OBJECTIVETo investigate the apoptosis and proliferation effect of matrine on human medulloblastoma cell line D341 in vitro and the effect of the expression of the related caspase 3 and caspase 9 proteins.

METHODSThe D341 cells were cultivated successfully in vitro. Then the cells were divided into 5 groups according to the concentration of matrine (0.5 mg/mI group, 1.0 mg/ml group, 1.5 mg/ml group, 2.0 mg/ml group and the control group was 0 mg/ml). All the experiments were repeated three times. The cell morphologic and structure change was observed with the optical microscope and the transmission electron microscope. The proliferation of D341 cell was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining. The expression of Caspase3 and Caspase9 was detected by Western blot.

RESULTSWith the effect of matrine, the proliferation inhibition rate gradually increased with drug concentrations increasing, and there was a significant difference (P < 0.01). The inhibitory effect of matrine on cell proliferation was different with the different treatment time, there was a significant difference between the 24 h to 72 h groups (P < 0.01). The apoptotic rate increased with matrine concentrations increasing. There were significant differences between the group of 0.5 mg/mI or 1.0 mg/mI to the group of 1.5 mg/mI or 2.0 mg/mI (P < 0.05). The apoptotic rate increased with the prolonged treatment time. There were significant differences between the group of 24 h or 48 h to the group of 72 h ( P < 0.05). With the increase of matrine concentration, the expression of Caspase 3 and Caspase 9 increased (P < 0.01).

CONCLUSIONMatrine induces the apoptosis, and inhibits the proliferation of human medulloblastoma D341 cells in vitro by up-regulation of the expression level of Caspase3, Caspase9.

Alkaloids ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; metabolism ; pathology ; Humans ; Medulloblastoma ; metabolism ; pathology ; Quinolizines ; pharmacology ; Up-Regulation

Objective To explore the change of plasma orexin A concentration and the correlation between plasma orexin A concentration and energy intake in obese children.Methods Fasting plasma orexin A concentrations,boaly mass index(BMI) and energy intake were measured in 48 obese children(obese group) and 48 healthy children(healthy control group),and these indexes were compared,the correlation between plasma orexin A concentration and BMI,energy intake were analyzed.Results 1.The plasma orexin A concentration in obese group was significantly lower than that in healthy control group(F=5.632 P=0.008).2.In obsess group,there were negative correlation between plasma orexin A concentration and BMI(r=-0.478 P=0.012),positive correlation between plasma orexin A concentration and total energy intake(r=0.503 P=0.007),fat intake(r=0.659 P=0.006) and protein intake(r=0.381 P=0.026),and there was negative correlation between plasma orexin A concentration and carbohydrate(r=-0.316 P=0.022).3.In healthy control group,there were negative correlation between plasma orexin A concentration and BMI(r=-0.491 P=0.018),positive correlation between plasma orexin A concentration and total energy intake(r=0.512 P=0.009),fat intake(r=0.406 P=0.005),protein intake(r=0.313 P=0.020),and carbohydrate(r=0.432 P=0.025).Conclusions Orexin A may be involved in regulation of energy metabolism in obese children,and the interaction between plasma orexin A and energy intake might be different in different nutritional status in children.

A method of LC-QTOF/MS combining with chemical synthesis has been used to determine the structures of three novel bile acids from bear bile powder. Reference substances of tauroursodeoxycholic acid and taurochenodeoxycholic acid were oxidized by pyridinium chlorochromate. The products were analyzed by LC-QTOF/MS. Total 4 products including 3 isomers were predicted and identified according to the PCC oxidation theory and LC-QTOF/MS results. Bear bile powder samples were dissolved by methanol and analyzed by LC-QTOF/MS. Three unknown peaks were found and identified as 2-[[(3beta, 5beta)-3-hydroxy-7, 24-dioxocholan-24-yl]amino]-ethanesulfonic acid, 2-[[(5beta)-3, 7, 24-trioxocholan-24-yl]amino]-ethanesulfonic acid and 2-[[(5beta, 7beta)-7-hydroxy-3, 24-dioxocholan-24-yl]amino]-ethanesulfonic acid, separately, by matching their results with that of oxidation products above.
Animals ; Bile ; chemistry ; Bile Acids and Salts ; analysis ; chemistry ; Chromatography, Liquid ; methods ; Isomerism ; Molecular Structure ; Oxidation-Reduction ; Powders ; chemistry ; Spectrometry, Mass, Electrospray Ionization ; methods ; Taurochenodeoxycholic Acid ; chemistry ; Ursidae

