Objective To study the method of extracting the gypenoside from crude Herba Gynostemmae Pentaphylli solution.Methods The gypenoside was extracted from crude Herba Gynostemmae Pentaphylli solution by foaming separation.The optimum conditions were obtained through calculating the recovery rate and enrichment ratio.Results Under the optimum conditions:the concentration of the gypenoside in crude Herba Gynostemmae Pentaphylli solution was 3.21 ?g/mL,the gas flow rate was 30 mL/min,the volume of the solution was 110 mL,and pH value was 9.0 as well,the recovery rate of gypenoside was 49.20% and the enrichment ratio could reach 4.511.Conclusion Foam separation is a simple and effective method of extracting gypenoside from crude Herba Gynostemmae Pentaphylli solution.

Objective To retrospectively analyze our experience of embolization of ruptured intracranial aneurysms during the period of cerebral vasospasm (CVS). Methods Thirty-seven patients with ruptured intracranial aneurysms were embolized with electrolytic detachable coils during the period of CVS (days 4 to 14). Group A included the 14 patients with angiographic CVS and group B included 23 patients without angiographic CVS. All except 2 patients were transferred to our department during the CVS period. Glasgow outcome score (GOS) was evaluated 3 months after the treatment. Results Twelve patients in group A successfully received the aneurysm embolization and treatment of the CVS with intraarterial papaverine injection and balloon angioplasty. GOS in 3 months was good recovery in 7 patients, moderate disability in 2, severe disability in 1, and dead in 2, respectively. Embolization failed in 2 patients because the microcatheters could't pass the spasmodic parent arteries. All the aneurysms in group B were successfully embolized. GOS were good recovery in 18 patients, moderate disability in 2, severe disability in 2, and dead in 1, respectively. There was no intraprocedural aneurysmal rupture but with 2 thromboembolic events. No rebleeding occurred during the mean 11 months follow-up. Conclusions The so-called “period of CVS” isn′t always associated with CVS in angiograpy. Embolization of ruptured intracranial aneurysms during the period of pure CVS doesn′t carry an increased risk. Both the aneurysms and CVS can be treated during the single procedure. It can reduce the rebleeding rate in hospital and improve the prognosis of the patients with CVS.

Blood-brain barrier(BBB)is mainly composed of brain microvascular endothelial cells(BMECs).According to the special cell membrane transport system and the extensive tight junctions,the brain microvascular endothelial cells play an important role in the CNS.Also,there is a close relationship between BMECs and the cerebrovascular diseases,such as cerebral ischemia,cerebral edema,and so on.Therefore,the culture of brain microvascular endothelial cells provides a new tool for studying the cerebrovascular diseases in vitro.The researches in recent years have been reviewed in this test,approaching the cultivation of the rat brain microvascular endothelial cell from the segregation,purification and culture in turns,in order to provide a foundation for the studies of the brain microvascular endothelial cell;in vitro.

Objective To explore the clinical treatment of patients with stage Ⅲ carcinoma of colon and rectum by tegafur chemotherapeutic drugs combined with standardized adjuvant therapy. Methods The data of 60 cases of carcinoma of colon and rectum treated in Wenzhou Central hospital from January 2013 to December 2015 were collected. The patients were divided into three groups, non standardized adjuvant treatment group of 18 cases, standardized adjuvant treatment group of 15 cases and tegafur drug combined with standardized treatment group of 27 cases. Recorded and compared the survival rate of the three groups within three years, the statistics of each group of disease-free survival (DFS) and overall survival rate (OS). According to the duration of treatment group, the use of oxaliplatin combined with 5-fluorouracil and leucovorin (FOLFOX) treatment, analyzed and recorded OS and DFS. Follow-up examinations were performed in patients who did not receive adjuvant therapy. Recorded and compared the OS and DFS of each group. Results Tegafur drug combined with standardized treatment group DFS and OS were significantly higher than the other two groups, the difference was statistically significant (P<0.05). In addition, when treated with FOLFOX regimen, the DFS and OS of group Ⅲ were significantly higher than those of groupⅠand groupⅡ, the difference was statistically significant (P<0.05). In the follow-up survey of three groups, the rates of recurrence and death in patients treated with tegafur drug combined with standardized treatment group were significantly lower than those in the non standardized adjuvant treatment group and the standardized adjuvant treatment group. Conclusion Tegafur combined with adjuvant therapy can significantly improve the standardization of the rate of carcinoma of colon and rectum and the rate of disease-free survival, it is suggested that doctors use chemotherapy combined with auxiliary standardized therapy, can significantly improve the survival of patients, and reduce the risk of disease recurrence rate.

