Objective To observe whether bone marrow mesenchymal stem cell transplantation can improve vasospastic rats sense and motor function.Methods Rats grouped with randomized number method as Control group,Subarachnoid hemorrhage group.Stem cell culture media group and Stem cell transplantation group.Subarachnoid hemorrhage model were made with tail artery blood twice injection,2 days after 2' nd injection.Bone marrow mesenchymal stem cell were transplanted to lateral cistern.Subarachnoid hemorrhage(SAH) group didn' t transplant stem cell.Stem cell culture media group injected DMEM media as DMEM group.Stem cell transplantation group injected 30μl Bone Marrow mesenchymal stem cell suspension,so called BMSCs group.Neurofunctional score and learning memory expression were detected with morris mazer and Neurofunctional Score Scale in each group.Results After transplantation for 7 d,functional score of Control,SAH,DMEM and Stem cell group were 3.95 ±2.51,7.20 ± 1.03,7.23 ± 1.79 and 5.81 ± 1.11 respectively.Compared with others groups,Stem cell group score was significantly decrease(P=0.017).After transplanting stem cell for 14 d,the mean spanning plate time in Control group,SAH group,DMEM group and Stem cell group were 7.38 ± 1.73,4.52 ± 0.90,5.11 ± 1.93 and 7.32 ± 2.16 respectively,SAH and DMEM group vs other 2 groups,there were clearly statistically differences (P =0.009),while between control group and stem cell group,there were no statistically differences (P =0.14).Conclusion SAH rat transplant stem cell can improve sense,motor and learning expression in certain level.

To obtain the tervalent fusion toxin gene (named FT),three toxin gene fragments from three species of Vibrio parahaemolyticus,Vibrio vulnificus and Vibrio mimicus were connected with the flexible linker (GGGGS) using overla Pextension PCR. The three toxin gene fragments respectively encode the mature proteins of the thermostable direct hemolysin (TDH) of V. parahaemolyticus,the cytotoxin (VVC) of V. vulnificus and the heat-labile hemolysin (VMH) of V. mimicus. The identity of FT nucleic acid sequence was 99.6% with the corresponding toxin gene fragments. The open reading frame of FT was 3225 bp,encoding 1074 amino acid residues with the predicted molecular weight (MW) of 120.4 kDa. Then,FT was subcloned into the expression vector pET-22b(+). The construction of recombinant expression vector pET-22b-FT was followed by transforming into E. coli BL21(DE3) for expression. The SDS-PAGE electrophoresis results indicated that the MW of the fusion toxin protein was matched to the predicted MW. After induction by 1 mmol/L IPTG at 37℃,the fusion toxin protein was effectively expressed in E. coli BL21(DE3) with the amount of 11.49% through thin layer chromatography scanning (TLCS) analysis. Cavia cobaya was immunized using the purified cytorrhyctes to produce the anti-serum. Through the determination of the optimum working conditions,the sensitivity test,the specificity test,repeatability test and sample simulation test,the indirect ELISA method was established,which is a broad-spectrum,rapid and specific to detect various of food-poisoning Vibrio simultaneously.

