01),raised total protein content(P<0.05)and upregulation of ANP mRNA(P<0.05). CONCLUSION sGC003 has protective,car?diomyocyte-selective antihypertrophic effects in vitro.

Schizophrenia, described as the worst disease affecting mankind, is a severe and disabling mental disorder. Schizophrenia is characterized by complicated symptoms and still lacks a diagnostic neuropathology, so developing schizophrenia animal models which have quantifiable measures tested in a similar fashion in both humans and animals will play a key role in new therapeutic approaches. According to the symptoms of cognitive impairment and emotional disorder, the N-methyl-d-aspartate (NMDA)-receptor antagonist MK-801 was applied to induce schizophrenia-like behavior in mice. Locomotor activity and prepulse inhibition (PPI) were selected as indices and the effect of clozapine was also investigated in this model. The results showed that compared with the normal group, MK-801-treated mice exhibited significantly increased locomotor activity and impaired PPI, and pre-exposure to clozapine could ameliorate the abnormality and make it back to normal level. These findings suggest that the model we established could be a useful tool for antipsychotic drug screening.

Objective To evaluate the diagnostic performance of a point-of-care testing for sensitive cardiac troponin Ⅰ (POCT-cTnI) in early diagnosis of chest pain patients who had a high pretest probability of acute myocardial infarction (AMI).Methods Total of 127 patients with new-onset chest pain at the emergency department were enrolled.Blood samples were drawn for the routine blood test,and determined POCT-cTnI and central laboratory testing for high sensitive cardiac troponin T (CLT-hscTnT) at admission,three and then at six hours after admission.All patients were divided into AMI group and non-AMI group according to the final diagnosis,which was adjudicated independently by two physicians who reviewed all available medical records for the 90-day follow-up period,and they were unaware of the results of the investigational assays of cardiac troponins.The receiver operating characteristic (ROC) curves were constructed to assess and compare the diagnostic performance of AMI of two cardiac troponin assays.The comparison of areas under the ROC curves (AUC) was performed by DeLong test,and the sensitivity,specificity,negative predictive values (NPV) and positive predictive values (PPV) for the target markers were calculated by applying a maker-specific cutoff value.Results The final diagnosis of AMI was made in 40 of 127 patients (31.5 ％).The diagnostic accuracy of the two assays oBtained at presentation,as quantified by AUC,was no statistically differences (AUC for POCT-cTnⅠ,0.901,95％ CI,0.901 to 0.947;and for CLT-hscTnT,0.907,95％ CI,0.842 to 0.951;Z =0.235,P =0.745).The AUC for POCT-cTnI at 3 hours after admission was significantly higher than that on admission (0.931 vs.0.858;Z =-2.038,P =0.042),while there was on further improvement at 6 hours after admission (0.931 vs.0.949;Z =-1.435,P =0.151).With use of POCT-cTnI (cutoff value 0.023 ng/mL,which was the 99th percentile upper reference limit) on adimission,the clinical sensitivity was 77.5％,and the specificity was 94.2％.A single sample of POCT-cTnI at 3 hours after admission improved the diagnostic accuracy,with a sensitivity of 96.4％,a specificity of 92.0％,and a NPV of 98.6％,a PPV of 81.8％.While,with use of CLT-hscTnT (cutoff value 0.014 ng/mL,was the 99th percentile upper reference limit) at 3 hours after admission,the NPV reached to 100％.Conclusions The use of a POCT-cTnI assay in chest pain patients can identify and exclude the AMI rapidly and exactly at three hours after admission,and the diagnostic performance is equivalent to CLT-hscTnT.

OBJECTIVE To investigate the effect of psychedelics on innate fear behavior of mice in Looming model(LM)and its neurobiological mechanism.METH-ODS ① Adult male C57BL/6J mice were randomly divid-ed into saline group,DOM group,psilocybin group and mescaline group.The drugs of the corresponding groups were given ip injecction 5 min in advance and LM were used to test the effect of them on freezing time in shelter of mice.② The mice were performing ip given DOM or psilocybin following 5-HT2A receptor antagonist M100907 ip 30 min later involved looming experiments.③Quan-tified the expression of EGR1 protein in mouse brains by immunofluorescence staining,then use ibotenic acid(IBO)damaged the mouse brain regions based on the result above and performed looming experiments.④ Specifically activate or inhibit CaMK Ⅱ,PV,VIP and SOM neurons of mice in saline or psilocybin groups respec-tively by chemical genetic methods and followed looming experiments.RESULTS ① In LM,the freezing time in shelter of mice in DOM,psilocybin and mescaline groups was significantly shorter compared to the saline group(P<0.05),and the dose-effect curves of above psyche-delics were U-shaped.② Compared with the vehicle + psilocybin/DOM groups,the freezing time in shelter of mice in M100907 + psilocybin/DOM groups was signifi-cantly prolonged(P<0.05),and there was no significant difference between the vehicle + saline group and the M100907 + psilocybin/DOM groups.③ Psilocybin signifi-cantly increased the expression of EGR1 protein in prelim-bic cortex(PrL)compared with saline,and the damage of PrL could effectively antagonized the effect of psilocybin shortening the freezing time in LM.④ Chemicalgenetic specific inhibition of CaMK Ⅱ,PV or VIP neurons in PrL could effectively antagonize the effect of psilocybin in LM,while chemicalgenetic specific activation of CaMK Ⅱneurons could significantly shorten the freezing time of saline-treated mice.CONCLUSION Psychedelics have capability to waken the innate fear behavior like freez-ing of mice in LM,and this effect is mediated by 5-HT2A receptor and CaMK Ⅱneuron in PrL.

