2.Inhibition of PI3K/PKB signal pathway improves chemotherapeutic effect on gastric carcinoma cell lines
Xia XIE ; Qing GAO ; Yanli WANG ; Chun TIAN
Chinese Pharmacological Bulletin 2003;0(12):-
Aim To observe whether the inhibition of PI3K/PKB signal pathway by LY294002[PI3K pathway inhibitor] could improve the sensitivities of human gastric carcinoma cell line SGC7901 and SGC7901/VCR to anti-cancer drugs.Methods The sensitivities of SGC7901 and SGC7901/VCR to chemotherapeutic drug VCR were detected by MTT.The MDR1 and XIAP mRNA expression levels were evaluated by semi-quantitative RT-PCR,and their protein levels were detected by immunocytochemistry.The PKB and phospho-PKB protein levels were detected by Western blot and the apoptosis ratio was detected by flow cytometry.Results 2?10-5mol?L-1LY294002 enhanced the sensitivities of SGC7901 and SGC7901/VCR cells to VCR(P
3.Fusion of human orphan G protein-coupled receptors GPR45, GPR85 or GPR174 with Gi1α and their expression in insect Sf9 cells
Ming-li, PENG ; Chun-guang, HAN ; Zhi-qing, GAO ; Qiong, WANG ; Yue, GAO ; Yong-xue, LIU
Bulletin of The Academy of Military Medical Sciences 2010;34(1):8-11
Objective To obtain the fusion genes of several human orphan G protein coupled receptors (oGPCRs) with Gi1α subtype of G protein and their expression system. Methods The whole open reading frames of GPR45, GPR85, GPR174 and Gilα were cloned by RT-PCR from HepG2 cDNA separately,and the corresponding fusion genes were amplified by overlap extension PCR. Then, the fusion genes-containing pBacmids were successfully constructed with the Bac-to-Bac baculovirus expression system indicated by specific transposition and virus recombination. The insect Sf9 cells were transfected with pBacmid-oGPCRs-Gi1α, and the supernatant containing recombinant virus was harvested. With the supernatant, insect Sf9 cells were infected under an optimized condition (MOI=5, infection time=72 h) and the fusion proteins were prepared and detected by Western blotting.Results The three fusion genes of GPCR45, GPR85 or GPR174 with Gi1α were obtained. The corresponding fusion proteins could be properly prepared in Sf9 cells.Conclusion Human oGPCRs could be fused with Gilα, and the fusion genes could be expressed using the Bac-to-Bac baculovirus expression system in insect Sf9 cells.
4.The clinical application of procalcitonin, leukocyte count and C-reactive protein in elderly patients with infection
Pei WU ; Chun SHAN ; Qun ZHANG ; Yinfang XU ; Yanping QING ; Chao GAO ; Ruixia SU
Chinese Journal of Geriatrics 2012;31(10):833-836
Objective To analyze and compare the clinical application values of procalcitonin (PCT),leukocyte count (WBC) and C-reactive protein(CRP) in elder patients with infection.Methods In patients(age≥ 65 yrs,axillary temperature >38.0℃)with infection or suspected infection,PCT,WBC,CRP and other bacteriological examination were performed.The electronic medical records from the HIS system of our hospital were analyzed retrospectively in combination with medical history.Results Of the enrolled 219 patients,65 ones were in control group,48 ones SIRS,51 ones sepsis and 55 ones MODS.There was a positive correlation between the level of serum PCT and the infection degree.The Spearman correlation coefficient was 0.706 (95%CI:0.616-0.797,P=0.000).Based on the highest Youden index (sensitivity+specificity-1),the best cutoff point of diagnosis for PCT was >0.341 μg/L (sensitivity 84.5%,specificity 55.8%),a analysis of receiver operating characteristic(ROC) curve about PCT,WBC and CRP was carried.Area under the curve (AUC) of PCT to controlled infection was 0.916 (95%CI:0.864-0.967,P=0.000).Based on the highest Youden index (sensitivity+ specificity-1),the judging threshold of PCT to infection controlled or not was 0.73 μg/L (sensitivity 84.6%,specificity 88.0%).PCT level after treatment >0.73 μg/L showed the uncontrolled infection,< 0.73 μg/L controlled.Conclusions PCT has a higher specificity for elder patients with infection.The variation of PCT level can guide the application of antibiotics,avoid abuse and decrease the occurrence of drug-resistant bacteria.