Objective To investigate the effect of targeting vascular endothelial growth factor (VEGF) by microRNA-126 (miR-126) on neuronal damage in neonatal rats with hypoxic-ischemic encephalopathy (HIE). Methods Newborn 7 days old SD male rats were randomly divided into four group, sham operation group (group A), HIE group (group B), HIE+negative control group (group C), and HIE+miR-126 overexpression group (group D), eighteen in each group. After modeling, neurological deficit score and brain water content were measured. HE staining was used to observe the pathological changes of CAI area in hippocampus of brain in each group. Real-time PCR was used to detect the expression of miR-126 and VEGF. Immunohistochemistry was used to detect the expression of VEGF in CAI area in hippocampus of brain. Double luciferase target experiment was used to verify the targeting relationship between miR-126 and VEGF gene. Flow cytometry was used to detect neuron apoptosis in hippocampus. Western blotting was used to detect the expression of cleaved-Caspase-3 protein in brain tissue of rats in each group. Results There was no neurobehavioral damage in group A, the neurobehavioral score was 0, and the brain tissue was not damaged; the neurobehavioral scores in group B and group C were (2. 50±0. 55) and (2. 33±0. 82) respectively, and the brain tissue damage was obvious; the neurobehavioral score in group D was ( 1. 50 ±0. 55), and the damage of brain tissue was improved. Compared with the group A, the neurobehavioral score (P<0. 05) and brain water content of group B and group C increased significantly (P<0. 05); Compared with the group B, the neurobehavioral score (P<0. 05) and brain water content of group D (P<0. 05) decreased. Compared with the group A, the expression level of miR-126, VEGF mRNA and protein, neuron apoptosis rate and cleaved-Caspase-3 in brain tissue of group B and group C were all significantly lower (P<0. 05). Compared with the group B, the expression level of miR-126, VEGF mRNA and protein, neuron apoptosis rate and cleaved-Caspase-3 in hippocampus of group D were all significantly higher (P<0. 05). The result of luciferase reporter gene experiment showed that miR-126 and VEGF could be targetly binded. Conclusion Overexpression of miR-126 can reduce neuronal apoptosis in hippocampus of brain and improve the development of HIE. The mechanism may be related to the targeted inhibition of VEGF gene expression by miR-126.

Objective To investigate the clinical features of Guillain-Barre syndrome (GBS) and clarify the long-term prognosis of acute inflammatory demyelinating polyneuropathy (AIDP) and acute motor axonal neuropathy (AMAN). Methods We conducted an analysis of the clinical data of 50 patients with GBS treated in our hospital between 2003 and 2007. According to the eleetrophysiological criteria, the eases were classified into AIDP (n=24) and AMAN eases (n=22), with 4 eases of unknown classification. The patients unable to walk upon discharge were followed up for more than 6 months, and the clinical features and prognosis of the two groups were compared. Results The age, gender, cranial nerve involvement, Hughes grade at the initial neurological examination and at the time of symptom peak did not differ significantly between the patients with AIDP and AMAN (P>0.05), and most of the AMAN patients had a good recovery. The number of patients capable of walking at one month after the onset was comparable between the two groups (P>0.05). In AMAN group, the percentages of patients with slow recovery and those having rapid recovery were significantly higher than those in AIDP group (P<0.05). Conclusion The clinical recovery of AMAN patients can be either rapid or prolonged, and rigorous immunotherapy should be administered to achieve early recovery and ensure more favorable outcomes of the patients.

OBJECTIVETo establish an animal model for quantitative cigarette smoking and to determine the acute response of airways to cigarette smoke in guinea pigs.

METHODSThe device for inhaling quantitative cigarette smoking was made, which was double pass and single-direction with the minimum dead space. The changes of airway resistance(R(L))and dynamic lung compliance(Cdyn) in guinea pigs exposed to compound air consisting of 75% cigarette smoke and 25% oxygen were observed. Exudation of Evans blue in pulmonary vessels was also determined after consecutive inhalation of 60 ml smoke.

RESULTThe R(L) increased from the baseline of (0.21+/-0.05) cmH(2)O x ml(-1) x s to (0.37+/-0.13) cmH(2)O x ml(-1) x s after 10 consecutive breaths of cigarette smoke exposure(P<0.01). The Cdyn decreased to (61+/-19)% of baseline at the ninth to eleventh breaths (P<0.01). The exudations of Evans blue significantly increased in all measured parts of the airways such as lower trachea, main bronchi, proximal intrapulmonary airways and distal intrapulmonary airways (P<0.01).

CONCLUSIONThe model established in this study is useful for measuring the acute responses of airways induced by cigarette smoke in guinea pigs. Acute inhalation of cigarette smoke decreases dynamic lung compliance, increases airway resistance and vascular permeability of pulmonary vessels in guinea pigs.

