Objective To investigate the expression changes of the protein tyrosine phosphatase receptor T (PTPRT) in temporal lobe epileptic foci in the experimental animals and epileptic patients and the relationship between PTPRT and epileptogenesis.Methods After getting the epilepsy lobe tissue from the experimental and control groups,immunohistochemistry,immunofluorescence and Western blot analysis were used to assess the expression of PTPRT and its changes.Results In the temporal lobe tissue of intractable patients and control group,PTPRT was mainly expressed in the neurons.PTPRT was significantly increased in patients with intractable epilepsy (0.277 ± 0.048) than that in the control group(0.171 ±0.025 ; t =9.586,P ＜ 0.05).PTPRT was mainly expressed in the neurons in the temporal lobe brain tissue of the rats in the control group and experimental group.Compared with control group,the expression of PTPRT in the temporal lobe tissue was increased within 24 h post-seizure,and decreased 1 and 2 weeks post-seizure,then it was increased 1 and 2 months post-seizure (A ratio:control 0.443 ± 0.039,6 h 0.840±0.032,24 h 1.113 ±0.064,7 d 0.564 ±0.039,14 d 0.570 ±0.029,30 d 0.899 ±0.034,60 d 1.011 ± 0.074,F =256.427,P ＜ 0.05).Conclusions Through researches into the expression and function of PTPRT in the temporal lobe brain tissue of patients with intractable epilepsy and animal models,we presume that the PTPRT plays a role in the synapses reorganization and mossy fiber sprouting,and participates in the reconstruction of the neural network which leads to the intractable epilepsy.

OBJECTIVETo study the roles of stromal interaction molecule 2 (STIM2) and transient receptor potential canonical 3 (TRPC3) in extracellular Ca(2+)-sensing receptor (CaR)-induced extracellular Ca2+ influx and the production of nitric oxide (NO) in human umbilical vein endothelial cells (HUVEC).

METHODS(1) The interaction of STIM2 and TRPC3 was determined using the immunofluorescence technique. (2) The expressions of STIM2 and TRPC3 genes were silenced in HUVEC by transfection constructed STIM2 and TRPC3 RNA interference plasmids. The interference efficiency of STIM2, TRPC3 protein and mRNA levels were determined by Western blot and real time RT-PCR, respectively. (3) The second to fifth passage of HUVEC were divided into: STIM2-002 short hairpin RNA (STIM2-002 shRNA ) + spermine + Ca2+ group and TRPC3-004 short hairpin RNA (TRPC3-004 shRNA ) + spermine + Ca2+ group; control group (spermine + Ca2+ group) and vehicle+ spermine + Ca2+ group. The four groups of cells were incubated with CaR agonist spermine, the intracellular Ca2+ concentration ([Ca2+]i) was detected using the fluorescence Ca2+ indicator Fura-2/AM, and the production of NO was determined by DAF-FM (NO fluorescent probe) of each group in HUVEC.

RESULTS(1) Immunofluorescence technique results showed that STIM2 and TRPC3 proteinswere present in the cytoplasm of HUVEC. (2) The results of transfection constructed STIM2 and TRPC3 RNA interference plasmids demonstrated that shRNA targeted to the STIM2 and TRPC3 genes decreased STIM2 and TRPC3 mRNA levels by 88.2% and 74.0%, respectively (P < 0.05), simultaneously, the STIM2 and TRPC3 protein levels were decreased by 79.9% and 71.8%, respectively (P < 0.05). (3) Compared with spermine + Ca2+ group, the [Ca2+]i and the net NO fluorescence intensity of spermine + Ca(2+) + ShSTIM2-002 group, spermine + Ca(2+) + ShTRPC3-004 group and spermine + Ca2+ Vehicle group were not changed (P > 0.05).

CONCLUSIONSTIM2 and TRPC3 do not participate in CaR-mediated Ca2+ influx and NO production individually.

Calcium ; metabolism ; Cell Adhesion Molecules ; physiology ; Cells, Cultured ; Human Umbilical Vein Endothelial Cells ; physiology ; Humans ; Nitric Oxide ; metabolism ; Stromal Interaction Molecule 2 ; TRPC Cation Channels ; physiology

OBJECTIVETo discuss the diagnosis and treatment of primary hepatic carcinoid tumor (PHCT).

