Objective:To observe the effects of Xiaozhiling on the serum vascular endothelial growth factor(VEGF) level of the patients with hemangioma.Methods:10 healthy subjects (control group) and 10 hemangioma patients (treatment group) were included.The serum VEGF level were evaluated by ELISA in both groups.The treatment group was injected with Xiaozhiling once.VEGF level of the treatment group were measured before, one week after and one month(only 8 patients) after Xiaozhiling injection.Results:The serum VEGF concentrations (pg/ml) of the healty subjects, hemangioma patients without treatment, the patients 1 week after injection of Xiaozhiling and those 1 month after injection were 97.3?15.1,799.3?84.1,828.1?68.8 and 105.6?13.5 respectively. 1 week after injection tumor volume decreased by more than 25% in all the 10 patients.Conclusions:In situ injection of Xiaozhi ling may be a method for the treatment of hemangioma.

Objective To establish the quality standard of Compound Kangganling Granules. Methods TLC method was used for qualitative identification of Lonicerae Japonicae Flos and Forsythiae Fructus, and HPLC method was used for determination of chlorogenic acid in the preparation. The HPLC separation was performed on Diamonsil C18 column. The mobile phase was consisted of acetonitrile-0.4%phosphoric acid (20∶80, V/V), and the flow rate was 1.0 mL/min. Results TLC identified chlorogenic acid in Lonicerae Japonicae Flos and phillyrin in Forsythiae Fructus. The linear relationship of chlorogenic acid in the preparation measured by HPLC was A=30461C-5938.8, r=0.9998, RSD=1.01%, showing that the linear range of chlorogenic acid was 10.2-102.0μg/mL. Conclusion The method is accurate and rapid, with good stability, reliability and reproducibility, and can be used for the quality control and evaluation of the preparation.

Survivin variants specific real time quantitative RT-PCR was developed to analyze their expression in 53 paired cancer and para-cancerous tissues, and the expression of the wild-type survivin protein was detected by immunohistochemistry. The results showed that survivin mRNA and protein were expressed in gastric cancer and para-cancerous tissues. The survivin-2B was dominantly expressed in para-cancerous tissues, whereas the survivin-DeltaEx3 was more frequently detected in cancer tissues. The positive rate of survivin-2a was 100% in both cancer and para-cancerous tissues, but its relative transcript expression level was not significantly increased in cancer tissues in comparison with para-cancerous tissues. The correlation analysis revealed that the expression of survivin-2a mRNA was significantly associated with that of total survivin (r (s)=0.4178, P=0.0018), whereas inversely to that of survivin-DeltaEX3 (r (s)=-0.4506, P=0.0007). It was suggested that survivin-2a may act as an antagonist of survivin-DeltaEX3. The balance between antiapoptotic survivin iso-forms and nonantiapoptotic ones may play an important role in tumorigenesis and tumor progression. Promising value is hinted to analyze survivin and its variants in tumor early diagnosis and distinguishing malignant tumors from benign ones.
Microtubule-Associated Proteins/genetics ; Microtubule-Associated Proteins/*metabolism ; Protein Isoforms/genetics ; Protein Isoforms/metabolism ; RNA, Messenger/genetics ; RNA, Messenger/metabolism ; Stomach Neoplasms/*metabolism ; Stomach Neoplasms/pathology ; Tumor Cells, Cultured ; Tumor Markers, Biological/genetics ; Tumor Markers, Biological/*metabolism