Airway Resistance ; Animals ; Capillary Permeability ; Female ; Guinea Pigs ; Lung Compliance ; Male ; Models, Animal ; Smoking ; adverse effects

BACKGROUNDSeveral isoforms of p53 have been reported, which may have varying functions and expressions. This study aimed to analyze the expression patterns of p53 isoforms in renal cell carcinoma (RCC) at the mRNA and protein levels and their associations with clinical and pathologic factors to explore the mechanism of p53 isoforms' activity in RCC.

METHODSThe specimens of tumours (T) and clinically normal tissues (N) adjacent to them were collected from 41 patients with RCC. mRNA expression levels of p53 isoforms were detected using RT-PCR followed by nested PCR. Protein expression levels were detected using immunohistochemisty and Western blotting with the anti-p53 antibodies DO-1 and DO-12. The data were analyzed with clinicopathological features by chi(2) test or Fisher's exact test.

RESULTSp53 mRNA was expressed in all tumours and matched clinically normal tissue adjacent to the tumour. All six isoforms could be detected in tumour and normal tissues, with the exception of the Delta133p53beta isoform, which was not detected in the normal tissue. Of the six isoforms, p53beta mRNA was significantly overexpressed in tumour samples (P < 0.001), and correlated with tumour stage. Nested PCR results consistently indicated the presence of p53gamma (19T/22N), Delta133p53 (33T/26N), Delta133p53beta (2T/0N), and Delta133p53gamma (13T/9N). Immunohistochemical analysis showed that p53 was expressed only in tumour tissues and correlated with tumour stage and grade. The results of Western blotting analysis were consistent with these findings.

CONCLUSIONSAlthough all six isoforms are present in RCC, their function in tumour development or progression might be different. Our findings suggest that p53beta might play an important role in the formation of RCC and it might be used as a new predictor and therapeutic target for RCC.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Blotting, Western ; Carcinoma, Renal Cell ; genetics ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Humans ; Male ; Middle Aged ; Polymerase Chain Reaction ; Protein Isoforms ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Suppressor Protein p53 ; genetics ; metabolism ; Young Adult

Ocean is a unique and excellent resource that provides a diverse array of intriguing natural products. Marine natural products have demonstrated significant and extremely potent biological activities and have captured the attention of natural products chemists in the past few decades. It is increasingly recognized that a wealth of fascinating natural products and novel chemical entities will play a dominant role in the discovery of useful leads for the development of pharmaceutical agents and provide useful probes to lead to breakthroughs in a variety of life-science fields. This article focused on the research progress of chemistry of marine natural products in recent five years.
Alkaloids ; chemistry ; isolation & purification ; pharmacology ; Animals ; Anti-Infective Agents ; chemistry ; isolation & purification ; pharmacology ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Aquatic Organisms ; chemistry ; Biological Products ; chemistry ; isolation & purification ; pharmacology ; Humans ; Macrolides ; chemistry ; isolation & purification ; pharmacology ; Marine Biology ; Marine Toxins ; chemistry ; isolation & purification ; pharmacology ; Molecular Structure ; Peptides ; chemistry ; isolation & purification ; pharmacology ; Steroids ; chemistry ; isolation & purification ; pharmacology ; Terpenes ; chemistry ; isolation & purification ; pharmacology