METHODSReport one case of huge PHCT treated in February 2004, and search the other 19 cases which were published from January 1994 to December 2006 in the Chinese biological and medical literature database. The clinical manifestation, pathological findings, diagnosis and treatment of these 20 PHCT patients were analyzed retrospectively.

RESULTSThe main symptoms were abdominal pain or discomfort (8 cases) and abdominal mass (7 cases), cases with typical carcinoid syndrome were rare (3 cases). Immunohistochemical staining was positive for neuron-specific enolase, chromogranin A and synaptophysin in most cases. Sixteen cases received operation, among which there were 13 removed completely, other 4 cases were treated by transcatheter arterial chemoembolization (TACE).

CONCLUSIONSThe definite diagnosis of PHCT depends on pathological and histochemical findings. Complete surgical resection is the best treatment for PHCT with favourable prognosis. TACE is also effective for nonoperative cases.

Antigens, CD34 ; analysis ; Carcinoid Tumor ; diagnosis ; metabolism ; therapy ; Chromogranin A ; analysis ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Liver Neoplasms ; diagnosis ; metabolism ; therapy ; Male ; Middle Aged

OBJECTIVETo evaluate the ability of a polycaprolactone/polylactic acid (PCL/PLA) membrane to inhibit epidural scar adhesion after laminectomy, and observe the responsive changes of the pain media in the spinal cord.

METHODSL(1), L(3) laminectomies were performed on 96 Wistar rats. The rats were divided into 3 groups: None-implant Control Group (NC), Autologous free fat graft group (AFFG) and PCL/PLA membrane group (PCL/PLAm). The rats were killed at 1, 3, 6, and 12 weeks postoperatively. Epidural scar formation and adhesion were observed grossly and histologically. Reverse transcription polymerase chain reaction (RT-PCR) were used to analyses the expression of Transforming growth factor beta (TGF-beta) in the epidural scar. Immunohistochemistry stain and RT-PCR were performed to evaluate the expression of the substance P and the c-fos gene in the relevant spinal cord, and the results were analyzed statistically.

RESULTSGross evaluation and histological evaluation showed that in the NC lamina defect site had much scar tissue and had wide and tight adhesions to the dura; in the AFFG, with the fat degrading gradually, the adhesions were increased; whereas in the PCL/PLAm group, there were slightly adhesions to the dura. RT-PCR showed that the expression of the TGF-beta was much less in the PCL/PLAm group than in the NC group. The insertion of the PCL/PLA membrane and the fat patch reduced the expression of the substance P and the c-fos gene in the spinal cord.

CONCLUSIONThe insertion of the PCL/PLA membrane reduces scar formation and separates fibrosis tissue from the dura, the results indicate that PCL/PLA membrane is an effective way of reducing peridural scar formation and preventing the failed back surgery syndrome.

Animals ; Biocompatible Materials ; Cicatrix ; prevention & control ; Female ; Lactic Acid ; Laminectomy ; adverse effects ; Membranes, Artificial ; Polyesters ; Polymers ; Postoperative Complications ; prevention & control ; Prosthesis Implantation ; Proto-Oncogene Proteins c-fos ; biosynthesis ; Rats ; Rats, Wistar ; Spinal Cord ; metabolism ; Spinal Diseases ; prevention & control ; Substance P ; biosynthesis ; Tissue Adhesions ; prevention & control