Objective:To investigate the relationship between gallbladder carcinoma and gallbladder stones, and provide theoretical basis for the prevention and treatment of gallbladder carcinoma.Methods:A case-control study was used to retrospectively analyze the clinical data of 134 patients(study group) with gallbladder stones and gallbladder carcinoma treated in the Xinjiang Uygur Autonomous Region Corps Hospital of Chinese People′s Armed Police Forces from January 2010 to December 2012.Another 134 patients with gallbladder stones were selected as control group, and the clinical characteristics of the two groups were compared.Results:The average age of patients in the study group was (60.5±11.7)years, which was significantly older than that in the control group [(49.6±10.3)years], the difference was statistically significant ( t=7.916, P<0.05). The history of gallbladder stones in the study group and control group were (9.3±4.1)years and (4.6±2.5)years, respectively, and the difference between the two groups was statistically significant( t=11.682, P<0.01). The multiple stones, maximum stone diameter and maximum gallbladder wall thickness in the study group were 75 cases, (2.4±0.6)cm and (0.59±0.16)cm, respectively, which in the control group were 46 cases, (1.3±0.5)cm and (0.87±0.23)cm, respectively, the differences between the two groups were statistically significant(χ 2=3.978, t=6.217, 5.110, all P<0.05). The incidences of cholecystitis and jaundice in the study group were higher than those in the control group, and the differences were statistically significant(all P<0.05). Conclusion:Gallbladder stones are one of the causative factors of gallbladder carcinoma.Early diagnosis of gallbladder carcinoma is difficult.Patients with high-risk gallbladder stones who are old, have a long history of gallbladder stones, multiple stones, large stone diameters, and thick gallbladder walls should actively undergo surgical intervention.

ObjectiveThe functional model of six major components of Astragali Radix-Angelicae Sinensis Radix combination against the proliferation of vascular smooth muscle cells （VSMCs） was constructed by uniform design， the relationship between the compatibility of these six main components and the inhibition of VSMCs proliferation was analyzed， and the effect of the compatibility of these main components of Astragali Radix-Angelicae Sinensis Radix on the proliferation of VSMCs as well as the feasibility of uniform design test in the study of multi-component compatibility of Chinese medicines were discussed. MethodCell proliferation and toxicity assay kit （CCK-8） method was used to determine the inhibitory effect of the six components of Astragali Radix-Angelicae Sinensis Radix on platelet derived growth factor-BB （PDGF-BB）-induced VSMCs proliferation in rats and the half inhibitory concentration （IC₅₀） of each component were obtained. Six chemical components of Astragali Radix-Angelicae Sinensis Radix （formononetin， astragaloside Ⅰ， astragaloside Ⅳ， calycosin， ferulic acid and calycosin-7-O-β-D-glucoside） were taken as the independent variables X₁， X₂， X₃， X₄， X₅， X₆， respectively， and the cell proliferation inhibition rate as the dependent variable Y. U24*（24⁹） uniform design table and mathematical models （including linear and quadratic polynomial） were selected， the stepwise regression method was used to screen the variables， and the better equations were obtained in theory. The actually better relationship equations were screened by setting the different dose compatibility of these six components for verification tests. According to the purpose of the experiment， the selected regression equation was processed by data centralization， so as to compare the regression coefficients of the data centralization equation and analyze the correlation between components and efficacy. ResultThe six components of Astragali Radix-Angelicae Sinensis Radix could inhibit abnormal proliferation of VSMCs in a dose-dependent manner. IC₅₀ values of formononetin， astragaloside Ⅰ， astragaloside Ⅳ， calycosin， ferulic acid and calycosin-7-O-β-D-glucoside were 123.453， 113.184， 81.655， 141.159， 101.187， 151.016 mg·L^-1， respectively. Two superior models were selected by experimental verification， including equation 1 of Y₁=73.137 897-0.207 888X₁+0.000 508X₂²+0.000 347X₁X₆ and equation 2 of Y₂=70.038 433-0.236 665X₁+0.000 577X₂²+0.000 260X₁X₄+0.000 415X₁X₆+0.000 370X₃X₅. After data centralization， equation 2 was adjusted to Y=-1.480 260-0.146 281X₁+0.000 673X₂²+0.000 314X₁X₄+0.000 456X₁X₆+0.000 737X₃X₅， the regression coefficients showed that the astragaloside Ⅰ had a positive effect on the inhibition rate of cell proliferation， formononetin-calycosin， formononetin-calycosin-7-O-β-D-glucoside and astragaloside Ⅳ-ferulic acid had an interaction effect on the inhibition rate of cell proliferation， and the synergistic effect of astragaloside Ⅳ and ferulic acid was higher than other interaction items. ConclusionThe combination of six main components of Astragali Radix-Angelicae Sinensis Radix can inhibit the abnormal proliferation of VSMCs， and it is demonstrated that the uniform test design can be used to predict the correlation between the chemical composition of Chinese medicines with their pharmacological effects， which can provide an experimental basis for the rational use of uniform design and regression analysis to construct mathematical models to study the compatibility relationship of Chinese medicines.