5.Effect of electro-acupuncture stimulation of Ximen (PC4) and Neiguan (PC6) on remifentanil-induced breakthrough pain following thoracal esophagectomy.
Yan-Hu, XIE ; Xiao-Qing, CHAI ; Yue-Lan, WANG ; Yan-Chun, GAO ; Jun, MA
Journal of Huazhong University of Science and Technology (Medical Sciences) 2014;34(4):569-74
The clinical analgesic effect of electro-acupuncture (EA) stimulation (EAS) on breakthrough pain induced by remifentanil in patients undergoing radical thoracic esophagectomy, and the mechanisms were assessed. Sixty patients (ASAIII) scheduled for elective radical esophagectomy were randomized into three groups: group A (control) receiving a general anesthesia only; group B (sham) given EA needles at PC4 (Ximen) and PC6 (Neiguan) but no stimulation; and group C (EAS) electrically given EAS of the ipsilateral PC4 and PC6 throughout the surgery. The EAS consisting of a disperse-dense wave with a low frequency of 2 Hz and a high frequency of 20 Hz, was performed 30 min prior to induction of general anesthesia and continued through the surgery. At the emergence, sufentanil infusion was given for postoperative analgesia with loading dose of 7.5 μg, followed by a continuous infusion of 2.25 μg/h. The patient self-administration of sufentanil was 0.75 μg with a lockout of 15 min as needed. Additional breakthrough pain was treated with dezocine (5 mg) intravenously at the patient's request. Blood samples were collected before (T1), 2 h (T2), 24 h (T3), and 48 h (T4) after operation to measure the plasma β-EP, PGE2, and 5-HT. The operative time, the total dose of sufentanil and the dose of self-administration, and the rescue doses of dezocine were recorded. Visual Analogue Scale (VAS) scores at 2, 12, 24 and 48 h postoperatively and the incidence of apnea and severe hypotension were recorded. The results showed that the gender, age, weight, operative time and remifentanil consumption were comparable among 3 groups. Patients in EAS group had the lowest VAS scores postoperatively among the three groups (P<0.05). The total dose of sufentanil was 115±6.0 μg in EAS group, significantly lower than that in control (134.3±5.9 μg) and sham (133.5±7.0 μg) groups. Similarly, the rescue dose of dezocine was the least in EAS group (P<0.05) among the three groups. Plasma β-EP levels in EAS group at T3 (176.90±45.73) and T4 (162.96±35.00 pg/mL) were significantly higher than those in control (132.33±36.75 and 128.79±41.24 pg/mL) and sham (136.56±45.80 and 129.85±36.14 pg/mL) groups, P<0.05 for all. EAS could decrease the release of PGE2. Plasma PGE2 levels in EAS group at T2 and T3 (41±5 and 40±5 pg/mL respectively) were significantly lower than those in control (64±5 and 62±7 pg/mL) and sham (66±6 and 62±6 pg/mL) groups. Plasma 5-HT levels in EAS group at T2 (133.66±40.85) and T3 (154.66±52.49 ng/mL) were significantly lower than those in control (168.33±56.94 and 225.28±82.03) and sham (164.54±47.53 and 217.74±76.45 ng/mL) groups. For intra-group comparison, plasma 5-HT and PGE2 levels in control and sham groups at T2 and T3, and β-EP in EAS group at T3 and T4 were significantly higher than those at T1 (P<0.05); PGE2 and 5-HT levels in EAS group showed no significant difference among the different time points (P>0.05). No apnea or severe hypotension was observed in any group. It was concluded that intraoperative ipsilateral EAS at PC4 and PC6 provides effective postoperative analgesia for patients undergoing radical esophagectomy with remifentanil anesthesia and significantly decrease requirement for parental narcotics. The underlying mechanism may be related to stimulation of the release of endogenous β-EP and inhibition of inflammatory mediators (5-HT and PGE2).