This study was purposed to investigate the effect of crocin on the proliferation in vitro and immune function of dendritic cells (DC) derived from the bone marrow of children with acute leukemia. The mononuclear cells were isolated from bone marrow of leukemia children by Ficoll-Hypaque. The experiment was divided into six groups: blank control group (A), crocin 1.25 mg/ml group (B), cytokines (rhGM-CSF 75 ng/ml+rhIL-4 75 ng/ml+rhTNF-α 50 ng/ml) group (C), cytokines+crocin 0.3125, 1.25 or 5.0 mg/ml groups (D, E, F). The numbers of DC were counted and the phenotypes of DC were determined by flow cytometry on the ninth day of culture. The DC of different groups were mixed with T cells just separated from peripheral blood of another children with acute lymphoblastic leukemia, and cultured with rhIL-2 200 U/ml for 5 d. The function of DC was detected by mixed lymphocyte reaction (MLR). The results indicated that the test groups and control group all obtained a certain amount of typical DC, but the DC numbers in test groups were all higher than those in control group (P < 0.01). Cultured for 9 days, the rates of CD1a(+), CD83(+), and HLA-DR(+) in group C, D, E, F were higher than group A (P < 0.01). There was no statistically significant difference between A and B groups (P > 0.05). MLR showed that with the increasing of DC, the stimulation index of T cells in group A and B was not rising (P > 0.05); the stimulated index of T cells in group C and E was significantly rising, there was statistically significant difference between them (P < 0.01). When the number of stimulated cells was the same, the stimulation index of T cell in group E was the highest (P < 0.01). It is concluded that the capability of DC proliferation promoted by crocin alone is lower than that of its combination with rhGM-CSF, rhIL-4 and rhTNF-α, but the crocin can synergically promote the maturity of DC cooperating with rhGM-CSF, rhIL-4 and rhTNF-α. The DC induced by crocin can particularly enhance the proliferation of T cells.
Bone Marrow Cells ; cytology ; drug effects ; Carotenoids ; pharmacology ; Cell Proliferation ; drug effects ; Child ; Dendritic Cells ; cytology ; drug effects ; Humans ; Leukemia ; pathology ; Lymphocyte Culture Test, Mixed ; T-Lymphocytes ; cytology ; Tumor Cells, Cultured

The aim of the study is to establish and evaluate a RNA isothermal transcription-mediated amplification and realtime detection assay (RIARD-MF) for the identification of Mycobacterium fortuitum in clinical isolates.RNA probes and specific primers of reverse transcription and amplification for T7 promoter were designed based on the sequence of M.fortuitum 16S rRNA.The isothermal successive cycles of amplification were performed for real-time detection by using T7 RNA polymerase at 42 ℃.Five non-mycobacterium strains,20 Mycobacterium strains and 259 clinical strains were detected by the established assay to evaluate the specificity and sensitivity,and the results were compared with those of PCR sequencing.In the test of 5 non-mycobacterium strains and 20 Mycobacterium strains,only M.fortuitum was positive,and the remaining 24 strains of bacteria were negative,which was consistent with PCR gene sequencing.The sensitivity and specificity of RIARD-MF reached 60 CFU/mL and 100％.In the test of 259 strains of clinical isolates,5 strains were identified to be M.fortuitum,the remaining 254 strains were not identified to be M.Fortuitum,which was also consistent with PCR gene sequencing.Both the specificity and sensitivity reached up to 100％ in the detection of clinical isolates.It suggested that the RIAR-DMF established in this study is a specific,sensitive,accurate and rapid method for the identification of M.Fortuitum and it may be hopeful for rapid identification of M.fortuitum in clinical isolates.

OBJECTIVE: To culture human gingival epithelia in vitro, and to construct the tissue engineered gingiva with the acellular dermal matrix (ADM). METHODS: Human gingival epithelia were isolated from the gingival tissue, and the cells were cultured and seeded onto the surface of ADM. After 7 days of submerged incubation, an air-liquid interface culture was performed for 7, 14, and 21 days. The complex constructed above was taken for histological examination. RESULTS: Human gingival epithelia could proliferate well on the surface of ADM, and form multilayer structure. But the superficial epithelium was partially keratinized. CONCLUSION Tissue engineered gingiva may be constructed with human gingival epithelia and ADM in vitro.
Acellular Dermis ; Cells, Cultured ; Connective Tissue ; Epithelial Cells ; cytology ; Gingiva ; cytology ; Humans ; Skin, Artificial ; Tissue Engineering ; methods