Based on the literature study,the thoughts and possible therapeutic mechanism in treating male infertility from the perspective of spleen and kidney by regulating intestinal flora were explored.Disturbance of intestinal flora is one of the important factors leading to the development of male infertility,and the spleen and kidney have certain similarities to intestinal flora in the physiological function and pathological changes.Moreover,tonifying the kidney and strengthening the spleen can regulate the intestinal flora by fostering the growth of beneficial bacteria,inhibiting the reproduction of pathogenic bacteria,and protecting the barrier of the intestinal mucosa.Therefore,the possible therapeutic mechanisms in treating male infertility with the prescriptions for tonifying the kidney and strengthening the spleen to regulate intestinal flora are as follows:inhibiting the expression of inflammatory factors to reduce the inflammatory reaction of testicular tissues;improving the antioxidant capacity to alleviate the damage of spermatozoa caused by oxidative stress,and improving the bad mood to alleviate the impact of psychological stress on the reproductive system.The exploration of the thoughts for treating male infertility from the perspective of spleen and kidney by regulating intestinal flora may provide a new entry point for modern Chinese medicine clinical treatment of male infertility.

The study aimed to explore the changes of the immunocyte quantities and cytotoxicity in bone marrow, and how many hematopoietic progenitor cells retained after the bone marrow cells were activated by different combinations of various cytokines. Bone marrow cells were divided into four groups and were cultured in vitro: (1) Control: no cytokines were added. (2) IL-2 group: bone marrow cells were activated by IL-2. (3) CD3-AK group: bone marrow cells were activated by IL-2 and CD3-McAb. (4) CIK group: IFN-gamma, IL-1, IL-2 and CD3-McAb were added. CFU-GM assay and CD3 phenotype detection were performed before and after activating culture in all groups. The changes of cell quantities during culture and cytotoxicity of cultured cells were tested. CD3 positive cells markedly increased in both CD3-AK and CIK groups. The cell numbers and cytotoxicity of CD3-AK and CIK groups were higher than those of control or IL-2 group obviously after culture (P < 0.05). CFU-GM were decreased in all groups after culture and there had no significant difference among four groups. The combination of IL-2 and CD3-McAb not only stimulates the proliferation of marrow immunocytes and increases their cytotoxicity but retains enough hematopoietic progenitor cells as well. This combination of cytokines can be used to purge autologous bone marrow in vitro.

A rapid method of Liquid chromatography-quadrupole time-of-flight mass spectrometry (LC-Q-TOF-MS) combined with pyridinium chlorochromate (PCC) oxidation has been developed to determine chemical structures of two novel isomers in bear bile powder. Derivatives of ursodeoxycholic acid (UDCA) and chenodeoxycholic acid (CDCA) were semi-synthesized by PCC oxidation, then were analyzed by LC-Q-TOF-MS. Separation was carried out on a reverse column with the mobile phase of acetonitrile-0.1% formic acid (45:55). The data of Q-TOF-MS was acquired by MS, MS/MS, positive and negative modes. Since UDCA and CDCA were stereochemical isomeric at an alcohol position, two oxidation products were same and have been confirmed by LC-Q-TOF-MS. Other two products were also determined based on the PCC oxidation theory. Samples of bear bile powder were dissolved by methanol and measured by LC-Q-TOF-MS. Two unknown peaks were found and identified by matching their retention times and accurate mass spectra ions with PCC oxidation productS. Finally, the structures of two new bile acids in bear bile powder were confirmed as 3alpha-hydroxy-7-oxo-5beta-cholanic acid, 7alpha-hydroxy-3-oxo-5beta-cholanic acid, respectively.
Animals ; Bile ; chemistry ; Chromatography, Liquid ; methods ; Isomerism ; Oxidation-Reduction ; Powders ; chemistry ; Pyridinium Compounds ; chemistry ; Tandem Mass Spectrometry ; methods ; Tissue Extracts ; chemistry ; Ursidae ; Ursodeoxycholic Acid ; chemistry

