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Objective To establish a quantitative polymerase chain reaction(PCR)method for the analysis of human-derived SRY DNA in mouse tissues,and to study the tissue distribution of human umbilical cord mesenchymal stem cells(HUCMSCs)in immunodeficient NOG mice after a single intravenous injection.Methods We established a quantitative PCR method for the analysis of human SRY DNA in mouse tissues,and validated the standard curve,linear range,accuracy,precision,and stability.Thirty-six NOG mice(18 male,18 female)were administered 3.5×107 HUCMSCs/kg by single intravenous injection.Six mice were then anesthetized and dissected after blood collection(EDTA anticoagulation)at 6,12,24,and 72 h,and at 1 and 2 weeks,respectively.DNA was extracted from lung,kidney,heart,liver,brain,spinal cord,stomach,small intestine,fat,skin,spleen,testis,uterus,and ovary tissues,and the distribution of HUCMSCs in each tissue was determined by the validated quantitative PCR method for detecting the human-derived SRY gene in mouse tissues.In addition,18 NOG mice(9 male,9 female)were divided into control(n = 6)and treatment groups(n = 12)injected intravenously with 0.9%sodium chloride and 3.5×107 cells/kg,respectively.Acute toxic reactions were observed during the administration period,and four animals were dissected at 72 h and at 2 and 4 weeks after administration to observe the gross organs.Mitochondrial protein expression was detected in paraffin sections of lung tissues by immunohistochemistry to analyze the colonization of HUCMSCs in lung tissues.Results The established RT-qPCR method for human-derived SRY DNA in mouse tissues met the validation criteria for each index.After a single intravenous injection in NOG mice,HUCMSCs were mainly distributed in the lungs and blood within 1 week after administration,with higher concentrations in lung tissues than in blood.The concentrations of HUCMSCs in lung tissue and blood remained relatively stable within 6~24 h and 6~72 h,respectively,and then decreased over time.The distribution of HUCMSCs in other tissues was not measured at all sampling points.The colonization result showed that HUCMSCs were detected in lungs 72 h after intravenous injection,but not at 2 and 4 weeks.No obvious acute toxicity was observed in NOG mice after single intravenous administration of HUCMSCs.Conclusions The above method for analyzing the distribution of HUCMSCs in mouse tissue is reliable and feasible.HUCMSCs were mainly distributed in lung and blood in NOG mice within 1 week after a single intravenous injection,and mainly colonized lung tissue at 72 h.A single intravenous administration of HUCMSCs has a good safety profile.
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Objective To detect the nucleation-promoting potency of retinol binding protein (RBP) in the simulated bile system,and observe the process of cholesterol nucleation and crystallization.Methods Small and synthetic bile systems were synthesized.RBP (RBP group),α1-acid glycoprotein (AAG,AAG group) and albumin ( Alb,Alb group) were added into the Small bile respectively.The derived indices of nucleation time,crystal growth rate and final concentration of crystal were studied by Busch method.Alb ( control group) and RBP (experimental group) were added into the Small and synthetic bile systems respectively.The nucleation time was detected by polarizing microscopy according to the Holan method and the nucleation-promoting potency was calculated according to the Holzbach method. The morphological dynamics of cholesterol nucleation and crystallization were observed under a light microscope.All data were analyzed by using the analysis of variance or the t test.Results The derived indices of nucleation time,crystal growth rate and final concentration of crystal were 0.66,1.29 and 1.01 in the AAG group,and 0.73,1.02,0.95 in the RBP group,respectively.In the Small bile system,the average nucleation time of cholesterol monohydrate crrystals were ( 12.2 ± 1.2 )days in the control group and (8.2 ± 1.5)days in the experimental group,respectively,with a significant difference between the 2 groups (t =2.97,P < 0.05 ).The nucleation activity of RBP was 0.67 in the Small bile system.In the synthetic bile system,the average nucleation time of cholesterol monohydrate crystals were (9.5 ± 1.1 ) days in the control group and (7.5 ± 1.1 )days in the experimental group,respectively,with a significant difference between the 2 groups ( t =2.35,P < 0.05 ).The nucleation activity of RBP was 0.79 in the synthetic bile system.The process of cholesterol nucleation and crystallization was divided into 4 stages,and the split phenomenon appeared in the bile system at day 21.Conclusion RBP is a pro-nucleation factor in simulated bile system.