Since the research of molecular identification of Chinese Materia Medica (CMM) using DNA barcode is rapidly developing and popularizing, the principle of this method is approved to be listed in the Supplement of the Pharmacopoeia of the People's Republic of China. Based on the study on comprehensive samples, the DNA barcoding systems have been established to identify CMM, i.e. ITS2 as a core barcode and psbA-trnH as a complementary locus for identification of planta medica, and COI as a core barcode and ITS2 as a complementary locus for identification of animal medica. This article introduced the principle of molecular identification of CMM using DNA barcoding and its drafting instructions. Furthermore, its application perspective was discussed.
Animals ; China ; DNA ; genetics ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; classification ; isolation & purification ; Electron Transport Complex IV ; genetics ; Materia Medica ; classification ; isolation & purification ; Medicine, Chinese Traditional ; Plant Proteins ; genetics ; Plants, Medicinal

BACKGROUNDBacterial vaginosis (BV) is one of the most common infectious diseases among sexually active women and is associated with the increased acquisition of a variety of sexually transmitted diseases. This study aimed to compare the efficacy of a non-antibiotic sucrose gel against an antibiotic metronidazole gel for the treatment of BV.

METHODSA randomized, double-blinded, multi-center, parallel-group, placebo-controlled phase III clinical trial was conducted at eight hospitals in China. A total of 560 subjects with clinically diagnosed BV were randomly assigned into three groups for vaginally receiving sucrose, metronidazole, and placebo gels, respectively, twice daily for five consecutive days. The efficacy of therapeutic cure, defined as an achievement of both microbiologic cure (a Nugent score of 3 or less) and clinical cure (a resolution of the clinical findings from the baseline visit), was evaluated at the 1st and 2nd test-of-cure (TOC) visits at 7-10 and 21-35 days after the start of treatment, respectively.

RESULTSTherapeutic cure rates for sucrose, metronidazole, and placebo gel groups were 83.13%, 71.30% and 0.92%, at the 1st TOC, and 61.04%, 66.67% and 7.34%, at the 2nd TOC, respectively. While there was no significant difference between the sucrose and metronidazole gel groups at the 2nd TOC (P = 0.305), and sucrose gel was more effective than metronidazole gel at the 1st TOC (P = 0.009).

CONCLUSIONThese findings suggest that sucrose gel restores normal vaginal flora more rapidly than metronidazole gel and can be used as a novel treatment for BV.

Administration, Intravaginal ; Adolescent ; Adult ; Anti-Bacterial Agents ; administration & dosage ; therapeutic use ; Double-Blind Method ; Female ; Humans ; Metronidazole ; administration & dosage ; therapeutic use ; Middle Aged ; Sucrose ; administration & dosage ; therapeutic use ; Treatment Outcome ; Vaginosis, Bacterial ; drug therapy ; Young Adult

This study was to investigate the proliferative inhibition and apoptosis of human leukemia HL-60 cells induced by crocin and their possible mechanisms. The cell viability was tested by cell counting. The morphology of HL-60 cells was observed by fluorescence microscopy. The MTT assay was used to evaluate the inhibitory effect of crocin on the growth of HL-60 cells. Flow cytometry was used to measure the cell cycle. RT-PCR was used to detect bcl-2 and bax expression. The results indicated that the growth of HL-60 cells was inhibited remarkably in the dose and time dependent way. When the crocin concentration was higher than 5 mg/ml, the percentage of apoptotic HL-60 cells was not increased, on the contrary this percentage decreased, the cells manifested necrosis. Flow cytometry profiles revealed that cells were blocked in G₀/G₁ phase, the cell proliferation was inhibited obviously at 5 mg/ml. RT-PCR detection revealed that the expression of bcl-2 was down-regulated strikingly and bax was up-regulated. It is concluded that the crocin can inhibit the proliferation of HL-60 cells effectively, and block cells in G₀/G₁ phase. The mechanisms by which crocin induced apoptosis in HL-60 cells may be related to the inhibition of bcl-2 and activation of bax.
Apoptosis ; drug effects ; Carotenoids ; pharmacology ; therapeutic use ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Gene Expression Regulation, Leukemic ; HL-60 Cells ; Humans ; Leukemia ; drug therapy ; metabolism ; Phytotherapy ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism