Gene transfection is an important method for studying gene function. The origination and development of gene transfection are reviewed in this article. Six methods of gene transfection are introduced, among which, special attention are given to two new ones: particle bombardment and photochemistry transfection technology. Besides, the advantages and disadvantages of these methods are compared in order that more suitable methods can be chosen in researches. The application of the technology in gene research is discussed in the end of this article.

Objective To investigate the relationshp among Gp Ⅱ b/ⅡⅢ a、CD62p in platelet activation mark-er and high-sensitivity C-reactive protein (hs-CRP) in elderly patients with chronic cor pulmonary exacerbation. Methods The subjects were divided into four groups (42 elderly patients with cor pulmonary exacerbation,42 eld-erly patients with cot pulmonary remission,30 cases of healthy elderly subjects and 30 cases of healthy non-elderly subjects) . The expression of Gp Ⅱ b/Ⅲ a and CD62p were measured with tricolor flow cytometry. Hs-CRP levels were measured by nephelometry and turbidimetry in patients and normal controls. Results Compared with cor pul-monary remission group,healthy elderly group and healthy non-elderly group, the levels of Gp Ⅱ b/Ⅲ a, CD62p and hs-CRP increased significantly in patients with chronic cor pulmonary exacerbation ( all P < 0.001 ). Gp Ⅱ b/Ⅲ a and CD62p results of the elderly patients with cot pulmonary remission were higher than those in healthy elderly subjects (P <0.05) and healthy non-elderly subjects (P < 0. 001 ) . The expressions of both Gp Ⅱ b/Ⅲa and CD62p in healthy elderly subjects were statistically higher than that in healthy non-elderly subjects ( P < 0.05 ). There was a positive con'elation between the Gp Ⅱ b/Ⅲ a、CD62p and hs-CRP in elderly patients with chronic cot pulmonary exac-erbation(r =0.59,P<0.01 ; r=0.54,P<0.01 ). Conclusions The platelet in elderly patients with chronic cot pulmonary exacerbation is overactivated,which is correlated with hs-CRP.

Objective　To evaluate the treatment of in-stent restenosis with rotational atherectomy and balloon angioplasty. Methods　The rotational atherectomy and 4～6 atm low pressure balloon angioplasty was performed in 3 patients with in-stent restenosis and follow up after treatment. Results　All cases were succeeded. The bradycardia occurred in one patient was quickly disappeared without treatment, two other patients were found no effect on heart rate, hemodynamic performance, global LV function, or regional wall motion. No complications, angina, death or other coronary event occurred during the follow up for 6～12 months. Two of them was performed coronary angiography after 6 months and showed the diameter of target vessel was less than 30% as compared with that on coronary angiography which performed immedately after operation. Conclusion　The management of in-stent restenosis in target vessels using a combination of rotational atherectomy and balloon angioplasty is safe and efficient.

Objective This study was designed to evaluate the effect of laparoscopic-assisted subtotal colectomy(LASC)and modified Duhamel procedure in the treatment of severe functional constipation(SFC).Methods From June 2005 to October 2007,100 cases of SFC were treated by LASC combined with modified Duhamel procedure in our institute.After freed the colon under a laparoscope,we made a small incision in the lower abdomen to perform subtotal colectomy while preserved the ileocecal region.Then a side-to-side colorectal anastomosis was conducted.Results In this study,98 cases of LASC were successively completed and 2 cases were converted to open surgery.The operation time of LASC in the 98 cases ranged from 48 to 125 min mean,(76?23)min],and the total operation time was 140-225 min mean,(170?42)min].The intraoperative blood loss was 10-160 m1 mean,(35?18)m1].In the patients,the intestinal function was recovered within 25-52 hours mean,(42?12)h] after the operation;none of them developed postoperative wound infection,anastomotic stoma,or other complications.One week after the operation,2 of the 100 patients showed mild dehydration,and 2 were diagnosed as having incomplete small bowl obstruction.All of the four were cured by conservative treatments.The other 96 patients who had no complication were discharged from the hospital in 5-11 days mean,(7.0?1.5)days] postperation.One year later,the symptoms of constipation were significantly alleviated(defecation 1-3 times/d)in 90 patients;7 cases had various degrees of diarrhea(defecation 4-6 times/d);and 3 patients showed recurrence of mild constipation(defecation 1 time/2-3 d).The gastrointestinal quality of life index(GIQOL)determined in 1 year postperation was(109?20)points,which indicated good results compared to(94?23)points before the operation(maximum:144).Eighty-three cases were satisfied with their frequency of defecation.Conclusions LASC combined with modified Duhamel procedure is an effective method for the treatment of SFC,resulting in less damage,less influence to the internal environment,and quick recovery.The procedure is worth being wildly used.