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0.05).In the elderly group,every complication rates of LR were lower than those of OR(P
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Objective To observe the effects of endovascular radiation (ER) on the proliferation and apoptosis of medial smooth muscle cells (SMC) and to discuss the possible mechanisms of radiation in the prevention of vascular restenosis (RS) in rabbits after carotid endarterectomy (CEA).Methods Forty rabbits undergoing CEA were randomly divided into four groups (each group=10) and given a radiation dose of 0, 10, 20 and 40 Gy 32 P respectively. Rabbits were killed on the 3rd, 7th, 14th, 28th and 56th day after operation. The specimens were collected and histopathologic examinations were done.Results Proliferation apparently occurred in the intima and media of carotid the lumen became narrow in the control group on the 14 th, 28 th and 56 th day after operation. While in the radiation groups, proliferation was apparently suppressed and the lumen was much less narrowed ( P
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Objective To investigate the relation between CD44v6 and lymph node metastasis and prognosis of gastric carcinoma. Methods The expression of CD44v6 in 100 cases of advanced human gastric carcinoma was detected by immunohistochemical two-step method. Its relation with clinical pathology and prognosis was analyzed. Results CD44v6 expression occurred in 64%(64/100) of all gastric carcinomas; with the increase of depth of cancer invasion and local lymph node metastasis, the expression of CD44v6 elevated. Conclusion CD44v6 plays an important role in lymph node metastasis of human gastric carcinoma. It may be used as a new indicator to predict metastatic potential and prognosis of gastric carcinoma.
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Objective To study the correlation of the number and ratio of involved lymph nodes and the prognosis of gastric carcinoma .Methods 174 patients with positive gastric lymph nodes were included,they were treated by radical gastrectomy and lymph nodes dissection (D2 or D3)in our department. The number and ratio of metastastic gastric lymph nodes were calculated. Their influence and their interrelation with pathogenic parameters were analyzed. Results The overall 5 year survival rate was 29.59%, and the five year survival rates of patients with 1 20 per cent and more than 20% positive lymph nodes were 70.58%,11.86% respectively, one to five positive lymph nodes and five more were 46.47%,17.35% respectively, the five year survival rate was decreased as the ratio and number of involved lymph nodes increased. Studies also indicated that with the increase of depth of cancer invasion ,the number and ratio of involved lymph node elevated.Conclusions The results shows that the number and ratio of involved lymph nodes correlated to tumour progression and the five survival rate.Perigastric lymph node status can be considered as a simple and effective prognosis factor.
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Objective To summarize the experience of diagnosis and management of bleeding with GIVM.Method The clinical data of 26 patients admitted from 1989 to 1999 were analyzed retrospectively.Results GIVM with bleeding were revealed by endoscopies (42%),by ECT (55%),by selective arteriography (83%),by barium radiography (0%),and by exploratory laparotomy(100%).1 case was cured by conservative treatment.Bleeding intestinal segment was resected in 14 cases.4 cases were cured by interventional therapy.The others were cured by sclerosing agent injection,or by suture ligation of the dilated vessel or arterial gastroduodenalis.Conclusions Endoscopy is the first choice in the bleeding from GIVM.ECT may beused later if endoscopy is useless.Selective arteriography is the most valuable method in GIVM diagnosis.If necessary, exploratory laparotomy and intraoperative endoscopies may be used.Resecting bleeding segment is the most effective treatment.Other methods such as interventional therapy and suture ligation of dilated vessel are also effective.
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Objective The infiltration and micrometastasis of cancer cells via lymph vessel and blood is the important factors which influence the prognosis of patients with cancer. The present study was to establish the method of cytological examination of circulating gastric cancer cell in the peripheral blood and to explore its clinical significance. Methods The monocytes in blood samples, taken from 24 patients with advanced gastric cancer, were seperated by using Ficoll density gradient centrifugation and magnetic cell sorting system in which the magnetic microbeads were wrapped with cytokeratin 7 and 8. The samples were smeared and observed with HE stain. The expression of CD 34 , CD 45 , CEA and human telomerase reverse transcriptase (hTERT) of the cells were also examined with immunocytochemical and immunofluorescence staining methods. Results Gastric cancer cells were found in 10 out of 24 cases, and the positivity rate was 41.7%, which was significantly different from pathological differentiation of primary focus. Conclusion This method can be used to detect cancer cells from peripheral blood of patients with gastric cancer and therapeutic protocol and prediction of prognosis can be made based on it.
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Background and purpose:Spleen Tyrosine Kinase(Syk) may play a significant role in tumor signal transduction.Some studies showed that Syk was an anti-oncogene against oncogene Her-2.We investigated the impact of Syk expression on metastasis and apoptosis of gastric cancer cells in order to explore the new target therapy for the treatment of gastric carcinoma.Methods:Expression of Syk was evaluated by RT-PCR in 44 human gastric cancer tissues and adjacent normal gastric tissue samples,and apoptosis index in gastric carcinoma was detected by flow cytometry.The relationship between apoptosis index in gastric carcinoma and Syk expression was analyzed.Results:All normal gastric tissues demonstrated expression of the Syk gene,while 34 out of 44 gastric cancer tissues did not show any detectable Syk mRNA expression,there was a significant difference between the two groups(?2=9.14,P
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Objective:To determine the expression level of bone morphogenetic proteins-4 (BMP-4) and the relationship between the expression of BMP-4 and the clinical/pathological parameters in patients with gastric cancer. Methods:Using im-munohistochemical staining technique ,expressions of BMP-4 were investigated in different tissues from 90 gastric cancer specimens and 30 specimens from normal gastric mucosa as control. ResultS:The positive rate of BMP-4 was 23. 3% (21/90) in gastric cancer and 3. 3% (1/30) in normal gastric mucosa. The expression of BMP-4 in gastric cancer was closely related with locus, serosa infiltration,cell differentiation and lymph node metastasis (P
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Objective To find a novel operative modality with sphincter preservation in the treatment of middle and low rectal carcinoma. Methods Pull through lower resection was performed on 28 rectal cancer patients. The distance between the anal verge and the lower margin of the tumor was 6~8cm(20 patients) or 8~10cm(8 patients), including 8 patients in Dukes A stage, 16 Dukes B and 4 Dukes C. The resected line from tumor distal margin was 2cm, 3cm, and 4cm, respectively. Results There was no operative death, anastomotic fistula or anastomotic stenosis in these cases. Mean follow up period was 30 months. Local recurrence was found in two cases (7.1%) 18 months after the operation, and 26 cases were cancer free till the end of the follow up. Defecation was satisfactorily controlled 8~12 weeks after the operation. Conclusions Pull through Welch procedure could meet the criterion of the radical resection of rectal carcinoma,and keep the internal and external sphincter muscles intact in the superior lower anterior resection. The normal defecationcan can maintain after the operation due to the preservation of internal and external sphincter muscles.
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Objective To evaluate carotid endarterectomy for patients with atherosclerotic occlusive diseases. Method Surgical experience of endarterectomy in 32 patients with atherosclerotic occlusive diseases was retrospectively analyzed. Internal carotid artery stem pressure was measured during operation. Shunting was used routinely. Patching was used electively. Anti-platelet agents and agents reducing plasma fibrinogen were given perioperatively. Result There was no perioperative mortality, nor ischemic cerebral stroke. Patients were followed-up from 5 months to 20 months with no occurrence of ischemic cerebral stroke during the follow-up. One case presented dysfunction of hypoglossal nerve. Two cases presented dilation of carotid artery on ultrasound scan. There were no restenosis (≥50%) and thrombosis. Conclusion Carotid endarterectomy for patients with atherosclerotic occlusive diseases of carotid artery is effective and safe. Careful performance, shunting, patching and the correct order of clamps removing are the keys of preventing complications.
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Objective To explore the diagnosis and management of ruptured abdominal aortic aneurysm(RAAA).Methods Twelve patients with RAAA treated in past 7 years were revienled retrospectively.The main clinical manifestations were abdominal pain and / or back pain,low blood pressure or shock,and pulsating abdominal mass.All cases were accurately diagnosed with CT and 7 were treated by conventional operation,one by EVAR,and the other 4 did not receive surgical treatment.Results Perioperative death occurred in 5 cases(mortality rate was 62.5%) in 8 surgical treated patients,including circulatory failure in 2 cases,renal failure in 1 case,and multiple organ failure in 2 cases.All the 4 patients treated with nonoperative method were dead.Conclusions Surgical operation in RAAA cases still carried a high mortality.Early dignosis,appropriate resuscitation,urgent surgical repair,reduction of operative time,and infrarenal clamping are measures conducive to lowering the mortality rate of RAAA.EVAR has the potential to reduce the mortality rate from RAAA.
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Objective:To find the possible mechanisms of endovascular radiation in the prevention of restenosis in rabbits following carotid endarterectomy and to observe the dose effect relationship.Methods:Twenty four rabbits were randomly divided into three groups following carotid endarterectomy (each n=8) and were allocated to receive a radiation dose of 0,10,20Gy( 32 P)respectively.Changes in the levels of plasma nitric oxide(NO) and endothelin 1(ET 1)were measured 3 day before operation and 3,7,14,28 day after operation.Results:The plasma NO was markedly increased and ET 1 was markedly decreased in endovascular radiation groups compared to control groups.The results were similar in the two groups(10Gy versus 20Gy).Conclusion:The endovascular radiation may inhibit smooth muscle cell proliferation and migration,and thus prevent restenosis by increasing NO and decreasing the level of ET 1.Changes of the levels of plasma NO and ET 1 is a practicable methods to detect the effect of therapy.