6.Effect of Runing II on expression of vascular endothelial growth factor in transplanted tumor of mammary cancer MA-891 in TA2 mice.
Xue-qing WU ; Shang-pu GAO ; Ming-chun MU
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(3):251-253
OBJECTIVETo study the effect of Runing II on expression of vascular endothelial growth factor (VEGF) in transplanted tumor of mammary cancer MA-891 in TA2 mice.
METHODSThe MA-981 mice mammary cancer cell cultivated in vivo was inoculated into the right axilla subcutaneously of TA2 mice to establish the transplanted tumor model, which were treated with Runing II.
RESULTSRuning II could inhibit the growth of transplanted tumor and the occurrence of lung metastasis (P < 0.05), reduce the expression of VEGF protein and mRNA in tumor tissue (P < 0.05).
CONCLUSIONRuning II could reduce the expression of VEGF protein and mRNA, hence to inhibit the growth of tumor and lung metastasis.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Female ; Mammary Neoplasms, Experimental ; metabolism ; Mice ; Neoplasm Transplantation ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vascular Endothelial Growth Factor A ; biosynthesis ; genetics
7.The effect of partial body-weight supported treadmill training on hemiplegia patients caused by with cerebral infarction
Cui-Huan PAN ; Ai-Hua LUO ; En XU ; Wen-Wei WANG ; Qing-Chun GAO ; Tong YE ; Yi HUANG ;
Chinese Journal of Physical Medicine and Rehabilitation 2003;0(10):-
Objective To investigate the effect of partial body-weight supported treadmill training ( PBW- STT) on function of lower limbs, walk function, ADL performance and quality of life of hemiplegic patient induced by cerebral infarction. Methods A total of 132 cerebral infarction patients were divided into a control group (n = 69) and a training group( n = 63) randomly. Both groups accepted routine rehabilitation therapy, and the training group accepted PBWSTT at the same time in addition. Both groups were evaluated with regard to their walking ability, func- tion of lower limbs, ADL performance and their quality of life by using Functional Ambulation Category (FAC) , Fugl-Meyer assessment (FMA) , Barthel index (BI) and SF-36 before and after rehabilitation treatment. Results The function of lower limb, walking ability, ADL performance and the quality of life of both groups were improved significantly after treatment, and those in the training group were improved to a significantly greater extent than those in the control group ( P
8.Gene transfection of NGF and/or Noggin affecting the differentiation of bone marrow-derived mesenchymal stem cells
Jie DING ; Shan GAO ; Yon CHENG ; Chun-Sheng ZHANG ; Xi-Qing HU
Chinese Journal of Neuromedicine 2010;9(6):553-557
Objective To explore the feasibility of exogenous NGF and/or Noggin gene transfecting into the bone marrow-derived mesenchymal stem cells (BMSCs) and observe the differentiation of BMSCs modified by NGF and/or Noggin.Methods BMSCs were isolated from SD rat and purified by adherent method and these cells were identified by their phenotypical properties and their abilities of differentiating into adipocytes.Ad-GFP-NGF and/or Ad-GFP-Noggin were transfected into BMSCs.The protein expressions of NGF and/or Noggin were detected using immunocytochemistry and Western blotting.The differentiations of gene modified BMSCs were observed by immunohistochemistry.Result The cells selected by adherent method had basic phenotypical properties of BMSCs and could differentiate into adipocytes.BMSCs without transfection and those transfected with Ad-GFP expressed low level of NGF without Noggin expression.All gene modified BMSCs could express NGF and/or Noggin.After transfection, BMSCs could differentiate into cells having neuronal morphology and expressing NF (H).The combined transfection group had the highest ratio of NF(H)+ cells among all the groups.Conclusion BMSCs can be isolated and purified from rat bone marrow by adherent method.Ad-GFP-NGF and/or Ad-GFP-Noggin can transfect BMSCs safely and the transfected cells can express those proteins persistently and efficiently.NGF and Noggin can induce BMSCs into neuron-like cells in vitro and when they exist simultaneously,the differentiation is further enhanced.
9.Perfusion strategies of extracorporeal circulation for robotically assisted cardiac surgery.
Jia-Li WANG ; Jia-Chun LI ; Jia-Chun LI ; Chang-Qing GAO ; Tao ZHANG
Journal of Southern Medical University 2009;29(11):2300-2301
OBJECTIVETo discuss the management of extracorporeal circulation (ECC) for robotically assisted cardiac surgery.
METHODSA total of 71 patients underwent cardiac surgeries using da Vinci S robotic surgical system, including 49 undergoing atrial septal defect repair (ASDR), 13 undergoing mitral valvuloplasty (MVP), 8 receiving left atrial myxoma excision and 1 having right atrial myxoma excision. For most of the patients, ECC was established through the femoral arterial cannula, femoral venous cannula and right internal jugular venous cannula, with only 1 patient undergoing MVP with femorai arterial cannula and femoral venous cannula. In all the cases, vacuum-assisted venous drainage (VAVD), continuous blood gas monitoring and ultrafiltration were used during ECC. Myocardial protection was managed with 4:1 blood cardioplegic solution (61 cases) or HTK solution (10 cases receiving MVP).
RESULTSThe mean ECC time and aortic cross-clamp time was 116.1-/+35.5 min (range 45-206 min) and 58.5-/+23.6 min (range 27-132 min) in these patients, respectively. During ECC, the mean urine volume was 825.5-/+447.3 ml (200-1900 ml) and ultrafiltration volume was 2571.1-/+885.9 ml (800-4800 ml) with subzero-balanced (58 cases) or zero-balanced (9 cases) total fluid balance. The mean postoperative intubation time was 10.1-/+5.3 h (7-17 h) and drainage volume within 24 h postoperatively was 217.5-/+167.8 ml (80-680 ml).
CONCLUSIONThe establishment of ECC system through peripheral vessels, certain learning curve of perfusion technique and close communication between the surgical team are the key points of ECC for totally robotically assisted cardiac surgery, and VAVD and continuous blood gas monitoring are essential during ECC.
Adolescent ; Adult ; Aged ; Extracorporeal Circulation ; Female ; Heart Septal Defects, Atrial ; surgery ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Mitral Valve ; surgery ; Robotics ; Young Adult
10.Effect of astragalus polysaccharides on the proliferation and ultrastructure of dog bone marrow stem cells induced into osteoblasts in vitro.
Chun-jiao XU ; Xin-chun JIAN ; Feng GUO ; Qing-ping GAO ; Jie-ying PENG ; Xi-ping XU
West China Journal of Stomatology 2007;25(5):432-436
OBJECTIVETo observe the growth and osteogenic property of cultured dog bone marrow stem cells (BMSCs) by investigating the effects of astragalus polysaccharides (APS) on the proliferation and ultrastructure of BMSCs into osteoblasts in vitro.
METHODSBMSCs osteogenic property was detected by improved Wright-Giemsa, Gomori and alizarin dyeing method. The proliferation and differentiation of the induced BMSCs with APS in different concentration and time were detected by MTT assay and the morphologic change of the induced BMSCs was observed by transmission electron microscope (TEM).
RESULTSBMSCs osteogenic property was detected with Wright-Giemsa deep-bluing, Gomori method blacking and with more mineral nodules alizarin dyeing method carmining. APS with concentration of 0.005 mg/mL can promote the proliferation of the induced BMSCs in short-term culture (1th, 3th day) and 50 mg/mL can decrease the effect through long-term culture (5th day). Observed by TEM (5th day), the number of mitochondria, rough endoplasmic reticulum increased and the extracellular matrix was excreted more in the induced BMSCs by APS with concentration of 0.005 mg/mL. However, not only the number of mitochondria, rough endoplasmic reticulum reduced but also the structure was swollen, degenerative, membrance damaged in the induced BMSCs by APS with concentration of 50 mg/mL.
CONCLUSIONAPS with lower concentration in short-term culture may promote BMSCs proliferation and differentiation.
Animals ; Bone Marrow Cells ; Cell Differentiation ; Dogs ; In Vitro Techniques ; Mesenchymal Stromal Cells ; Osteoblasts ; Polysaccharides