Objective: To observe the effect of Sanguisorba tannins on the expression of MGMT (O6-methylguanine-DNA methyltransferase) gene and protein in myelosuppressed mouse model induced by CTX (cyclophosphamide). Methods: Forty Chinese Kunming mice were randomly divided into 4 groups: normal group (NR group), model group (MD group), positive group (G-CSF group) and Sanguisorba tannins group (ST group), with 10 mice in each group. Mouse model of bone marrow suppression was made by intraperitoneal injection of CTX. The mice in G-CSF group were intraperitoneally injected with 30 μg/kg colony-stimulating factor after CTX induction for 3 days. The mice in ST group started to take 20 mg/kg Sanguisorba tannins orally 3 days before CTX induction. The mice in NR group and MD group were given purified water orally. At the end of the treatment, blood and bone marrow of all mice were taken out. Then the flow cytometry technology was used to examine the DNA content in bone marrow cells. The RT-PCR (reverse transcription-PCR) and Western blotting technologies were used to determine the expression changes of MGMT gene and protein in bone marrow cells. Results: Compared with the NR group, the DNA content (P < 0.01) and the expressions of MGMT gene and protein (P < 0.01) were significantly reduced in bone marrow cells of mice in MD group. Compared with the MD group, bone marrow DNA content and the expression levels of MGMT gene and protein were significantly increased in ST group (P < 0.01). Conclusion: Sanguisorba tannins can promote the expression of MGMT gene and protein in bone marrow cells of myelosuppressed mice. Copyright © 2013 by TUMOR.

Objective: To study the mechanism of ziyuglycoside I (ZgI) increasing the number of white blood cells (WBC) in peripheral blood. Methods: KM mice were randomly divided into control group, model group, 3-methyladenosine (3-MA) group, ZgI group (20 mg/kg), and ZgI (20 mg/kg)+3-MA group. On the first day of experiment, myelosuppression mice were induced by ip cyclophosphamide of 120 mg/kg and were continuously gavaged with ZgI for 6 d. On the 6th day, the number of WBC and granulocytes were determined. The degree of hematopoietic stem cells (HSCs) autophagy was observed by transmission electron microscopy and immunofluorescence microscopy. The apoptosis rate of HSCs was determined by flow cytometry, and the expression of Atg5, Atg7, and Beclin-1 proteins were detected by Western blotting. Results: Compared with the model group and 3-methyladenine group, ZgI significantly increased the numbers of leukocyte and granulocytes (P < 0.05), it also significantly stimulated the autophagy of HSCs. Meanwhile, ZgI significantly up-regulated the Atg5, Atg7, and Beclin-1 (P < 0.01) proteins expression in HSCs. Conclusion: The results suggested that ZgI was an efficient autophagic activator, and the mechanism of increasing WBC might be related to the promotion of HSCs autophagy of myelosuppression mice.

Objective: To explore the protective effects of tannins in Sanguisorba Radix (TSR) on myelosuppression mice induced by cyclophosphamide (CTX). Methods: TSR was ig given at the dose of 20 mg/kg for 10 d after ip administration of CTX (200 mg/kg). Results: TSR could significantly increase the numbers of white blood cells, red blood cells, and platelets of myelosuppression in mice. And it could accelerate bone marrow haemopoietic stem/progenitor cells (HSPCs) in myelosuppression mice and enhance cell proliferation by promoting cell cycles from G0/G1 phase to access into S and G2/M phases, then the reduced number of HSPCs induced by CTX was reversed. Moreover, TSR could increase the mRNA and protein expression levels of O(6)-methylguanine-DNA methyltransferase (MGMT) in HSPCs of myelosuppression mice. Conclsion: TSR has a protective function against CTX-induced myelosuppression. The mechanism might be related to protecting hematopoietic stem cells of bone marrow, stimulating hematopoiesis recovery, as well as preventing the apoptosis of hematopoietic stem cells induced by CTX. © 2014 Tianjin Press of Chinese Herbal Medicines.

OBJECTIVETo study the inhibition of berberine (BBR) against ECV-304 apoptosis induced by Staphylococcus aureus (S. aureus).

