BACKGROUND:Cel replacement therapy as an effective strategy for reconstruction of the central nervous system has very broad application prospects.
OBJECTIVE:To investigate the effect of stereotactic transplantation of neural stem cels into the brain on the neuromotor function of craniocerebral trauma rats.
METHODS:Twenty male Sprague-Dawley rats were equivalently randomized into study and control groups. Animal models of craniocerebral trauma were made using the improved free-fal method in the rats. Then, model rats in the study and control groups were given parenchymal transplantation of embryonic neural stem cels and the same volume of culture medium with no stem cels at 1 day after injury, respectively. Neuromotor function of rats was assessed based on the neurological severity scores. At 2 weeks after transplantation, brain tissues were taken for hematoxylin-eosin staining, anti-BrdU, glial fibrilary acidic protein, β-tubulin III and tyrosine hydroxylase immunohistochemistry staining.
RESULTS AND CONCLUSION:The neurological severity scores in the study group were significantly lower than those in the control group at 1 and 2 weeks after injury (P< 0.05). In the study group, there were many BrdU-positive neural stem cels in the brain tissues, some of which were positive for glial fibrilary acidic protein, β-tubulin III and tyrosine hydroxylase; while in the control group, there was no BrdU-positive cel in the brain tissues. Experimental findings show that neural stem cels stereotacticaly transplanted into the brain can proliferate and differentiate in the brain lesion, and thereby notably improve the neuromotor function of rats with craniocerebral trauma.

OBJECTIVE To screen small molecule compounds from traditional Chinese medicine that can enhance recombinant adeno-associated virus (rAAV) transduction. METHODS Recombinant adeno-associated virus (rAAV) has been established as a powerful tool for in vivo gene transfer and achieved much promise in gene therapy applications. However, widespread clinical use has been limited by transduction efficiency.In the current study,we screened a panel of small molecule compound from traditional Chinese medicine focused on AAV intracellular trafficking process and found salvianolic acid B can significantly enhance rAAV2 transduction. RESULTS Salvianolic acid B caused a dose-depen-dent increase in rAAV2 transduction regardless of vector dose,genome architecture,and over a broad range of cell line from various cell type and species(HEK293,HeLa,HepG2,Huh-7,CHO-K1,LO-2). Salvianolic acid B treatment redirected rAAV2 particles toward large vesicles positive for late endosomal (Rab7)and lysosomal(LAMP1)markers.Furthermore,salvianolic acid B acted to increase accumulation of viral particles at the perinuclear region. CONCLUSION In summary, our results suggest that salvi-anolic acid B redirects rAAV2 toward more productive trafficking pathways and stabilizes perinuclear accumulations of vectors,facilitating productive nuclear trafficking.

OBJECTIVETo investigate the effects of the quinoline derivative PQ1 combined with cisplatin on the proliferation and gap junction communication of prostate cancer PC3 cells.

METHODSWe cultured in vitro prostate cancer PC3 cells and divided them into DMSO blank control, cisplatin control, and cisplatin (10 mg/ml) plus PQ1 (1, 2, 5, 10, and 15 μmol/L) groups. We measured the proliferation of the prostate cancer PC3 cells, determined the expressions of the connexin 43 (Cx43) mRNA and protein by RT-PCR and Western blot, and compared the indexes among different groups.

RESULTSCisplatin combined with PQl at 1 - 10 μmol/L significantly inhibited the proliferation of the PC3 cells and the inhibition rate rose in a concentration- and time-dependent manner, from (48.72 ± 0.98)% vs (50.33 ± 0.62)% at 0 μmol/L to (77.38 ± 1.12)% vs (83.50 ± 1.05)% at 15 μmol/L at 24 and 48 hours (P < 0.05). Compared with the cisplatin control, cisplatin combined with PQ1 at 1, 2, 5, 10, and 15 μmol/L increased the expression of Cx43 mRNA from 0.379 ± 0.113 to 0.669 ± 0.031, 0.831 ± 0. 127, 0.769 ± 0.100, 0.532 ± 0.086, and 0.475 ± 0.134, respectively (P < 0.05), and cisplatin combined with PQ1 at 1, 2, 5, and 10 μmol/L elevated that of Cx43 protein from 0.138 ± 0.146 to 0.263 ± 0.111, 0.306 ± 0.152, 0.415 ± 0.280, and 0.643 ± 0.310, respectively (P < 0.05).

CONCLUSIONThe quinoline derivative PQ1 can promote the gap junction communication of prostate cancer PC3 cells and enhance the killing effect of cisplatin on PC3 cells by upregulating the expressions of Cx43 mRNA and protein.

