1.Anti-integrin β1 monoclonal antibody P5D2 inhibits invasion of human cervical squamous carcinoma HCE1 multicellular spheroids into umbilical vein endo-thelium cells
Xin ZHANG ; Yilin WU ; Fengying LIU
Chinese Journal of Cancer Biotherapy 2010;17(1):40-45
Objective: To probe into the relationship between integrin β1 expression in human cervical carcinoma HCE1 multicellular spheroids (HCE1/MCS) and their invasion into human umbilical vein endothelium cells (HUVEC), so as to assess the inhibitory effect of anti-integrin β1 monoclonal antibody P5D2 on the invasion of HCE1/MCS into HUVEC. Methods: A model of HUVEC monolayer invaded by HCE1/MCS was established (in brief, the HCE1 invading model). Morphology changes of the HCE1 invading model were observed under inverted microscope and analyzed by Motic Med System after P5D2 treatment. The expressions of β1 integrin on HCE1 monolayer cells, HCE1/MCS, HVUEC monolayer cells, and P5D2-treated HCE1 invading models were measured by immunocytochemistry SABC assay. Results: A HCE1 invading model was successfully established. The HCE1/MCS proliferated rapidly after culture, and on the 7th day the invading area of HCE1/MCS was 40.42 folds larger than the original HCE1/MCS. Invading areas of P5D2-treated HCE1/MCS were significantly smaller than that of the control group after 1, 4, 7 day (P<0.05, or P<0.01), with the invading area after 7 days reduced by (84.68±0.08) % compared with the control group. Integrin β1 expression in HCE1/MCS was significantly higher than that in HCE1 monolayer cells (P<0.01), and the expression was negative in HUVEC. Integrin β1 expression in P5D2-treated HCE1/MCS was significantly lower than that the in untreated HCE1/MCS (P<0.01). Conclusion: Upregulated expression of integrin β1 in HCE1/MCS and HCE1 invading model may be associated with their enhanced adhesion and invasion abilities. Anti-integrin β1 monoclonal antibody P5D2 can partially block the invasion of HCE1/MCS into HUVEC.
2.The significance and expression of Integrin α2β1 in serum of osteosarcoma patients
Xiaozhou LIU ; Sujia WU ; Xin SHI
Chinese Journal of Orthopaedics 2011;31(6):681-685
Objective To evaluate the expression of Integrin α2β1 in serum of osteosarcoma patients and to investigate the effect of Integrin α2β1 on metastasis of osteosarcoma. Methods Sandwich enzymelinked immunosorbent assay was applied to test Integrin α2β1 in serum of 26 healthy controls (male 16, female 10; age 18-32, average age 23.54±3.82) and 43 osteosarcoma patients (male 28, female 15;age 8-47,average age 18.42±9.10)before and after the surgery and the chemotherapy. The patients included osteoprogenitor cells type in 24 cases, fibroblast type in 9 cases, chondroblast type in 7 cases, and other types in 3cases; with Enneking stage ⅡA in 13 cases, stage ⅡB in 26 cases, stage Ⅲ in 4 cases. Among them, 22 patients receive the neo-adjuvant chemotherapy. After that, we compared the data of different groups. At the same time, in accordance with tumor size, position, clinical stage, histology grade and metastasis situation to categorize groups compared among the groups of serum Integrin α2β1 differences in order to explore its relationship with patients' prognosis and metastasis. Results The Integrin α2β1 of osteosarcoma patients in blood before the surgery was expressed highly. It was significantly higher than the control group. The expression before surgery had significant correlation with osteosarcoma Enneking clinical stages, tumor size and metastasis. And it was independent of tumor position and Dahlin histology grade. The expression of Integrin α2β1 in blood of the osteosarcoma patients after limb salvage surgery decreased significantly. And the expression of Integrin α2β1 in blood of patients receiving new adjuvant chemotherapy was significant differences before and after the chemotherapy. Conclusion The expression of Integrin α2β1 has positive correlation with the occurrence, development and metastasis of the osteosarcoma. It is probably a new biological indicator for diagnosis and estimating prognosis of osteosarcoma. Simultaneously surgery and new adjuvant chemotherapy are still effective methods to treat osteosarcoma.
3.Anatomical observation of the retromandibular vein by mandibular angle ostectomy
Weihua LIU ; Shaoping WU ; Xin LI
Chinese Journal of Tissue Engineering Research 2010;14(48):9113-9116
BACKGROUND: Arterial injury surrounding the mandibular angle is very common during mandibular angle ostectomy. The retromandibular vein is susceptible to injury owing to thin vascular wall and deep location. Anatomical studies regarding the retromandibular vein are rarely reported.OBJECTIVE: To observe the construction, branches, backstreaming of the retromandibular vein and its relation to the facial nerve and mandibular angle.METHODS: Twenty formalin-fixed adult cadavers (40 samples) were randomly selected. Bilateral retromandibular veins and their branches were cleaned. The construction, branches, backstreaming of the retromandibular vein and its relation to the facial nerve and mandibular angle were anatomically observed.RESULTS AND CONCLUSION: The retromandibular vein originated constantly and provided an average length of (4.51±2.01) cm, with an average outer diameter of (0.52±0.28) cm at the origination part. A total of 34 samples showed that the retromandibular vein was formed by the union of the superficial temporal and maxillary veins, and six samples showed that the retromandibular vein originated from the superficial temporal vein. The retromandibular vein divided into two types according to the backstreaming and course: one type consisted of the anterior and posterior branches: the anterior branch mostly drained into the internal jugular vein and minorly drained into the external jugular vein; the posterior branch was joined by the posterior auricular vein and became the external jugular vein. Another type of retromandibular vein without the anterior and posterior branches directly connected with the posterior auricular vein and entered the external jugular vein. The retromandibular vein had the following branches (appearance frequency): facial transverse vein (88%), mandibular articular vein (98%), parotid veins (95%),anterior auricular veins (58%), styloid process vein (43%). 88% of the retromandibular vein was located in the superior and inferior trunks of the facial nerve and in the medial branches. The retromandibular vein connected with facial nerve trunk or major branches in a decussated fashion. 75% of the arc-shaped structures formed by the retromandibular vein and its branches coursed close to the inferior and posterior border of the mandibular angle, and 25% of the arc-shaped structures were not close to the lowest point of the mandibular angle. The thin periosteum between the venous wall and cortical bone was just the position of ostectomy line of mandibular angle. The retromandibular vein was easily injured no matter in vein dissection or ostectomy.
4.Preparation of nano-liposome enveloping Flos Magnoliae volatile oil
Journal of Integrative Medicine 2007;5(3):314-7
OBJECTIVE: To discuss the influences of the concentrations of sodium cholate, cholesterol and drug, preparation temperature, ultrasonic time and power on the particle size and encapsulation efficiency of Flos Magnoliae volatile oil nano-liposome through film-ultrasonic wave technique together with membrane filter method. METHODS: The nano-liposomes were prepared by film-ultrasonic wave technique together with membrane filter method. The drug content in liposomes was determined by gas chromatography and the particle size was determined by laser light scattering granularity equipment. RESULTS: By film-ultrasonic wave technique together with membrane filter method, the liposomes in the range of 30-40 nm were prepared. The prepared nano-liposomes demonstrated good decentralization and homogeneity, and the encapsulation efficiency was (91.7+/-3)%. CONCLUSION: Film-ultrasonic wave together with membrane filter method is a simple method to fabricate nano-magnoliae liposome.
5.Clinical cases and prognosis analysis of primary gastrointestinal stromal tumors
Licheng LIU ; Xin WU ; Wentong XU
Chinese Journal of Digestion 2013;(3):150-154
Objective To analyze the clinical characters,therapeutic effect of gastrointestinal stromal tumors (GIST),the effects of surgical approach and imatinib adjuvant therapy on it.Methods From January 2004 to April 2010,the clinical data of 214 patients with primary GIST were retrospectively analyzed.The effects of surgical approach and imatinib on the survival of the GIST patients were compared.Count data were analyzed by chi-square test,and the survival rates were analyzed with the life table method and Kaplan-Meier curve.Results A total of 214 GIST patients'survival rates of 1-year,3-year,5-year were 93.0%,87.0% and 80.0%.According to the risk classification of National Institutes of Health,the differences of overall survival rates after surgery were statistically significant (x2 =22.058,P<0.05).The differences of survival rate among different nuclear division number were statistically significant (x2=26.599,P<0.05).The survival rate of pathological nuclear division number over 10/50 high power field (HPF) was the lowest.The overall survival rate of patients with gastrointestinal GIST was higher than patients with extragastrointestinal GIST,and the difference was statistically significant (x2 =68.139,P < 0.05).After complete resection of the tumor,the survival rate of patients with local recurrence was higher than those with widely recurrence,and the difference was statistically significant (x2 =4.409,P< 0.05).After complete resection of the tumor,the survival rate of patients taking imatinib was higher than that of patients not taking imatinib.Conclusions The surgical treatment was the main therapy for GIST.Complete resection of the tumor and taking imatinib after surgery could improve the prognosis and the survival rate of the GIST patients.
7.Expression and purification of Ebola virus trimeric glycoprotein based on novel glycoengineered Pichia pastoris
Musheng WU ; Xin GONG ; Shaohong CHANG ; Bo LIU ; Jun WU
Military Medical Sciences 2017;41(5):352-358
Objective To express and purify mammalian glycosylation modified trimeric Ebola virus trimeric glycoprotein (EBOV-GP) using novel glycoengineered Pichia pastoris.Methods The EBOV-GP and EBOV-GPΔMLDΔTM genes were cloned into the pPICZ-αA vector,electrochemically converted to glycoengineered Pichia pastoris,and compared with EBOV-GP expressed in HEK-293T cells.Glycosylation was analyzed by PNGaseF and EndoH digestion,while the target protein was purified by affinity chromatography and ion exchange chromatography.N-terminal sequencing was used to determine whether the signal peptide was correctly cleaved during protein translation and gel column analysis was used to find out whether the trimeric structure was formed.Results The results of PNGaseF showed that EBOV-GP expressed by glycoengineered Pichia pastoris and HEK-293T cells had the same relative molecular mass and N-glycosylation degree.EndoH digestion showed that the N-glycosylation modification of EBOV-GPΔMLDΔTM was in a non-high mannose form.N-terminal sequencing showed that the signal peptide of the GP protein itself was correctly excised.Gel column analysis showed that the purified protein was in a trimeric form.Conclusion An EBOV-GP is obtained with complex glycosylation modification based on Glycoengineered Pichia pastoris.
8.Intelligent Rotational Behavior Monitor for Experiment Animal
Xin LIU ; You WU ; Yan WANG ; Hong SHA
Chinese Journal of Comparative Medicine 2009;19(3):64-68
Objective to develop a rotation monitoring device to monitor animal rotation during experiments.Methods Use serial port controls,USB and rotary encoder et al.modern computer cience,with the combination of traditionally experimental methods and the software designed in VC language.Results A mini animal rotation monitor was developed,which could continuously monitor animal rotation and intelligently display the rotaion.conclusion Statistic analysis of experimental data proofs the rationality and effectiveness of the apparatus.The rotational behavior monitor can play an importance role in research into the pathogenesis of Parkinson's Disease(PD)and screening the anti-PD drugs.
9.Expressions of p75NTR, Bax and Bcl-2 and cell apoptosis in rat cortical neurons following mechanical injury
Haichun LIU ; Tao XIN ; Kaiyun YANG ; Wenliang WU ; Yunzhen CHEN
Chinese Journal of Trauma 2012;28(2):179-183
ObjectiveTo study the expressions of p75NTR,Bax and Bcl-2 and cell apoptosis in rat cortical neurons following mechanical injury and discuss the mechanism and mutual action way during the apoptosis of rat neurons after mechanical injury.Methods Cortical neuron cultures were prepared from the brain tissues of day 17 rat embryos and were exposed to mechanical injury seven days after seeding.After the traumatic neuron injury models were created,the apoptosis ratio of neurons was tested at several time points.The models were divided into the minor,moderate,severe injury groups according to the injury severity and the control group.The expressions of Bax and Bcl-2 in each group were detected by immunohistochemistry method and that of p75NTR by Western-blot.Combining with cellular apoptosis ratio in each group shown after FCM analysis,the correlation between the expressions of p75NTR,Bax and Bcl-2 and the apoptosis in rat neurons after mechanical injury could be analyzed.ResultsThe apoptosis ratio of the neurons in all the injury groups was obviously higher than that in the control group,with significantly higher apoptosis ratio of the neurons in the severe injury group than the minor and moderate injury groups (P <0.05).P75NTR,Bax and Bcl-2 were all expressed in all the injury groups,with statistical differences between groups.The expression of Bax in the severe injury group was significantly higher than that in the minor and moderate injury groups(P < 0.05 ). Conclusions p75NTR expression and Bax/Bcl-2 ratio are closely correlated with neuron apoptosis.The early expression of p75NTR may be one of mechnisms for neuronal apoptosis after neuron injury,when Bax and Bcl-2 may be involved.
10.A Survey of Medication Taken by Chinese Athletes
Jingzhu WANG ; Moutian WU ; Yinong ZHANG ; Xin LIU ; Zhiyong YANG
Chinese Journal of Sports Medicine 2004;23(4):447-450
Objective To make a survey of medication taken by Chinese athletes and a comparison between Chinese athletes and athletes from other countries in order to get information about how to improve Chinese athletes' performance. Method The information came from the forms"Doping Control Sample Collection" in which athletes answered the question: "What medications have you taken in the past 3 days?" The medicines taken by athletes were classified and statistically analyzed.Results 2,330 athletes and 25 kinds of sports were involved in. Medicines were statistically analyzed with 4 classes: profiling of declaration, vitamins and minerals, medicines for treatments, alternative medicine. Conclusion The survey recorded the types of supplements and medications taken by athletes in China in 1999. Chinese athletes took less vitamins and more alternative medicines than athletes from other countries.