Objective:To explore effect of early rehabilitation therapy on improving prognosis and quality of life (QOL) in patients with acute myocardial infarction (AMI) .Methods :A total of 120 AMI patients treated in our hospital were selected .According to random number table ,they were randomly and equally divided into routine treatment group (n=60 ,received routine treatment) and rehabilitation group (n=60 ,received routine treatment combined rehabilitation therapy ) .Clinical symptoms ,myocardial enzymes ,cardiac function indexes ,complications , QOL ,length of hospital stay and hospitalization cost were statistically analyzed and compared between two groups . Results :Compared with routine treatment group after treatment ,there were significant reductions in incidence rates of dizziness ,low back pain ,leg weakness ,constipation ,palpitations (P<0.05~ <0.01) ,abdominal distension and complications (36.7% vs .5.0% );significant rise in Barthel index [ (61.9 ± 8.7) scores vs .(86.4 ± 6.9) scores] , significant reductions in length of hospital stay [(13.8 ± 3.2) d vs .(5.9 ± 2.6) d] and hospitalization cost [(13600 ± 1450) RMB vs .(8600 ± 1240) RMB] in rehabilitation group ,P<0.05~ <0.01 .Conclusion:Early rehabilitation therapy can significantly improve prognosis and quality of life ,shorten length of hospital stay , lower hospitalization cost in patients with acute myocardial infarction ,which is worth clinical extending .

Objective To investigate the clinical value of of procalcitonin detection in neonatal necrotizing enterocolitis (NEC).Methods Sixty cases eligible NEC infants were randomly divided into observation group and control group(n=30 each group).All the infants were divided into Ⅰ grade group(19cases),Ⅱ-Ⅲ grade group(41 cases) according to NEC classification.All infants received PCT detection after NEC diagnosis.The observation group received antibiotic treatment base on the serum PCT levels.The control group received antibiotic treatment according to treatment routine and physician's clinical experience.Results PCT increased in 28 (46.7％) of the 60 NEC infants.In the Ⅰ grade group and Ⅱ-m grade group,the proportion of PCT increased cases were 26.3％,61.3％ respectively,and the PCTmax in PCT positive cases were (0.81±0.25) μg/L,(1.76±0.89) μg/L respectively,the differences were signifiacnt(x2 =4.627,t=-2.608,P＜0.05).The time of antibiotic usage in observation group was significantly less than that in control group ((4.9±3.0) d vs.(8.6±2.5) d,t=-3.645,P＜0.01).The fasting time and the cure time of the two groups had no significant difference (P＞ 0.05).Conclusion PCT detection can help to understand the cause and severity of NEC.Using antibiotics according to PCT levels can reduce the time of antibiotic usage without affecting the effect of the NEC treatment.

Objective Autologous hematopoietic stem cell transplantation (Auto-HSCT) has been widely used in hematological malignancies.To mobilize and harvest sufficient number of peripheral CD_(34)~+ cells is one of key issues for auto-HSCT. Peripheral CD_(34)~+ cell numeration has been used as an indicator for apheresis while we mostly rely on the peripheral WBC or MNC count. In this study, we try to evaluate the association of peripheral CD_(34)~+ count to the CD_(34)~+ cells number in the apheresis product and to find out a potential threshold. Methods From Jan 2007 to Dec 2009, a total of 57 apherosis for auto-HSCT were analysed. All patients were mobilized by cyclophophamide (CTX) plus G-CSF(5-10μg/kg) regimen. The apheresis were performed with COBE SPECTRA VERSION 6 and CD_(34)~+ count of both peripheral and apheresis products were analysed by flow cytometry. Results The median number of MNC in apheresis products was 4.6(0.3-10.5)×10~8/kg with median CD_(34)~+ cells at 2.4(0.16-34.9)×10~6/kg. The peripheral CD_(34)~+ count was the only parameter associated with the MNC and CD_(34)~+ cell numbers in the apheresis products while the WBC number was irrelevant to the results of apheresis. Our data showed that when the peripheral CD_(34)~+ count reach 15/μl, the efficacy of a single apheresis significantly improved with 81 % and 60 % reached 1 and 2×10~6 CD_(34)~+ cells/kg respectively and the total number of MNC and CD_(34)~+ cells were significantly superior to apheresis with peripheral CD_(34)~+ cells <15/μl, thus indicated that CD_(34)~+ ≥15 /μl can be used as the threshold for apheresis. Furthermore, the ROC analysis demonstrated that CD_(34)~+ cells ≥25(26.5-28.6) /μl is the best indicator level for a successful single apheresis. Conclusion Our study clearly showed that peripheral CD_(34)~+ cell count is a key indicator of apherosis. CD_(34)~+ cells at 15/μl can be used as the threshold to start apheresis in the clinical setting.