Aminoquinolines ; pharmacology ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Connexin 43 ; genetics ; metabolism ; Dose-Response Relationship, Drug ; Gap Junctions ; drug effects ; physiology ; Humans ; Male ; Prostatic Neoplasms ; metabolism ; pathology ; physiopathology ; RNA, Messenger ; metabolism ; Time Factors

Background and purpose:It was reported that chemokine receptor CXCR4 and its ligand stromal cell-derived factor 1(SDF-1)were involved in the proliferation,differentiation,and metastasis of tumor.This study was designed to observe the expression of chemokine receptor CXCR4 in hypopharyngeal squamous cell carcinoma tissue and study the relationship between the expression of chemokine receptor CXCR4 and different clinicopathlogical characteristics,and further to explore the clinical significance.Methods:For the detection of the expression of chemokine receptor CXCR4,43 primary hypopharyngeal squamous cell carcinoma tissues,27 normal hypopharyngeal tissues,34 lymph node metastastatic lesions and 9 normal lymph node lesions were detected by immunohistochemical method using rabbit anti-human CXCR4 polyclonal antibody.Results:The positive expression rates of CXCR4 in 43 hypopharyngeal carcinoma tissues and normal tissues were 95.3% and 22.2%,respectively(P

SUMMARY Objective:To investigate the influence of cryoprotectants and cooling rates in vitrificationmethod on the spindles of rabbit M Ⅱ oocytes.Methods:Rabbit oocytes were verified by using cryoloopwith ethylene glycol(EG)singly or EG combined with dimethyl sulphoxide(DMSO)as cryoprotectants,and cooled by taking oocytes directly into liquid nitrogen or by vitrification machine.After frozen rabbit o-ocytes thawed,the microtubulin and chromosome of the spindles were fixation and stained by immunofluo-rescent method.Confocal microscope was used to reveal spindle configuration.Results:In the two proto-cols of single EG used and EG combined with DMSO,the spindles were severely injured.But in protocolof EG combined with DMSO and at ultra-rapid cooling rate,the normal configuration of spindle rate ofthawed rabbit oobytes was similar to that of the control group.Conclusion:The protocol of EG combinedwith DMSO as cryoprotectants and with extremely high cooling rate by vitrification machine can producethe best effect on conservation of spindle configuration in vitrification of rabbit oocytes.

Background and purpose:Lung cancer is thought to be caused by multiple-step carcinogenesis. Identification of the genetic alterations that occur in tumors is an important approach to understanding carcinogenesis. We identified chromosomal abnormality in lung cancer by the molecular cytogenetic techniques of comparative genomic hybridisation(CGH),the technology could help to comprehend the relationship between chromosome abnormality, different patho-types,and clinical features of lung cancer.Methods:CGH was used to detect the global genomic aberration in the fresh cancer tissue cells from 30 patients with lung cancer.Results:Chromosomal abnormality were detected in all of 30 cases with lung cancer,the altofrequent gains in 1p11-p22,5p11-p14,16p 11-P12,19q13, 19p 13,20p12,21q21 and the altofrequent losses in 5q,6p24-pter,9p31-qter,13q21-qter,14q21-qter were found in all three types of lung cancer,the marked differences of chromosomal abnormalities in three types of lung cancer were also found.Conclusions:The cytogenetic aberration exists generally in lung cancer cells,the cytogenetic aberration is the base of the initiation and progression of the lung cancer.There are some different chromosomal abnormalities between different types of lung cancer,which may serve as a marker to differential diagnosos of the three types of lung cancer.As to the progression of malignant neoplastic disease,the complexity of chromosomal abnormality is obviously elevated.Different carcinogenic agents(smoking for example)may induce different chromosomal abnormalities.

Objective: To assess the effects of subchronic intake of high-dose Dendrobium officinale on body weight, food intake, food utilization, and blood biochemical parameters in rats, so as to provide insights into assessment of edible safety of D. officinale. Methods: Eighty SPF-grade SD rats were randomly divided into the low-, medium- and high-dose groups and the control group, of 10 male and 10 female rats in each group. Rats in the low-, medium- and high-dose groups were administered with D. officinale feeds at doses of 2.0, 4.0, and 8.0 g/kg body weight, respectively, while animals in the control group were given basic diet for successive 13 weeks. The rat body weight, food intake, food utilization, and blood biochemical parameters were compared between groups. Results: Normal diet and activity was seen in all rats, and no abnormal syndromes, signs or deaths were found during the study. There were no significant differences in rat body weight, food intake, total weight gain, total food intake, alanine aminotransferase, aspartate aminotransferase, urea nitrogen, creatinine, triacylglycerol, cholesterol, total protein, albumin, albumin/globulin ratio or blood glucose among four groups (P>0.05). The food utilization 7 weeks post-administration [(8.71%±0.78%) vs. (10.54%±1.37%), P<0.05] and the total food utilization [(18.00%±0.41%) vs. (19.51%±1.21%), P<0.05] were significantly lower in male rats in the high-dose group than in the control group. Conclusion Subchronic intake of high-dose D. officinale shows no toxicity in rats, and reduced food utilization may be associated with the health function of D. officinale in male rats.