METHODSECV-304 cells were pre-treated with 128 microg/mL BBR for 2 h and then S. aureus was added (1:100). The viability of cells was detected by MTT (3-4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay. The morphological changes were observed by Hoechst 33258 staining. The protection of BBR for infected cells was detected by DNA Ladder.

RESULTSECV-304 cells' viability were not obviously affected by berberine. But S. aureus induced ECV-304 cells' viability could be significantly inhibited by pre-treatment of BBR (P < 0.05). Besides S. aureus-induced ECV-304 apoptosis could be reduced, with significantly lessened apoptotic body and unobvious DNA degradation.

CONCLUSIONBBR could significantly inhibit S. aureus induced ECV-304 apoptosis.

Apoptosis ; drug effects ; Berberine ; pharmacology ; Cell Line ; Human Umbilical Vein Endothelial Cells ; drug effects ; microbiology ; pathology ; Humans ; Staphylococcus aureus

Objective To explore the effect of blocking BTLA-HVEM (herpesvirus entry mediator-B and T lymphocyte attenuator) pathway on dendritic cell function and the related immunological mechanisms. Methods Murine BTLA extracellular domain eukaryotic expression vector psBTLA was constructed by gene recombination and transfected CHO by Lipofection method. Mouse bone marrow cells were induced to differentiate into DCs by GM-CSF plus IL-4. Expression of BTLA and HVEM on DCs was detected after HSPT0-TC-1 peptide complex stimulation by FACS. Expression of BT-1 and secretion of IL-12 were detected after HSP70-TC-1 peptide complex plus psBTLA transfected CHO culture supernatant stimulation on DCs. Pretreated DCs co-cultured with the same genetic background mouse splenocytes and lymphocytes proliferation and cytokine secretion were detected. Effect of psBTLA gene transfer in vivo on BT-1 expression of DCs and tumor growth on tumor-bearing mice was detected. Results Extracellular domain of murine BTLA was successfully constructed, psBTLA stable transfection CHO cells were obtained and expression of BTLA extracellular domain(sBTLA) was detected the in its culture supernatant. BTLA and HVEM expression of DCs were increased after stimulation by the antigen peptide complex. When DCs were treated with antigen peptide complex plus culture supernatant containing sBTLA, B7-1 expression and IL-12 secretion were increased. Co-cultured with splenocytes, lymphocytes proliferation and cytokine secretion, such as IL-2 and IFN-γ,, were also increased. Gene transfection with psBTLA in vivo promoted B7-1 expression on DCs and inhibited cervical cancer cells growth. Conclusion Blockade of BTLA-HVEM inhibitory pathway with sBTLA can further improve DCs function, activation of lymphocytes and promote antitumor immune response.

OBJECTIVETo investigate the possibility to fabricate a blood vessel scaffold with a combined polymer for tissue engineering.

METHODSA blood vessel scaffold was designed with a combined polymer composed of rabbit vascular smooth muscle cells(VSMCs), collagen and a non-spinning fabric mesh of polyglycolic acid (PGA). VSMCs were implanted into collagen gel and their growth was observed. The mixed solution of VSMCs and collagen was dropped into the tubular scaffold, followed by 7-day culturing.

RESULTSVSMCs formed many prominences after culturing in gelatinous collagen for 3-4 hours. With cells extending, some cells became shuttle- or spindle-shaped. After VSMCs-collagen complex was implanted into the PGA mesh, most of VSMCs remained in the pore of PGA mesh with the formation of gelation. VSMCs could adhere to and grow on the PGA fiber.

CONCLUSIONThe non-spinning PGA porous biodegradable material coated with collagen is a good carrier for VSMCs to adhere and grow.

Animals ; Blood Vessels ; growth & development ; Collagen ; Muscle, Smooth, Vascular ; growth & development ; Polyglycolic Acid ; Polymers ; Rabbits ; Time Factors ; Tissue Engineering ; instrumentation ; methods ; Tissue Scaffolds