ObjectiveTo investigate the effect of tacrolimus (FK506) on macrophage accumulation,proliferation and activation in the kidney of early diabetic rats and to explore its possible mechanism of renal protection.Methods Rats were randomly divided into control,model and tacrolimus groups.Diabetic model rats were induced with intraperitoneal injection of streptozotocin.Tacrolimus(0.5 or 1.0 mg·kg-1 ·d-1) was orally administered once a day for 4 weeks.Kidney weight index(KWI),24-h urinary albumin excretion rate(UAER) and creatinine clearance rate(Ccr) were measured.Kidney pathology was observed by light microscopy.ED-1,PCNAandiNOSpositivemacrophagesweredetectedbysingleanddoublestainingof immunohistochemistry.Results KWI increased in model group and was significantly reduced by tacrolimus treatment with 1.0 mg·kg-1 ·d-1 (P＜0.05).UAER elevated in model group and was markedly attenuated by tacrolimus treatment with 0.5 and 1.0 mg·kg-1 ·d-1 (P＜0.05).Elevated glomerular volume of model rats was significantly decreased by tacrolimus treatment with 0.5 and 1.0 mg·kg-1·d-1 (P＜0.05),and increased indices of tubulointerstitial injury were only ameliorated by 1.0 mg·kg-1·d-1 tacrolimus(P＜0.01).Marked accumulation of ED-1+ cells in diabetic kidney was found,which was not inhibited by tacrolimus treatment with 0.5 and 1.0 mg·kg-1·d-1.ED-1PCNA+ cells and ED-1+ iNOS+ cells were significantly elevated in kidneys of model group,while they were significantly inhibited by tacrohmus treatment with 0.5 and 1.0 mg·kg-1·d-1 (P＜0.01).Conclusion Tacrolimus can ameliorate early renal injury of diabetic rats and its mechanism may be partly associated with the suppression of increased macrophages activation.

BACKGROUND:The mechanism of differentiation and proliferation of bone marrow-derived mesenchymal stem cells remains unclear. In addition, issues such as how signal pathways such as Ca2+and bone marrow-derived mesenchymal stem cellproliferation and differentiation signals form complex signal network remain poorly understood.
OBJECTIVE:To investigate the effect of Ca2+in the induced differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes.
METHODS:Bone marrow-derived mesenchymal stem cells were isolated from rat bone marrow using whone bone marrow adherence method, purified, amplified, and induced with hepatocyte growth factor. [Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells and control bone marrow-derived mesenchymal stem cells were detected with flow cytometry. Bone marrow-derived mesenchymal stem cells induced with hepatocyte growth factor were mixed with nimodipine of different concentration, and cells were divided into three groups:hepatocyte growth factor+nimodipine 10 mg/L, 50 or 100 mg/L groups. cellgrowth was observed with inverted phase contrast microscope and alpha 1-antitrypsin expression of the cells was confirmed by immunocytochemistry. The calcineurin M and the activation of extracellular signal regulated kinase pathway was detected by reverse transcription-PCR and western blotting, respectively.
RESULTS AND CONCLUSION:[Ca2+]i in the directional differentiated bone marrow-derived mesenchymal stem cells was higher than in the control group (P<0.05). After addition of a larger dose of nimodipine, no differentiation of cells was obeserved and growth of bone marrow-derived mesenchymal stem cells was getting worse. There were few alpha 1-antitrypsin positive cells in the nimodipine groups. Calcineurin Mexpression was significantly increased in directional differentiated bone marrow-derived mesenchymal stem cells and smal dose of nimodipine than the controls (P<0.05). However, no significant difference was found among middle, high dose nimodipine and control groups (P>0.05). These findings indicate that Ca2+could participate in the differentiation of bone marrow-derived mesenchymal stem cells into hepatocytes incuded with cytokines, and also maintain the survival and proliferation of bone marrow-derived mesenchymal stem cells.

ObjectiveTo explore the practicability of combined surgical treatment for the patients with Hepatocellular Carcinoma(HCC) complicated with Portal Hypertension(PH).MethodsThe clinical data of 43 patients withHCCcomplicatedwithsignificantesophagealvaricesandhyperspleniawereanalyzed retrospectively.All patients were diagnosed with esophageal varices and hypersplenia of different degrees, 14 of them had severe esophageal varices, the other 29 patients had moderate esophageal varices.Firstly all patients underwent splenectomy, 18 of them had selective portoazygous devascularization in addition.After that the patients received hepatectomys ,with local excision or segmentectomy as the major operation methods.Results All patients received hepatectomy combined with splenectomy treatment.There was no perioperative mortality, but 12 patients had complications after the surgery.Over seventy percents patients(33/43)accepted regular followup.Among these 33 patients,the 1-year and 3-year survival rates were 90.9％ (30/33) and 63.6％ (21/33)respectively.Eleven patients died of tumor recurrence.Four patients had upper postoperatively gastrointestinal hemorrhage.ConclusionHepatectomy combined with splenectomy didn't increase the mortality in the patients with HCC complicated with PH.The combined surgery might reduce the incidence of severe complications, such as postoperative bleeding.We believe the combined surgical treatment is feasible for this group of the patients.