Objective To evaluate the roles of magnetic resonance imaging and proton magnetic resonance spectroscopy(1H-MRS) in the diagnosis of meningiomas. Methods 98 patients with meningiomas underwent conventional pre-contrast MR and contrast MR. Among them, 28 cases had two dimensional single voxel or multi voxel 1 H-MRS simultaneously both in the lesion's region and the contralateral side. Results On precontrast MR images of 98 cases, T1 WI showed 58.1% (61/105) isointensities, 31.4% (33/105) faintly low intensities and 10. 5% (11/105) mixed intensities; T2WI showed 40. 0% (42/105) isointensities, 41.0%(43/105) hyperintensities, 10.5% (11/105) faintly low intensities and 8.5% (9/105) mixed intensities. After administration of Gd-DTPA, the solid part of the tumors exhibited various enhancement in all the 98 cases.28 cases of MRS exhibited specific different spectral peaks, including increased of choline-containing compounds(Cho), absent or decreased of acetylaspartate(NAA), and the unchanged of creatine(Cr). The value of NAA, Cr, Cho, NAA/Cr, Cho/Cr, NAA/Cho in the tumor center of meningioma were 0. 09 ± 0.06,0.31 ± 0. 22, 0.46 ± 0. 16, 0.33 ± 0. 42, 1.50 ± 0. 68, 0. 15 ± 0.08, compared with the contralateral normal region, Cr has no significant difference (P ＞ 0. 05), NAA, Cho, NAA/Cr, Cho/Cr, NAA/Cho had significantly differences(P ＜ 0.05). Conclusion Conventional pre-contrast MR and contrast MR is the most important dignostic means for meningiomas, 1H-MRS combined with MRI can improve the diagnostic accuracy of meningiomas.

Cysticercosis is caused by the metacestode form of Taenia solium-Cysticercus cellulosae and it causes great economic losses and threatens the people's health. There are some problems on how to control cysticercosis, in order to resolve the key problem that the native antigen to diagnose and prevent cysticercosis is very limited and is not satisfied, Pichia pastoris Expression System was used to express recombinant P2 protein. The interested P2 gene was got by digesting the pGEM - P2 vector using restriction endonuclease, then it was inserted into the secretory pPIC9K Pichia pastoris expression vector and transformed into E. coli. Positive recombinant plasmids were selected sequenced and named pPIC9K-P2 and it was linearized by Sal I and Bgl II, then the linear DNA transfored into Pichia pastoris GS115 by electroporation. The recombinant expression vector pPIC9K - P2 integrated into GS115 via homologous recombination between the transforming DNA and regions of homology within the genome. The transformants were screened for multicopy recombinants using G418 and were distinguished for Mut phenotypes by MD and MM. Two different phenotypes were generated-HIS+ MUT+ (Methanol utilization plus) and HIS+ MUT(S) (Methanol utilization slow). PCR analysis of the multicopy recombinants indicated that the P2 gene was integrated within the genome of pichia Pastoris. The multicopy recombinants were named GS115/pPIC9K - P2HIS+ MUT+ and GS115/pPIC9K-P2HIS+ MUT(S), both HIS+ MUT+ and HIS+ MUT(S) were induced with methanol. The results of SDS-PAGE and Western blot demonstrated that the culture supernatant of the induced Pichia pastoris contained P2 protein which was accumulated up to 33 % of total proteins in the culture supernant and its molecular weight is 12.6kD. The results of the clinical study indicated that the expression P2 protein could be used to diagnose human cysticercosis and swine cysticercosis as diagnosis antigen.
Animals ; Blotting, Western ; Cysticercosis ; diagnosis ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Genome, Fungal ; genetics ; Helminth Proteins ; genetics ; metabolism ; Humans ; Phosphoproteins ; genetics ; metabolism ; Pichia ; Polymerase Chain Reaction ; Recombinant Proteins ; genetics ; metabolism ; Swine ; Taenia ; metabolism

Background and purpose:Thyroid carcinoma is a common endocrine tumor with an incidence that has increased over recent decades. The aim of the present study was to investigate the effectiveness of carbon nano-par-ticles-labeled lymph nodes in neck dissection for papillary thyroid cancer (PTC), focusing on the protectiveness for the recurrent laryngeal nerve (RLN) and parathyroid glands.Methods:Forty-eight patients with PTC treated from Apr. to Aug. 2015 were randomly divided into two groups. Group A patients (24 patients) were treated with lobectomy/total thyroidectomy plus unilateral/bilateral central lymph node dissection by conventional meticulous capsular dissection technique; Group B patients (24 patients) were treated with the same surgical procedures as group A, 5 min after the injection of carbon nano-particles. The operative time, intra-operative blood loss, incidence of RLN injury, incidence of transient hypocalcemia, the number of total lymph nodes and the ratio of metastatic nodes were collected and analyzed. Results:For unilateral lobectomy, the number of lymph nodes in group B was signiifcantly greater than that in group A (P<0.05). For total thyroidectomy, the operative time, and the incidence of transient hypocalcemia in group B were both lower than those in group A (P<0.05), and the number of total lymph nodes was signiifcantly higher than that in group A (P<0.05). In group B, the ratio of metastatic nodes were 26.7% (unilateral) and 33.3% (bilateral) in stained lymphnodes, and 11.8% and 25.9% in non-stained lymph nodes.Conclusion:The carbon nano-particles-labeled lymph nodes in neck dissection could facilitate to protect parathyroids and increase the number of lymph nodes, especially in total thyroidectomy plus bilateral central lymph node dissection.