To study immunophenotype and cytotoxicity of the immunocytes in bone marrow and peripheral blood after activation by combined cytokines, mononuclear cells (MNC) of bone marrow and peripheral blood were activated by IFN-gamma, IL-1, IL-2 and McAb-CD3 in vitro. The cell amount and morphology during culture were observed. Cytochemical staining and immunophenotype analysis were done before and after culture in two groups of MNC. Cytotoxicity was tested by MTT method. The results showed that the cell number of two groups increased obviously in culture (P < 0.05), while the peripheral blood mononuclear cells increased more markedly (P < 0.05). The cytochemical staining showed POX decrease, but PAS increase in two groups. The positive ratios of CD3(+), CD56(+) and CD38(+) cells in two groups increased obviously after culture (P < 0.05), but there was no significant difference between those two groups. CD3(+) CD56(+) cells increased obviously in peripheral blood mononuclear cells activated by cytokines (P < 0.05), but CD3(+) CD56(+) cells did not increase in bone marrow mononuclear cells. There was no significant difference between two groups' cytotoxicity. It was concluded that IFN-gamma, IL-1, IL-2 and McAb-C D3 increased cell number and cytotoxicity of both bone marrow and peripheral blood mononuclear cells that can be used in cell immunotherapy.
ADP-ribosyl Cyclase ; immunology ; ADP-ribosyl Cyclase 1 ; Antibodies, Monoclonal ; pharmacology ; Antigens, CD ; immunology ; Bone Marrow Cells ; cytology ; drug effects ; immunology ; CD3 Complex ; immunology ; CD56 Antigen ; immunology ; Cell Count ; Cell Division ; drug effects ; Coculture Techniques ; Cytokines ; pharmacology ; Cytotoxicity Tests, Immunologic ; Cytotoxicity, Immunologic ; drug effects ; HL-60 Cells ; Humans ; Immunophenotyping ; Interferon-gamma ; pharmacology ; Interleukin-1 ; pharmacology ; Interleukin-2 ; pharmacology ; K562 Cells ; Leukocytes, Mononuclear ; cytology ; drug effects ; immunology ; Membrane Glycoproteins ; Time Factors

It has need to separate red blood cells (RBC) from marrow graft in ABO group unmatched BMT and auto-BMT with purging tumor cells, the separating effect of methylcellulose was observed. The mixture of 0.5% methylcellulose and bone marrow was laid up in an open transfusion system, and then sedimentation of RBC was performed in the transfusion tube. The separating results of 18 marrow grafts showed that the recovery rates of mononuclear cells and CD34(+) cells were (83.8 +/- 55.2)% and (90.3 +/- 7.2)%, respectively. RBC residual rate was (4.3 +/- 1.5)%. The yield of CFU-GM was (60.8 +/- 22.4)/2 x 10(5) MNC, and there was no difference to [(69.8 +/- 23.4)/2 x 10(5) MNC] yielded from same marrow samples, separated by Ficoll-Hypaque separation. It is concluded that this method could be used for bone marrow transplantation.
Bone Marrow Transplantation ; methods ; Cell Separation ; methods ; Erythrocytes ; immunology ; Humans ; Methylcellulose ; pharmacology

OBJECTIVETo investigate the clinical features, treatment and prognosis of the young adult papillary thyroid cancer with bilateral cervical lymph nodes metastases.

METHODSTwenty-four papillary thyroid cancer patients (mean age 16.6 year-old, range 11 to 20 year-old) with bilateral cervical metastases from January 1, 1970 to December 31, 1985 were analyzed.

RESULTSThe proportion of the patients in the contemporary patients of the corresponding period was 34.8% (24/69). All the patients in the group were followed up at least 20 years. The survival rates of the patients at 10 years and 20 years were 100% and 90.9%. The recurrence rates of local tumor and distant tumor (lung metastases) were 20.8% (5/24) and 12.5% (3/24).

CONCLUSIONAfter a radical surgery, the recurrence rate of young adult thyroid papillary cancer patients with bilateral cervical metastases is relatively high and the patients have well prognosis.

Adolescent ; Adult ; Carcinoma, Papillary ; pathology ; surgery ; Child ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Neoplasm Recurrence, Local ; Prognosis ; Retrospective Studies ; Thyroid Neoplasms ; pathology ; surgery ; Thyroidectomy

Objective To investigate the magnetic resonance multimodal function imaging characteristics of small hepa-tocellular carcinoma and evaluate its diagnostic value in small liver cancer. Methods Thirty-nine patients with small hepato-cellular carcinoma were selected in the First Affiliated Hospital of Xinxiang Medical University from January 2016 to January 2018. The magnetic resonance plain scan data,dynamic enhance data and diffusion imaging data of all patients were analyzed. Results Forty-five lesions were found by plain scan,and 48 lesions(37 lesions were rapid rise and rapid descent type,6 le-sions were flatbed type,3 lesions were slow ascent and rapid descent type,2 lesions were not obvious strengthening type)were found by dynamic enhancement. The lesions were hyperintense or slightly hyperintense on the diffusion weighted imaging (DWI)image. The lesions were hyperintense or slightly hyperintense on the DWI. The main shape of lesions was round,irregu-lar ring and nodositas. The apparent diffusion coefficient(ADC)value of cancer tissue and normal liver tissue gradually de-creased with the increase of b value. Under the same b value,the ADC values of small hepatocellular carcinoma was significant-ly lower than that in the normal liver tissue(P < 0. 05). Conclusion Magnetic resonance multimodality functional imaging (magnetic resonance imaging dynamic enhanced combine with multi-b value DWI)can effectively improve the diagnostic effi-cacy of small hepatocellular carcinoma.