BACKGROUND:Neurotrophin-3 is found in the repair of spinal cord injury in the role of the strongest neurotrophic factor.It can effectively promote axonal regeneration through the glial scar tissue in repairing spinal cord injury.OBJECTIVE:To construct recombinant lentiviral vectors for gene delivery of homo sapiens neurotrophin-3(hNT3),and to investigate the expression of hNT3 gene in Schwann cells after transfection.DESIGN,TIME AND SETTING:Observational experiment was performed from June 2007 to March 2008 at the Central Laboratory of Changhai Hospital.MATERIALS:Bilateral sciatic nerves were harvested from 3-day-old Sprague Dawley rats for culture and identification of Schwann cells.Three-plasmid lentivirus systems:pGC-E1-EGFP,pHelper 1.0 and pHelper 2.0 were gained from Shanghai Chemical Technology Co.,Ltd.METHODS:pGC-E1-hNT3-EGFP plasmid was constructed by double restriction enzyme digestion and ligation,and then the plasmid was transformed into E.coli DH5?.Purified pGC-E1-hNT3-EGFP plasmids from the positive clones was confirmed by PCR and sequencing.293T cells were cotransfected with lentiviral vector pGC-E1-hNT3-EGFP,pHelper 1.0 and pHelper 2.0 by Lipofectamine 2000 to produce lentivirus.2 mL recombinant virus complete culture solution was added according to multiplicity of infection=1,4,8,10,12.The titer of virus was tested according to the expression level of enhanced green fluorescent protein.The control groups were Schwann cells and Schwann cells transfected by no-loaded lentivirus.MAIN OUTCOME MEASURES:The lentiviruses were transduced to Schwann cells,and the transfection efficiency was examined by flow cytometry,the overexpression of hNT3 was determined by Real-time PCR and Western blotting.RESULTS:The exogenous gene sequence of the recombinant hNT3 was completely in accordance with that of its open reading frame in GeneBank.The titer of concentrated virus was 5?107 TU/L.After recombinant LV-hNT3 infection,Schwann cells gave off strikingly bright green fluorescence,and the transfection efficiency amounted to 85%(multiplicity of infection=10).Real-time RCR test showed that the hNT3 mRNA were highly expressed in hNT3-Schwann cells,while did not express in the control group.Western blotting showed the hNT3 expression in Schwann cells.CONCLUSION:Construction of hNT3 gene lentiviral vector can transfect Schwann cells leading to an efficient overexpression of hNT3.

Objective To investigate the effect of endothelial progenitor cells (EPCs) which complexed with neuron stem cells (NSCs) after transplanted into ischemia and reperfusion (I/R) rat model on the proliferation and apoptosis of the NSCs and vasal construction. Methods Totally 150 SD adult male rats were randomly and equally divided into 5 groups, sham-operation group, I/R group, I/R+NSCs group, I/R+EPCs group, and I/R+NSCs+EPCs group. The rats were further divided into 5 subgroups according to the time points of 3, 7, 14, 30 and 60 d after transplantation. I/R rat model was established by reversiblyligating middle cerebral artery occlusion. NSCs were derived from the hippocampus of SD rats born in 24 h and identified with Nestin staining. EPCs were obtained from the artery blood of SD rats and verified with immunocytochemical stainings of CD31, CD34 and KDR after culture. The complex of NSCs and EPCs was produced with aid of laminin. Then the complex were transplanted into the ischemia penumbra of corresponding model rat brain. The proliferation and apoptosis of NSCs, and the fomation of new vessels were observed through immunohistochemistry and double-labeled immunofluorescence staining. Results The proliferation of NSCs was increased, apoptosis cells were decreased and the new vessels were raised in the complex transplantation when compared with single NSCs or EPCs transplantation groups. Conclusion The complex of NSCs and EPCs transplantation improves the proliferation of NSCs, suppresses cell apoptosis and promotes the formation of new vessels.

Objective To explore the effect of the complex containing neuron stem cells(NSCs)and endothelial progenitor cells(EPCs)after being transplanted to ischemia and reperfusion rat model on the differentiation of neural stem cells to neuron and synapse formation.MethodsTotally 300 SD adult male rats were randomly divided into sham operation group,middle cerebral artery occlusion(MACO)model group,MACO+NSCs group,MACO+EPCs group,and MACO+complex group,and every group were further divided into 6 subgroups according to time points 1,3,7,14,30 and 60 d after operation,NSCs were isolated from the hippocampus of neonatal SD rats born in 24 h and identified with Nestin staining.EPCs were obtained from the artery blood of SD rats and verified with immunocytochemical stainings of CD31 and CD34 after culture.NSCs and EPCs were cultured together to produce the complex with aid of laminin.Then normal saline,the NSCs,EPCs,or complex(2?1010/L)were transplanted into the ischemia penumbra of corresponding model rat brain,sham control or MACO rats.The differentiation of NSCs,the expression of P38,synapsin-I,mRNA of connexin 43 were observed with double label immunofluorescence technique,immunohistochemical method,RT-PCR and Western blot analysis in ischemic penumbra.ResultsCompared with other groups,there were more double Brdu and neural special enolase positive cells in MACO+complex group with the increased expression of P38.RT-PCR and Western blot analysis indicated that MACO+complex group had significantly higher expressions of synapsin-I at mRNA and protein levels in ischemic penumbra,and so did the mRNA expression of connexin 43.ConclusionThe transplantation of the complex of NSCs and EPCs improves the NSCs differentiation to neuron,synapse formation and reconstruction of neural network.

AIM:To establish the quality standard for Nabao Capsule(Radix Astragali,Radix Angelicae sinensis,Rhizoma Corydalis,Herba Hedyotis,etc.).METHODS:Radix Angelicae sinensis,Rhizoma Corydalis,Herba Hedyotic and Pendulous Monkshood Root in Nabao Capsule were identified by TLC.Astragaloside Ⅳ was determined by HPLC.RESULTS:The characteristic identification by TLC was distinct and highly specific,the quantitative evaluation of thymol had the linear range of 1-10 ?g.The average recovery was 99.05% and RSD was 2.48%.CONCLUSION:The method is reliable,accurate and specific,and can be used for the quality control of Nabao Capsule.

BACKGROUND:Chemotherapy drugs can damage the ovarian function in women of childbearing age, and even lead to premature ovarian failure. Therefore, to improve and restore the ovarian function in patients has become an important issue. OBJECTIVE:To explore the therapeutic effect and feasibility of bone marrow mesenchymal stem cel therapy against chemotherapy-induced ovarian damage. METHODS:Rat models of chemotherapy-induced premature ovarian failure were established, and injected with PKH26-labeled bone marrow mesenchymal stem cels. At 15, 30, 45, 60 days after cel transplantation, five rats were selected respectively to detect folicle-stimulating hormone and estradiol levels, and then, the rats were kiled to take the right ovary for pathological examination. The number of ovarian folicles was detected under light microscope. At 30 days after cel transplantation, another two rats were selected to mate with male rats to observe the difference in the reproductive activity. RESULTS AND CONCLUSION:Four of 22 rats (18%) gradualy recovered their estrous cycle after cel transplantation, with the decreased folicle-stimulating hormone level and increased estradiol level. Moreover, the number of folicles was reduced. Al of these indicated that the ability to have children in rats was not damaged.These experimental findings suggest that bone marrow mesenchymal stem cels can partialy improve the ovarian function of rats under chemotherapy.

Objective To investigate the infection sites,the offending species,diagnosis and prognosis of deep fungal infections (DFI) in patients with systemic lupus erythcmatosus (SLE).Methods Fifty-one patients with fungal infections in 1466 SLE patients admitted to Peking Union Medical College (PUMC) Hospital from 2000 to 2006 were reviewed retrospectively.Results Candida albicans was ranked the first pathogen,followed by ncoformans and Aspergillus species.The infection sites were lungs,cerebral meninges and blood in the order of prevalence.The overall mortality was 20%(10/51).Aspergillasis carried the highest mortality which couht be as high as 80%.Hypoproteinemia,multiple focus of fungal infections,Aspergillasis and fungemia might he the independent risk factor for mortality.Conclusion Candida albicans is the most frequent species of fungal infections in SLE patients.Iungs are the most prevalent location of infection.Earlier diagnosis is important.Special attention should be paid to aspergillasis.

Objective To analyze the clinical effect of debridement in the treatment of Kashin-Beck disease under an ankle arthroscopy.Methods Totally 40 patients with ankle Kashin-Beck disease who had underwent surgery in the Fifth Hospital of Daqing from January 2015 to October 2016 were selected as the research subjects.All patients were treated with cleaning treatment under arthroscopy,evaluated via the visual analogue scale method,and compared pain before and 6,12 month after the treatment.Walking function and ankle motion of the patients were evaluated via an ankle-scoring system,and the quality of life of patients was investigated before and after the treatment through a questionnaire survey.Results The visual analogue scale of before and 6,12 months after the treatment was (6.2 ± 1.0),(2.3 ± 1.2) and (1.6 ± 0.4) scores,respectively;ankle foot score (53.3 ± 6.8),(86.6 ± 5.1) and (88.7 ± 6.2) scores,respectively;ankle activity score (15.9 ± 4.1),(36.9 ± 4.9) and (41.8 ± 6.4) scores,respectively;and life quality score (53.1 ± 4.0),(75.4 ± 6.6) and (88.7 ± 10.5) scores,respectively.The differences between groups were statistically significant (F =2.1,2.6,2.3,2.5,P ＜ 0.05).Visual analogue scale of 6 and 12 months after the treatment was significantly lower than that before the treatment,ankle foot score and ankle activity score and life quality score were significantly higher than those before the treatment (P ＜ 0.05).Conclusion The debridement under an ankle arthroscopy can effectively reduce the pain of patients with Kashin-Beck disease,and improve the walking function of the patients obviously,significantly improve the patients' life quality,and the method should be widely applied in clinical treatment.