Objective To explore the clinical efficacy of focused ultrasound for patients with white lesions of the vulva,as well as its safety and feasibility.Methods Clinical data of 941 patients with white lesions of the vulva treated with focused ultrasound from June 2003 to December 2005 were retrospectively reviewed.The mean age of the patients was 40.8 years(18-70 years)and the median course of the disease was 6.2 years(3 months-45 years).Meanwhile,pathological diagnosis was performed in all the patients before treatment,in which 498 cases were squamous hyperplasia,342 cases were lichen sclerosus and 101 cases were lichen sclerosus with squamous hyperplasia.Patients were followed up and therapeutic effects of focused ultrasound was evaluated at 6 and 12 months after the treatment,respectively.The symptoms of pruritus in the vulva and the changes in the color and elasticity of the vulvar lesions were observed.Results Of all the patients,900 were followed up after the treatment,and the ratio of effectiveness was 94.9%.Only 46 patients(5.1%)had no response to the therapy.Of the effective patients,434 cases were completely cured(48.2%),and 420 cases were improved(46.7%).Pruritus of vulva recurred in 101 patients (11.2%)one year after treatment;however,these patients still had a response to the second or third treatment.Conclusions Focused ultrasound therapy is a highly effective instrument in treatment of white lesions of the vulva.It can not only relieve the symptoms of itching,but is also helpful in recovering the color and elasticity of the vulva.

OBJECTIVETo explore the clinicopathological, immunohistochemical and molecular genetic features of intra-abdomen extra-gastrointestinal stromal tumors (EGISTs) and their differential diagnosis.

METHODSNine cases of EGISTs from the abdominal cavity or retroperitoneum which were previously diagnosed as leiomyoma, leiomyoblastoma, or leiomyosarcoma etc. by a panel of antibodies such as CD117, CD34, alpha-SMA, MSA, desmin, S-100, and PGP9.5 from which five cases were detected for c-kit gene mutation.

RESULTSThe tumors occurred in 5 men and 4 women, the age ranged from 38 to 72 years (mean 61.7 years). Four cases arose from the mesentery, two from omentum, two from retroperitoneum and one located at the hilus of the spleen. The size of tumors ranged from 5 cm to 23 cm (mean 12.9 cm) in diameter and the tumor cell components varied: mainly spindle cells (seven cases), epithelioid cells (one case), mixed cells (one case). Tumors expressed CD117 (8/9), CD34 (5/9), alpha-SMA (3/9), MSA (4/9), desmin (0), S-100 protein (1/9) and PGP9.5 (1/9). Of the five cases examined for heterozygous deletion mutation of 11 exon of the c-kit gene two were found positive. Two borderline cases showed long-term survival of 8 years and 11 years, respectively. In seven malignant cases, two showed adverse outcome, one survived 4 years without recurrence, two were lost in follow up and two new cases were still being in followed.

CONCLUSIONSGIST-type stromal tumors can also occur in the abdomen, most cases were borderline or malignant, tumor coagulative necrosis, mitoses >or= 5 per 50 high-power fields and obvious nuclear atypia indicating malignancy. Differential diagnosis of EGIST including benign or malignant smooth muscle tumors, benign or malignant nerve sheath tumors etc.

Adult ; Aged ; Diagnosis, Differential ; Female ; Humans ; Immunohistochemistry ; Leiomyoma ; pathology ; Leiomyosarcoma ; pathology ; Male ; Middle Aged ; Peritoneal Neoplasms ; chemistry ; genetics ; pathology ; Proto-Oncogene Proteins c-kit ; genetics ; Retroperitoneal Neoplasms ; chemistry ; genetics ; pathology

OBJECTIVETo construct and screen the suppression subtractive hybridization (SSH) library of human renal cell carcinoma (RCC).

METHODSPoly A(+) RNA was isolated from RCC lines 786-O (tester) and renal cell (RC) lines HK-2 (driver), respectively. SSH procedure was performed according to the protocol of the PCR-Select cDNA Subtraction Kit (Clontech), and PCR products were cloned into pT-Adv vector and transformed E. coli TOP10F'. All positive clones picked out were digested and some of which were sequenced.

RESULTSThe SSH library contained 362 clones with SSH cDNA fragments distributed mainly from 0.3 to 0.9 kb. Among 50 clones sequenced randomly, 2 represented unknown genes and the other 48 derived from 36 known genes.

CONCLUSIONThe quality of the SSH library of human RCC is reliable and its construction is the basis for further screening differentially expressed genes of RCC.

Adenocarcinoma, Clear Cell ; genetics ; Cell Line, Tumor ; Gene Library ; Humans ; Kidney Neoplasms ; genetics ; Nucleic Acid Hybridization ; methods