BACKGROUND: Vascular endothelial growth factor (VEGF) is able to effectively treat the ischemic heart disease, but in vivo VEGF cannot be maintained effective concentration. OBJECTIVE: To detect the expression of VEGF mRNA and protein in human bone marrow mensenchymal stem cells transferred by VEGF-165 gene. DESIGN, TIME AND SETTING: The empirical study was conducted from March 2006 to April 2007 at Shanghai Chest Hospital. MATERIALS: Human bone marrow mesenchymal stem cell line and VEGF were offered by Basic Laboratory, Thoracic Tumor Institute, Shanghai Chest Hospital. METHODS: hVEGF165 gene was reconstructed in pcPGK-vector and transferred into human bone marrow mensenchymal stem cells (BMSCs) by liposome-mediated method, clone screening by G418. MAIN OUTCOME MEASURES: The mRNA and protein of VEGF gene in transferred cells was detected by reverse transcription-polymerase chain reaction (RT-PCR), Real time PCR, Western Blot, enzyme linked immunosorbent assay (ELISA) in 3 stem cells of pcPGK-VEGF165-IRES-GFP and pcPGK- IRES-GFP, respectively. RESULTS: pcPGK-hVEGF165 vector was reconstructed and transferred into hMSCs successfully. The expression of hVEGF165 in the transfected hMSCs was demonstrated with RT-PCR and Real time PCR. Western Blot and ELISA demonstrated that the expression of hVEGF165 in the transfected hMSCs and VEGF protein in supernatant were significantly more than untransfected hMSCs. CONCLUSION: hVEGF165 can be successfully transfected into BMSCs by using liposome mediated gene transfer. Stably expressed VEGF165 cell line can be obtained..

Objective　To study the telomerase activity in blood cancer cells by TRAP for monitoring tumor metastasis in blood. Methods　Twenty-five patients with lung cancer surgically treated and 35 patients before chemotherapy were determined for telomerase activity of cancer cells in the blood by TRAP, and 30 patients with non-tumor diseases as control. Results　In the operative group,13 patients(52％) showed telomerase activity in pulmonary artery blood during operation, which was much higher than that of peripheral blood before operation (24％，P＜0.05). Telomerase positive rate was significantly higher in stage Ⅲ to Ⅳ (64％) than that in stage Ⅰ to Ⅱ (30％,P＜0.05).In the peripheral blood of patients with non-tumor diseases, telomerase was negative. Conclusion　Telomerase activity may be an indicator for detecting lung cancer cells in peripheral blood as well as tumor metastasis and relapse.

Objective To explore the clinical effect of balanced cupping intervention in treatment of patients with primary dysmenorrhea differentiated as type of qi stagnation and blood stasis.Methods A total of 64 patients with primary dysmenorrhea differentiated as type of qi stagnation and blood stasis in the First Affiliated Hospital of Henan University of Chinese Medicine from January 2020 to December 2021 were selected as the research objects,and they were divided into observation group and control group according to the order of consultation and patients'wishes,with 32 cases in each group.The control group received routine health guidance for dysmenorrhea,while the observation group received balanced cupping intervention on the basis of the control group.The Visual Analogue Scale(VAS)score,dysmenorrhea symptom score,serum prostaglandin E2(PGE2),estradiol(E2),menstrual on-set time,menstrual cycle,menstrual volume,and efficacy were compared between the two groups be-fore and after treatment.Results There were significant differences in VAS scores during the 1st,2nd and 3rd menstruation compared with those before intervention between the two groups(P＜0.05);the VAS scores of the observation group during the 1st,2nd and 3rd menstruation were significantly lower than those of the control group(P＜0.05).There were significant differences in dysmenorrhea symptom scores during the 1st,2nd and 3rd menstruation with those before intervention between the two groups(P＜0.05);the dysmenorrhea symptom score of the observation group during the 1st,2nd and 3rd menstruation was significantly lower than those of the control group(P＜0.05).The total effective rate of the observation group was significantly higher than that of the control group(P＜0.05).There were significant differences in PGE2 and E2 levels during the 1st,2nd and 3rd menstruation with those before intervention between the two groups(P＜0.05).There were significant differ-ences in PGE2 and E2 levels between the two groups during the 1st,2nd and 3rd menstruation(P＜0.05).There were no significant differences in menstrual cycle,menstrual duration and menstrual volume between the two groups(P＞0.05).Conclusion Balanced cupping intervention has a good effect and high safety in the treatment of patients with primary dysmenorrhea differentiated as type of qi stagnation and blood stasis.

Objective To explore the clinical effect of balanced cupping intervention in treatment of patients with primary dysmenorrhea differentiated as type of qi stagnation and blood stasis.Methods A total of 64 patients with primary dysmenorrhea differentiated as type of qi stagnation and blood stasis in the First Affiliated Hospital of Henan University of Chinese Medicine from January 2020 to December 2021 were selected as the research objects,and they were divided into observation group and control group according to the order of consultation and patients'wishes,with 32 cases in each group.The control group received routine health guidance for dysmenorrhea,while the observation group received balanced cupping intervention on the basis of the control group.The Visual Analogue Scale(VAS)score,dysmenorrhea symptom score,serum prostaglandin E2(PGE2),estradiol(E2),menstrual on-set time,menstrual cycle,menstrual volume,and efficacy were compared between the two groups be-fore and after treatment.Results There were significant differences in VAS scores during the 1st,2nd and 3rd menstruation compared with those before intervention between the two groups(P＜0.05);the VAS scores of the observation group during the 1st,2nd and 3rd menstruation were significantly lower than those of the control group(P＜0.05).There were significant differences in dysmenorrhea symptom scores during the 1st,2nd and 3rd menstruation with those before intervention between the two groups(P＜0.05);the dysmenorrhea symptom score of the observation group during the 1st,2nd and 3rd menstruation was significantly lower than those of the control group(P＜0.05).The total effective rate of the observation group was significantly higher than that of the control group(P＜0.05).There were significant differences in PGE2 and E2 levels during the 1st,2nd and 3rd menstruation with those before intervention between the two groups(P＜0.05).There were significant differ-ences in PGE2 and E2 levels between the two groups during the 1st,2nd and 3rd menstruation(P＜0.05).There were no significant differences in menstrual cycle,menstrual duration and menstrual volume between the two groups(P＞0.05).Conclusion Balanced cupping intervention has a good effect and high safety in the treatment of patients with primary dysmenorrhea differentiated as type of qi stagnation and blood stasis.

ObjectiveTo observe the effects of the kidney-tonifying and blood-activating prescription on the Wnt/β-catenin signaling pathway and uterine spiral artery remodeling in a mouse model of recurrent miscarriage and to explore its underlying mechanism. MethodA mouse model of normal pregnancy was established by mating CBA/J mice with BALB/c mice. A mouse model of recurrent miscarriage was established by mating CBA/J mice with DBA/2 mice. The modeled mice of recurrent miscarriage were randomized into model， dydrogesterone， and low- and high-dose Chinese medicine groups. The mice in normal pregnancy were used as the control group. Each group consisted of 10 mice， and the drug administration lasted for 14 days. After the treatment， the embryo absorption rate of each group was recorded. Hematoxylin-eosin （HE） staining was employed to observe the pathological morphology of the uterine decidua， and the physiological transformation rate of spiral arteries （SPA） was evaluated. Real-time polymerase chain reaction （Real-time PCR） and Western blot were performed to determine the mRNA and protein levels， respectively， of matrix metalloproteinases （MMP）-2， MMP-9， vascular endothelial growth factor （VEGF）， and Wnt/β-catenin signaling pathway. ResultCompared with the control group， the model group presented increased embryo absorption rate （P<0.05）， decreased physiological transformation rate of uterine SPA （P<0.05）， cellular swelling， degeneration， and disordered arrangement in the uterine decidua tissue， and down-regulated mRNA and protein levels of key factors involved in SPA remodeling （MMP-2， MMP-9， VEGF） and the Wnt/β-catenin signaling pathway （Wnt2， β-catenin， Cyclin D₁， c-Myc） （P<0.05）. Compared with the model group， both the low- and high-dose Chinese medicine reduced embryo absorption rate （P<0.05）， increased SPA physiological transformation rate （P<0.05）， improved uterine decidua tissue morphology， and increased decidua vessel count. Furthermore， they up-regulated the mRNA and protein levels of MMP-2， MMP-9， VEGF， and proteins in the Wnt/β-catenin signaling pathway （P<0.05）. ConclusionRecurrent miscarriage is associated with impaired uterine spiral artery remodeling. The kidney-tonifying and blood-activating prescription can promote uterine spiral artery remodeling by activating the Wnt/β-catenin signaling pathway and promoting the expression of VEGF， MMP-2， and MMP-9， thus treating recurrent miscarriage.