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.

OBJECTIVE: To explore the effect of LRRC4 on the mobility and invasion of glioblastomas U251 cells through the SDF-1alpha/CXCR4 axis. METHODS: RT-PCR, transfilter cell invasion assay, adhesion assay, scraping test, scrape loading, and dye transfer assay were used to determine the effect of LRRC4 on U251 cells. RESULTS: SDF-1 alpha could increase the invasion in U251 which expressed CXCR4. The reintroduction of LRRC4 in U251 cells could inhibit the expression of CXCR4. LRRC4 also inhibited the adhesion ability of U251 to ECV304 as well as the mobility and invasion ability in vitro, which was mediated by the SDF-1alpha/CXCR4 axis. Furthermore, LRRC4 could greatly enhance the gap junctional intercellular communication of U251 cells. CONCLUSION The reintroduction of LRRC4 in U251 cells can inhibit the expression of CXCR4 and the SDF-1alpha/CXCR4 axis-mediated cell invasion in vitro.
Cell Adhesion ; Cell Line, Tumor ; Cell Movement ; Chemokine CXCL12 ; metabolism ; Glioblastoma ; pathology ; Humans ; Neoplasm Invasiveness ; Nerve Tissue Proteins ; genetics ; Receptors, CXCR4 ; metabolism

OBJECTIVETo obtain the monoclonal antibody against hexon protein of human adenovirus.

METHODSBALB/c mice were immunized with purified recombinant hexon protein, and the spleen cells of the mice were isolated and fused with myloma cells. Four hybridoma cell strains were screened by indirect ELISA and cultured, and the sensitivity, specificity and virus neutralizing activity were analyzed with ELISA, Western blotting and neutralizing test.

RESULTSThe mouse ascites produced by these hybridoma cells contained specific monoclonal antibodies against hexon protein of human adenovirus as identified by ELISA and Western blot, and the antibody generated by 4C6 strain showed human adenovirus type 3-neutralizing activity.

CONCLUSIONThe monoclonal antibodies against hexon protein with high specificity have been successfully obtained, and these antibodies can be useful in developing assays for early diagnosis of HAdV3 infection and also in study of therapeutic drugs of the infection.

Adenoviruses, Human ; chemistry ; immunology ; Animals ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibodies, Viral ; biosynthesis ; immunology ; Blotting, Western ; Capsid Proteins ; biosynthesis ; genetics ; immunology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; genetics ; Humans ; Hybridomas ; secretion ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; biosynthesis ; immunology

Objective To select the optimum process of granule spray for Tangwang Mingmu Granules, To improve the yield and finished product quality. Methods The effects of the relative density, the type of excipients, the ratio of the extract and the excipient, the inlet temperature-the material temperature, the spray rate and the atomization pressure were selected by single factor and orthogonal test respectively to determine the optimum preparation process. Validation for three batch sample, particle flow, hygroscopicity and other related content of the preparation were studied. Results The optimum spray granulation process of Tangwang Mingmu Granules: the relative density of extract (60 ℃ determination) was 1.15–1.20; the excipient was selected as dextrin; the ratio of dry extract and excipient was 58:42; inlet temperature - material temperature was 90-75 ℃; atomization pressure was 0.12 mPa;spray rate was 10 Hz.Conclusion The preparation technology of Tangwang Mingmu Granules optimized in this study is with high preparation rate, good fluidity and hygroscopicity, which can be used for the preparation of Tangwang Mingmu Granules.

Objective To explore surgical treatment of gliomas involving the motor eloquent area. Methods Twelve cases of gliomas involving precentral gyrus were underwent awake surgery procedures assis- ted with neuronavigation and brain functional mapping by cortical electrical stimulation.Results Eleven ca- ses acquired accurate location of both lesions and eloquent areas by neuronavigation and direct cortical stimula- tion.7 cases of motor cortices and 2 cases of motor speech centers were confirmed during the operation.Re- section,verified by postoperative MRI,was total in 8 cases (66.7%) and subtotal in 4 patients.Histological examination revealed an infiltrative glioma in all cases (8 low grade astrocytomas,2 high grade astrocytomas and 2 glioblastoma).Four patients had no postoperative deficit,while the other 8 patients were impaired, with,in all cases except one,complete recovery in 7 days to one month.Conclusion Direct cortical elec- trical stimulations and awake surgery offer a reliable,precise and safe method,allowing functional mapping es- pecially useful in case of infiltrative cerebral tumors in eloquent areas.This technique allows improvement in the quality of tumoral resection and concurrently a minimization of the risk of definitive postoperative neurologi- cal deficit.