OBJECTIVE: To explore the genetic etiology for a fetus with congenital orofacial cleft. METHODS: Single nucleotide polymorphism microarray (SNP array) was carried out on skin tissues sampled from the fetus following induced abortion for the detection of copy number variation (CNVs). Pathogenicity of the candidate gene was validated through experiment. RESULTS: SNP array revealed that the fetus has carried a hemizygous 9.23Mb deletion at Xq21.31-q22.1(91 063 807-100 293 555), which was inherited from its mother. The region contained 13 OMIM genes and 1 ncRNA coding gene(MIR548M). Inhibiting of the expression of the MIR548M gene in oral epithelial celllines has resulted in up-regulation of the expression of SUMO1 gene which was known to involve in the pathogenesis of orofacial cleft. CONCLUSION Dosage insufficiency of the MIR548M gene may underlie the etiology of orofacial cleft in this fetus.
Cleft Lip/genetics* ; Cleft Palate/genetics* ; DNA Copy Number Variations/genetics* ; Female ; Fetus ; Humans ; MicroRNAs/genetics* ; Polymorphism, Single Nucleotide ; Pregnancy ; SUMO-1 Protein

Objective: To investigate the prognostic value of dynamic monitoring of RUNX1-RUNX1T1 transcript in pediatric patients with t (8;21) acute myeloid leukemia (AML) . Methods: The clinical features and RUNX1-RUNX1T1 transcript levels of 55 pediatric t (8;21) AML patients, newly diagnosed from Jan. 2010 to Apr. 2016, were analyzed retrospectively. The relationship between the minimal residual disease (MRD) and prognosis was analysed by dynamic monitoring of RUNX1-RUNX1T1 transcript levels using real-time quantitative PCR (RQ-PCR) technology. Results: The RUNX1-RUNX1T1 transcript levels in bone marrow cells at diagnosis was not related to relapse. After one course of induction therapy, patients with a more than 2 Log reduction of RUNX1-RUNX1T1 transcript levels (>2 Log) had lower 5 years cumulative incidence of relapse (CIR) [ (24.3±8.4) % vs (52.6±9.7) %, χ²=9.046, P=0.003], relapse-free survival (RFS) [ (71.6±12.7) % vs (48.1±13.2) %, χ²=5.814, P=0.016], and better overall survival (OS) [ (76.9±12.5) % vs (48.9±14.7) %, χ²=6.346, P=0.012], compared to patients with a less than 2 Log reduction (a<2 Log) . Multivariate Cox survival analysis suggested that a>2 Log reduction in RUNX1-RUNX1T1 transcript levels after a course of induction therapy was an independent prognostic factor for RFS (HR=0.263, 95%CI 0.081-0.851, P=0.026) and OS (HR=0.214, 95% CI 0.057-0.808, P=0.023) . During consolidation therapy and follow-up period, molecular relapse of 16 cases and hematologic relapse of 13 cases were identified by continuous dynamic monitoring of RUNX1-RUNX1T1 transcript levels, with a median interval of 4.0 (1.5-5.8) months from the molecular relapse to hematologic relapse. 2 cases of molecular relapse who received timely allogeneic hematopoietic stem cell transplantation did not experience hematologic relapse. Conclusion Dynamic monitoring RUNX1-RUNX1T1 transcript levels by RQ-PCR technique can subdivide patients into relatively low and high risk group, early screen patients at high risk of relapse and provide a scientific basis for precision stratification and risk-adapted therapy for pediatric t (8;21) AML children.

Objective To investigate the source of the first human case of avian influenza A (H5N1) infection in Beijing. Methods Interviewing the relatives of the case and other key persons, collecting and detecting samples of related biological, epidemiological and environmental data of the case were conducted. Later, the infection source was thoroughly investigated. Results The case ever contacted a slaughtered duck 5 days prior to the onset of illness, and the duck was bought from a stall of a wet market in Yanjiao area of Hebei province. Ten environmental samples were collected in this stall and the neighboring stall of the market. Another 6 samples were tested positive for H5N1 virus by PCR method, with 5 virus strains isolated. The whole-genome sequencing indicated that the amino acid homology between the H5N1 virus strains from the environment and the virus isolated from the case reached 99.8%-100%. Conclusion From both epidemiological and virological evidence, it was proved that the first human case of avian influenza A (H5N1) infection in Beijing was infected by a duck that carrying H5N1 virus the case contacted 5 days proceeding the onset of illness.