Objective\ To introduce the speciality of pontine infaction in clinic and screenage.Method\ we made retrospective study on clinical data and image data of pontine infarction caused by BAD,and compared with the data of 31 patients with lacunar infarction in pontine.Result\ Clinical spetiality of BAD group:There are a few disturbance of consciousness,mainly the movement disturbance and dysarthria with ocular movement disturbance.Compared with the control group,there is difference between the two groups(P

Objective To evaluate the clinical efficacy of dexmedetomidine given intranasally for day surgery recovery period.Methods 60 pediatric patients who were treated for elective day surgery were selected as study objects,and they were randomly divided into control group and observation group according to the digital table,30 cases in each group.During operation, after induction of 8% sevoflurane inhalation at 8 L/min,3% of sevoflurane was maintained at 1 L/min.After intubation,0.5 g dexmedetomidine was given intranasally in the observation group, 0.9% NaCl saline was given to the control group.Mean arterial pressure (MAP),heart rate (HR),oxygen saturation (SpO2) were observed at different time points.The recovery time, number of cases of agitation, Aono agitation scale, children anesthesia delirium PAED score and the incidence of adverse reactions were observed in the two groups.Results As the time passed by, the MAP,HR,SpO2 in the two groups stabilized, except HR of the observation group at T0 time was higher than the control group[(92.92±3.22)beats/min vs.(89.32±3.22) beats/min], the difference was statistically significant(t=2.34,P<0.05).The children of the observation group recovered more slowly than the control group[(20.55±0.97)min vs.(18.62±1.03)min,t=2.17,P<0.05)],the number of cases of agitation, Aono restless score, anesthesia delirium PAED scores were lower than children in the control group (t=5.38,3.72,2.56,all P>0.05).The overall incidence of adverse reactions was similar in the two groups, there was no statistically significant difference (13.33% vs.10.00%,x2=0.12,P>0.05).Conclusion Dexmedetomidine given intranasally for the children underwent day surgery can significantly improve the quality of children awakeness, reduce the incidence of postoperative delirium, and without significant side effects.

Objective To understand the effect of astragalus on serum levels of advanced glycation end products (AGEs),soluble receptor for advanced glycation end products (sRAGE) in newly diagnosed patients with type 2 diabetes.Methods 60 patients of newly diagnosed type 2 diabetes were randomly divided into control group (28 patients) and treatment group (28 patients),the control group was given control of blood glucose,blood pressure,lipids and other necessary treatment,and the treatment group was additionally treated with astmgalus granule based on the control group.The course for both treatments is 12 weeks.Various clinical and biochemical parameters were tested before and after treatment in both groups with enzyme-linked immunosorbent assay for the detection of serum AGEs,sRAGE levels.Results ① Glycosylated hemoglobin (HbA1c) in the control group after treatment (7.28± 0.83)％ was significantly decreased than before treatment (9.57±1.14) ％,the difference was statistically significant (t＝5.75,P＜0.05); glycerin three greases (TG) in the control group after the treatment (1.45± 0.39)mmol/L decreased significantly compared with those before treatment (1.92± 1.01)mmol/L,the difference was statistically significant(t＝2.79,P＜0.05); ② HbA1c (7.16± 0.88) ％ in the treatment group after treatment was significantly decreased than before treatment (9.29± 1.62)％,the difference was statistically significant (t＝6.08,P＜0.05) ; TG (1.53 ± 0.41) mmol/L in the treatment group after treatment was significantly decreased compared with those before treatment (2.11 ± 0.79)mmol/L,the difference was statistically significant (t ＝ 2.40,P ＜ 0.05) ; the levels of serum sRAGE (20.38 ± 8.01) ng/ml in the treatment group after treatment increased obviously compared with those before treatment (15.10 ± 9.22)ng/ml,the difference was statistically significant (t＝-2.29,P＜0.05); ③ The level of serum sRAGE (20.38 ± 8.01) ng/ml in the treatment group after treatment increased obviously than the levels of serum sRAGE (15.13 ± 9.27)ng/ml in the control group after treatment,the difference was statistically significant (t＝-1.17,P＜0.05).Conclusion Astragali can increase serum sRAGE expression in newly diagnosed type 2 diabetic patients.

Objective To understand the serum levels of advanced glycation end products (AGEs),soluble receptor for advanced glycation end products (sRAGE) in newly diagnosed patients with type 2 diabetes.Methods 60 patients of newly diagnosed type 2 diabetes were chosen from 2010 September to 2011 December in Department of Endocrinology Changshou Chinese medicine hospital in Jiangsu Province,and 30 healthy people were screened as control group.Various clinical and biochemical parameters were detected,using enzyme-linked immunosorbent assay for the detection of serum AGEs and sRAGE levels.Results The levels of serum AGEs in newly diagnosed type 2 diabetic patients and control group were (1379.2 ± 431.8) and (1154.5 ±326.4) pg/ml,and there were significant differences between two groups (P＜0.05).The levels of serum sRAGE in newly diagnosed type 2 diabetic patients and control group were (14.6± 9.3)and (19.5 ± 8.9)ng/ml,and there were also significant differences between two groups (P＜0.05).The level of serum AGEs in newly diagnosed type 2 diabetic patients was positive correlation with glycosylated hemoglobin (HbA1c),and the level of serum sRAGE was associated with low density lipoprotein cholesterol (LDL-C) negative correlation.Conclusion The level of serum AGEs increase in newly diagnosed type 2 diabetic patients,and the level of serum sRAGE decrease in newly diagnosed type 2 diabetic patients.

Objective To determine the inhibitory effect of a synthetic hexa-peptide GGWSHW (G peptide) derived from thrombospondin-1 (TSP1) on TGF-? activation induced by angiotensin Ⅱ (Ang Ⅱ) in cultured human renal tubular epithelial cells. Methods Human proximal tubular epithelial cell line (HK-2) was cultured in vitro, untreated HK-2 cells were acted as normal control group. HK-2 cells were then stimulated by Ang Ⅱ for 1-24 hours (Ang Ⅱ stimulation group),so that optimal dosage and duration could be chosen.One hour prior to induction,HK-2 cells were pretreated with 10 ?mol/L peptide G (G peptide treated group)or losartan (losartan treated group), the blocker of I type receptor of Ang Ⅱ was acted as inhibitory control.The mRNA and protein levels of TSP1,TGF-?1,FN and PAI-1 were measured by RT-PCR and Western blot. Confocal microscopy and flow cytometry were performed to detect the presence of TSP1, TGF-?1 and co-positive expression of two protein, respectively.The concentrations of total and active TGF-?1 as well as FN and PAI-1 in cell culture supematants were measured by ELISA. Additionally, the expression of Smad2 and p-Smad2 was also examined for the bioactivity of TGF-?1 signaling protein. Results Ang Ⅱ enhanced the expression of TSP1 and TGF-?1 in a temporal-spatial dependent manner. The optimal dosage and duration were 1 ?mol/L and 6 hours,for TSP1,and 0.1 ?mol/L and 12 hours for TGF-?1 respectively.Comparing with untreated HK-2,the co-expression of TSP1 and TGF-?1 induced by A Ⅱ showed a increase of 5.4 folds.In addition,the protein level of p-Smad2 was elevated remarkedly, the mRNA level of FN and PAI-1 was up-regulated by 3 and 1.5 folds, and the concentration was increased by 2.0 and 1.9 folds respectively. Peptide G had less effect on the expression of TSP1 and TGF-?1,whereas it significantly reduced the secretion of active TGF-?1,though total level of TGF-?1 remained up-regulated. Furthermore,comparing with losartan treated group, p-Smad2 expression was reduced by 28.9%, the mRNA level of FN and PAI-1 was decreased by 34.5% and 26% respectively,and the protein levels were reduced by 11.0% and 8.9% respectively. Conclusion The inhibitory peptide derived from TSP1 effectively suppresses TGF-?1 activation through a competitive mechanism and also reduces the secretion of FN and PAI-1 associated with fibrosis.

Objective To understand the effect of astragalus on insulin resistance and plasma amylin levels in newly diagnosed type 2 diabetic patients.Methods 88 patients of newly diagnosed type 2 diabetes were randomly divided into three groups:Group A (lifestyle intervention group) contained 30 patients,Group B (metformin treatment group) also contained 30 Patients,and Group C (astragalus and metformin treatment group) contained 28 Patients.Patients in group A were intervened with the control of diet,blood pressure and lipids level; patients in group B were additionally treated with metformin on the basis of group A; patients in group C were additionally treated with metformin and astragalus on the basis of group A.The course for both treatments were 8 weeks.Various clinical and biochemical parameters were detected before and after the treatment in all three groups and enzyme-linked immunosorbent assay was adopted for the detection of plasma amylin.Results Fasting blood glucose levels were significantly decreased after treataent in the three groups (t=-2.696、-4.029、-3.995,P＜0.05) ; insulin resistance index reduced in group B and group C (t=-2.599、-3.813,P＜0.05),the difference between group C and group B was statistically significant (t=-2.334,P＜0.05) ;treatments of group B and group C could improve the beta cell function index (t=2.303、2.384,P＜0.05),and they also could increase the plasma amylin level (t=2.341、3.045,P＜0.05).Conclusion Astragalus and metformin can improve insulin resistance index and increase plasma amylin levels in newly diagnosed type 2 diabetic patients.