Objective To establish a simple and specific method for the determination of rhaponticin in Rhei Radix et Rhizoma. Methods A comparison of the methods of polyamide, silica gel TLC and HPLC in the determination of rhaponticin was studied to investigate the optimum way to identify authentic Rhei Radix et Rhizoma. Results The durability of polyamide and silica gel TLC method for the determination of rhaponticin in Rhei Radix et Rhizoma was poor. HPLC method was more suitable for the detection of rhaponticin. Conclusion HPLC method is accurate and reliable, and can be used for the quality control of Rhei Radix et Rhizoma and provide references for the authenticity of Rhei Radix et Rhizoma.

OBJECTIVETo investigate the regularity of changes in the main active components of Schisandra chinensis preparations before and after being processed with vinegar.

METHODHigh performance liquid chromatography was used to analyze the changes in the content of lignans and organic acids in S. chinensis preparations before and after being processed with vinegar.

RESULTAfter being processed with vinegar, the content of lignans in S. chinensis preparations significantly reduced, while that of organic acids significantly increased. The ratio between them reduced from 1:16 (raw product) to 1:21 (vinegar products).

CONCLUSIONThe changes in the main pharmacological effects of S. chinensis are related to not only the changes in the contents of lignan and organic acids, but also the significant changes in the ratio relations between the two components. This is of great significance to the correlation among the principle of S. chinensis processed with vinegar and material basis and the changes in pharmacological effects.

Acetic Acid ; Acids ; analysis ; isolation & purification ; Chemical Fractionation ; Chemistry, Pharmaceutical ; Drugs, Chinese Herbal ; analysis ; isolation & purification ; Lignans ; analysis ; isolation & purification ; Schisandra ; chemistry

Objective To understand mental experience of first diagnosed young tuberculosis patients and provide basis for clinical nursing. Methods To make depth interviews with 12 first diagnosed tuberculosis patients through phenomenological method, to extract the theme by analyzing the data. Results First diagnosed tuberculosis patient got deny, anxiety and fear in their heart and a series of emotional changes, increased demand for access to information, greater economic pressure in the process of chemotherapy, increased demand of social support. Conclusions we should concern mental status of first diagnosed young tuberculosis patient, to provide timely medical care, information, emotion help, economic and social support to alleviate the stress response and improve treatment effect.

Objective To explore affection of family' s intimate situation and suitability on life quality of patients suffered from hepatocirrhosis in later stage. Methods The patients suffered from hepatocirrhosis in later stage were investigated by the family' s intimate situation and suitability measurement scale in Chinese version (FACESII-CV) as the measurement tool for the classification of group based on self-family' s intimate situation and suitability as actual feeling assessment and calculation of marks. Choosing the family' s intimate situation mark assessment ≥65, suitability ≥ 53 to be the observation group (80 cases), the family's intimate situation mark assessment ＜ 65, suitability ＜ 53 to be the control group (76 cases). Psychological and life quality in two groups were assessed by living quality assessment measure form (GQOLI-74) and feeling measure form. Results the life quality and psychological quality in the observation group is evidently higher than those in the control group, and the differences were statistically significant (P ＜ 0. 01). Conclusions Family' s intimate situation for the patients suffered from hepatocirrhosis of later stage are required for more care so as to take social psychological interference to the patients, improve suitability and promote the health recovery to reach the high life quality of patients.

OBJECTIVE: To examine the expression absence of LRRC4 gene in glioblastoma cell lines. METHODS: RT-PCR and Northern blot were used to detect the expression of LRRC4 gene in 6 glioblastomas cells lines. Polymerase chain reaction and DNA sequencing were used to screen the LRRC4 gene mutation, while bioinformation assay was used to search for the reason of LRRC4 gene absence in U251 cell lines. RESULTS: The expression of LRRC4 was absent in 6 malignant glioma cell lines (U251, U87, BT325, SF126, SF767 and M17), which were examined by Northern-blot and RT-PCR assay. All sequencing of PCR products from gDNA of SF126, SF767, and M17 cell lines contained the point mutation at the same position ( LRRC4 geneT977A) (3/5), which was a synonymous mutation. However, PCR products from gDNA of U251 and U87 cell lines (2/5) were not obtained. The expression absence of LRRC4 was ascribed to the loss of homozygosity of 7q32-ter in U251 cell lines. CONCLUSION The expression of LRRC4 gene is absent in glioblastoma cell lines, and it offers the important experiment proof for LRRC4 to act as a new candidate of brain tumor suppressor gene from glioma. The loss of homozygosity of 7q32-ter contributed to the expression absence of LRRC4 in U251 cell lines.
Base Sequence ; Blotting, Northern ; Brain Neoplasms ; genetics ; pathology ; Cell Line, Tumor ; DNA Mutational Analysis ; Gene Expression Regulation, Neoplastic ; Glioblastoma ; genetics ; pathology ; Humans ; Nerve Tissue Proteins ; genetics ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction