Effects of cisplatin-alginate conjugate liposomes in cervical cancer-bearing nude mice / 上海交通大学学报（医学版）

Objective · To establish human cervical cancer xenografts in nude mice and investigate the antitumor therapeutic effects and safety of EGF modified cisplatin-alginate conjugate liposomes. Methods · Cervical cancer-bearing mouse model was established by subcutaneously injecting Hela cells in nude mice. Generalstate and xenograft growth of the mice were observed. Tumor volumes, tumor weights, and the body weights of mice during the treatment were recorded and analyzed. The expression levels of EGFR in xenografts were detected by immunohistochemistry. Results · ① EGFR was highly expressed in the xenografts. ② CS-EGF-Lip inhibited the tumor growth effectively (P=0.000). ③ The inhibition rates of tumor volume and tumor weight of CS-EGF-Lip were 80.22% and 58.60% respectively, which were betterthan those of cisplatin (P=0.000). ④ CS-EGF-Lip had minimal influence on body weight in mice (P=0.000). Conclusion · CS-EGF-Lip has more effective antitumoreffects than cisplatin in cervical cancer-bearing mice, which can inhibit tumor growth of solid tumors with enhanced efficacy and safety.

Effect of early-stage keep warm intervention on improving low temperature risk in children underwent blood purification treatment / 中国实用护理杂志

Objective To evaluate the clinical effectiveness of early-stage keep warm intervention on improving low temperature risk children treated with blood purification (BP). Methods Ninety children were randomized into observation group (46 cases) and control group of (44 cases) from July 2013 to September 2015. Control group were nursed with conventional BP standard operation process, while the observation group were nursed additionally with heat insulation blanket before 30 min of booting machine, and recorded the central body temperature of 0 min,30 min,60 min,90 min,120 min for each 60 min 1 time in the future. Until the end of the blood purification 60 min. Low temperature complications were recorded and judged between the two groups. Results In the observation group, 107 cases of low body temperature occurred during the course of 7 cases of blood purification, occurrence rate of 6.5% (7/107). The control group was 15.8% (16/101). The difference between the 2 groups was statistically significant (χ2=4.569,P<0.05);In the continuous blood purification group, observation group 10.6% (5/47), The incidence rate of control group was 29.3% (12/41), he difference between the 2 groups was statistically significant (χ2=4.876,P<0.05). Conclusions Application of heat insulation blanket at the early-stage may effectively reduce the risk of hypothermia complications in the children treated with blood purification. The continuous blood purification effect is more significant.

Protective effect of glucagon-like peptide-1 analogue on cardiomyocytes injury induced by hypoxia/reoxygenation / 中华内科杂志

Objective To investigate the effect of glucagon-like peptide-1 (GLP-1) receptor agonist liraglutide on hypoxia/reoxygenation (H/R)-induced cardiomyocytes death under high glucose condition and the potential mechanisms.Methods H9C2 cardiomyocytes were divided into 4 groups:normal glucose (N,5 mmol/L),high glucose (G,20 mmol/L),high glucose in combination with liraglutide (L,100 nmol/L),high glucose in combination with liraglutide and wortmannin (W,25 nmol/L).The apoptosis of H9C2 was detected by TUNEL assay.Nitric oxide synthetase(eNOS),nitric oxide (NO) and reactive oxygen(ROS) in supernatants were measured by enzymatic analysis,p-PI3K,PI3K,p-Akt,Akt,Bcl-2,caspase-3 were examined by western blotting.Results Compared with cells in N group,the apoptosis of H9C2 cells induced by H/R was markedly increased [(15.79 ± 3.92) ％ vs (9.74 ± 1.14) ％,P =0.028] in G group.The same was true for ROS [(489.63 ±21.01) U/ml vs (338.50 ±43.60) U/ml,P ＜0.001] and caspase-3 levels (1.87 ±0.03 vs 1.15 ±0.04,P ＜0.001),but not for Bcl-2 protein expression (1.79 ± 0.06 vs 1.89 ±0.03,P =0.047).Pretreatment of cells with liraglutide (100 nmol/L) prevented the cell death induced by high glucose and H/R together with decrease of ROS and caspase-3 levels and increase of Bcl-1 expression.Moreover,treatment of cells with liraglutide also significantly increased phosphorylation of PI3K and Akt (p-PI3K/PI3K:0.87 ± 0.07 vs 0.59 ± 0.09,P =0.002;p-Akt/Akt:0.34 ± 0.01 vs 0.08 ± 0.01,P＜0.001),eNOS[(41.29 ±0.56) μmpl/L vs (37.20 ±0.52)μxmpl/L,P ＜0.001]and NO [(31.24 ±0.40) μmpl/L vs (26.66 ±0.53) μmpl/L,P ＜0.001] levels.Furthermore,addition of PI3K/Akt inhibitor wortmanin markedly inhibited the expression of p-PI3K/PI3K,p-Akt/Akt,reversed the changes of eNOS,NO,caspase-3 and Bcl-2 by liraglutide,and abolished the protective effect of liraglutide on cell apoptosis.Conclusions GLP-1 receptor agonist liraglutide treatment could alleviate cardiomyoeytes apoptosis induced by high glucose and H/R through the activation of PI3K-Akt-eNOS-NO signaling pathway and inhibition of oxidative stress.

Effect of EMG-Triggered Stimulation on Drop Foot after Hemiplegia / 中国康复理论与实践

@#Objective To investigate the effect of EMG-triggered stimulation on drop foot for poststroke hemiplegia. Methods 36 hemiplegic patients were randomized into routine group (n=18) and EMG-triggered stimulation group (triggered group, n=18). 2 groups were assessed by Fugl-Meyer Assessment (FMA), range of motion (ROM) of ankle dorsiflexion and iEMG values before and after treatment. Results The scores of FMA, degrees of ROM, iEMG values of the tibialis anterior, and co-contraction ratio improved in 2 groups after treatment (P<0.05), and the triggered group had better effect than the routine group (P<0.05). Conclusion EMG-triggered stimulation can improve muscle strength, and decrease co-contraction ratio.

Reasons for accidental extubation on infants mechanical ventilation and optimization research of intervention countermeasures / 中国实用护理杂志

Objective To analyze the reasons for accidental extubation in infants during mechanical ventilation and to explore effective intervention countermeasures.Methods A total of 148 infants with accidental extubation from May 2013 to February 2014 who given routine nursing care were selected as control group.Another 152 infants with accidental extubation in pediatric intensive care unit (PICU) who given optimization nursing care from March to December 2014 were selected as observation group.Retrospective analysis of accidental extubation,artificial airway way and intervention care were conducted.Results The total intratracheal tube time was 283 d in control group and 253 d in observation group.Nineteen cases (6.7％) suffered accidental exudation in control group and that of 8 cases(3.2％) in observation group,and there was significant difference between two groups,x2=5.25,P＜0.05.The primary reason of unplanned extubation was that infants with unconsciousness,the second cause was the improper catheter fixed.The third cause was the patient's comfort level and so on the improper nursing operation leading to accidental extubation too short or too shallow.Conclusion Infants given mechanical ventilation should pay more attention to the intervention of accidental extubation,appropriate calm,physical constraints and correct fixed pipeline.

Effect of EMG-Triggered Stimulation on Drop Foot after Hemiplegia / 中国康复理论与实践

Objective To investigate the effect of EMG-triggered stimulation on drop foot for poststroke hemiplegia. Methods 36 hemi-plegic patients were randomized into routine group (n=18) and EMG-triggered stimulation group (triggered group, n=18). 2 groups were as-sessed by Fugl-Meyer Assessment (FMA), range of motion (ROM) of ankle dorsiflexion and iEMG values before and after treatment. Re-sults The scores of FMA, degrees of ROM, iEMG values of the tibialis anterior, and co-contraction ratio improved in 2 groups after treat-ment (P<0.05), and the triggered group had better effect than the routine group (P<0.05). Conclusion EMG-triggered stimulation can im-prove muscle strength, and decrease co-contraction ratio.

Genetic analysis of a congenital nephrogenic diabetes insipidus pedigree / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>As an X-linked recessive way, arginine vasopressin receptor 2 (AVPR2) gene mutation resulted in a hereditary disease - congenital nephrogenic diabetes insipidus (CNDI). We found a suspect clinical CNDI pedigree. In order to identify the genetic etiology, we performed the genetic analysis.</p><p><b>METHODS</b>The clinical features of the proband and his family members were recorded. The laboratory tests and imaging inspections were analyzed. The water deprivation and pituitrin loading test were performed in the proband and his brother. The genomic DNA of all the members of the pedigree was extracted and then PCR amplification on AVPR2 gene was carried out. Sequencing in both directions was performed to identify mutation on AVPR2 gene.</p><p><b>RESULTS</b>Both the proband and his brother were diagnosed as CNDI, meanwhile the other members of this pedigree were normal. No severe biochemical abnormality was found in the two CNDI patients. Both the patients had moderate urinary retention, severe megaloureter and hydronephrosis, and mild renal insufficiency. Two mutations of AVPR2 gene were discovered in the 3rd exon in the patients, a silent mutation L309L and a nonsense mutation R337X. The AVPR2 gene R337X mutation was co-segregated with CNDI. R337X mutation was not a reported mutation in the mainland of China.</p><p><b>CONCLUSION</b>The AVPR2 gene R337X mutation was also a genetic etiology of CNDI patients in the mainland of China.</p>

Preliminary Study on Traditional Chinese Medicine Treatment Characteristics of Intestinal Cancer based on Scale-free Network Analysis Method / 世界科学技术-中医药现代化

This article was aimed to study the characteristics of intestinal cancer by traditional Chinese medicine (TCM) based on the scale-free network analysis method. History information of 145 hospitalized cases from 2008 to 2011 was collected from the Oncology Department, Guang A nmen Hospital, in the clinical collection system. The rule of medication in the treatment of intestinal cancer by TCM was explored by scale-free network analysis method from several aspects, such as effects, classification and compatibility relations. The analysis results of 145 intestinal cancer cases showed that strengthening the body resistance was the main treatment principle. Si-Jun-Zi (SJZ) decoc-tion was used with the highest frequency. In the prescription design, qi-supplementing herbs were the most. The heat-clearing, dampness-eliminating, qi-regulating, blood-activating and stasis-removing, food stagnation removing and phlegm-removing herbs were also used frequently. The TCM treatment and syndrome differentiation will be changed due to the treatment and disease stages. It was concluded that scale-free network analysis method is able to relatively show the TCM treatment rule of intestinal cancer quantificationally and intuitively.

Study on Traditional Chinese Medicine Medication Rules in the Treatment of Gastric Cancer based on Scale-free Network Analysis / 世界科学技术-中医药现代化

This article was aimed to study medication rules of traditional Chinese medicine (TCM) in the treatment of gastric cancer. The history information of 111 hospitalized cases from 2008 to 2011 was collected from the Department of Oncology of Guang'anmen Hospital with the clinical collection system. The medication rules in the treatment of gastric cancer by TCM were explored by the scale-free network analysis from several aspects, such as effects, classifications and compatibility relations of Chinese medicine. The analysis result of 111 gastric cancer cases showed that strengthening the body resistance was the main treatment principle. Liujunzi decoction was the most frequently used prescription. In TCM compatibility, deficiency-nourishing herbs were the main ingredients, especiallyqi-supplementing herbs which take the first place in the prescription. And the heat-reducing herbs, dampness-eliminating herbs, blood-activating and stasis-eliminating herbs, food stagnation resolving herbs and qi regulation herbs were also used in the combination. Core herbs in the prescription for the treatment of gastric cancer were Tuckahoe, Codonopsis pilosula, Astragalus root, Pericarpium Citri Reticulatae, Fructus Aurantii, Nidus Vespae and etc. TCM treatment, syndrome differentiation and medication are varied according to different therapeutic stages and disease stages. The result showed that scale-free network analysis had certain significance in the understanding of medication rules of TCM treatment of gastric cancer.

Risk of cervical cancer and precancerous diseases in the oral HPV carriers / 中华妇产科杂志

Objective To evaluate the risk of the occurrence of cervical intraepithelial neoplasia (CIN) and invasive cervical cancer in the oral HPV carriers through a population-based investigation in Shanghai.Methods A total of 1200 cases of outpatients who attended the annual cervical examination and 50 preoperational cases of inpatients with CIN Ⅲ or invasive cervical cancer were enrolled from three clinical centers in Shanghai.The oral HPV infection was determined by real-time PCR.In 1200-case cross-sectional study,the incidence rate of CIN was compared between the oral HPV positive and negative cohort.In 1250-case case-control study,the positive rate of oral HPV DNA test was compared among normal control group,CIN Ⅰ-Ⅲ,and invasive cancer case groups,and all odds ratio (OR) values were calculated,respectively.The HPV transmission-related demographic and behavioral characters of the oral HPV carriers were also analyzed.Results The oral HPV carriers accounted for 5.9％ (71/1200) of the investigated outpatients.The oral HPV DNA positive rates were gradually increased with the cervical disease grades,which were 5.8％ (68/1182,normal),2/13 (CIN Ⅰ),1/5 (CIN Ⅱ),31.4％ (11/35,CIN Ⅲ) and 5/15 (invasive cancer).In cross-sectional cohort studies,the relative risks (RR) of CIN Ⅰ,Ⅱ were 2.9 and 4.0 for oral HPV carriers,respectively.In case-control study,the OR values for CIN Ⅰ-Ⅲ and invasive cervical cancer were 3.1(95％CI:1.6-10.1),4.2(95％CI:1.7-28.4),7.1(95％CI:4.8-19.8) and 10.1 (95％ CI:3.2-32.1),respectively.The oral sex and multi-sexual partner were two major risk factors for the oral and cervical HPV co-infection,HPV-related cervical cancer and precancerous diseases according to behavioral analysis.Conclusions There are complicated transmission pathways between oral and cervicalHPV.Oral HPV carriers should be intensively followed up and their oral HPV infection and HPV-related cervical diseases should be treated together.

The diagnostic value of one-stop liver magnetic resonance dynamic enhanced MRA for esophageal and gastric varces / 中国基层医药

Objective To investigate the diagnostic value of one-stop liver magnetic resonance dynamic enhanced MRA for esophageal and gastric varces.Methods 82 patients with liver cirrhosis portal hypertension who were diagnosed by clinical and laboratory detect were taken,conducting liver horizontal scanning and multitemporal three dimensional dynamic enhanced scanning using Philips Achieva 1.5T MRI scanner,and then make maximum intensity projection(MIP) at work station to draw up portal vein image,observing the form and distribution of portal vein and its branches,meanwhile refer to cross sectional image to carefully observe whether there was cirsoid vein in esophagus and stomach,and further the source,degree and depth of cirsoid vein were compared,and whether there was cirsoid vein in other parts,and the pathological situation of liver.Compared with the result of gastroscope inspection,whether there is cirsoid vein in esophagus and stomach.Results The trunk and class-Ⅲ branches of portal vein of all patients had been clearly shown.In this team,there were esophageal and gastric varces in 78 patients,specifically including varices of fundus of stomach & esophageal varices in 27 patients,and varices of fundus of stomach and varices of body of stomach in 19 patients.Meanwhile,it diagnose 6 patients with liver cirrhosis and liver cancer and indicate 8 patients with liver cirrhosis nodule cancerization.The endoscope found out esophageal varices and varices of fundus of stomach in 68 patients,specifically including gastric varicesa and esophageal varices in 25 patients,sole gastric varicesa in 8 patients,and sole esophageal varices in 35 patients.Conclusion The one-stop liver three dimensional dynamic enhanced MRA is with great value in diagnosing the location and degree of esophageal varices and varices of fundus of stomach of patients with portal hypertension.The overall effect is better than endoscope.

The effect of parathyroid hormone on expression of matrix GLA protein in ovariectomized SD rats and primary osteoblasts / 中华内分泌代谢杂志

ObjectiveTo observe the effect of parathyroid hormone on expression of matrix GLA protein (MGP) in ovariectomized SD rats and primary osteoblast,and to study the role of MGP on the possible mechanism of postmenopausal osteoporosis.MethodsThirty-six Sprague-Dawley female rats were allocated into 3 groups,12 in each:sham operation group,ovariectomized group( OVX group),ovariectomized and parathyroid hormone treatment group.Animals in the parathyroid hormone group were injected parathyroid hormone (20 μg/kg,three times a week for 12 weeks) three weeks after ovariectomy.All rats were sacrificed after 18 weeks.Urine and serum were collected every three weeks.Lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density (BMD) of lumbar vertebra of rats was determined.The content of MGP in serum and urine was determined by enzyme-linked immunosorbent assay (ELISA).Expression of undercarboxylated Matrix GLA Protein (ucMGP) was detected by immunochistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerause chain reaction.Results ( 1 ) 18 weeks after ovariectomy,BMD of lumbar vertebra in OVX group was lower than those in sham group and parathyroid hormone group significantly ( P＜0.05 ).(2) The content of MGP in serum and urine was dynamic variation after treatment hy parathyroid hormone,and it was significant compared with OVX group ( P＜0.05 ).( 3 ) Immunohistochemical localization of ucMGP was seen in lumbar vertebra in OVX group.(4) Relative quantification of MGP mRNA expression in lumbar vertebra in OVX group was increased significantly compared with other groups ( P＜0.01 ).( 5 ) parathyroid hormone ( 1-34 ) in 10-7mol/L,10-8mol/L,10-9 mol/L up-regulated MGP mRNA expression in primary osteoblasts about 6.78,5.31,and 2.23 times than control respectively.It was in a dose-dependent manner.ConclusionThe effect of parathyroid hormone on the expression of matrix gla protein may play an important role in mechanism of postmenopausal osteoporosis

Effect of four different anti-osteoporosis agents on the expression of matrix gla protein in primary osteoblasts of SD rat / 中华内分泌代谢杂志

Objective To observe the expression of matrix gla protein(MGP) mRNA in primary osteoblasts of Sprague-Dawley (SD) rat in vitro after treatment with anti-osteoporosis agents [vitamin K2,PTH,1,25 (OH)2D3,and alendronate],and to investigate the potential role of MGP in the pathogenesis of osteoporosis.Methods Primary osteoblasts(OBs) were derived from sequential trypsin/collagenase-digested calvaria isolated from newborn SD rat (postnastal day 1-3).OBs of the second generation were identified by Van Gieson collagen staining,alkaline phosphatase(ALP) staining and calcified nodules staining.OBs of the fourth generation were selected to interfere with vitamin K2,PTH,1,25 (OH)2D3,and alendronate,then cultured for 24 h in mediums which contained various concentrations of vitamin K2 (10-7,10-6,and 10-5 mol/L),PTH (10-9,10-8,and 10-7 mol/L),1,25 (OH) 2D3(10-10,10-9,and 10-8mol/L),alendronate(10-6,10-5,and 10-4mol/L).After being cultured for 24 h,total RNA was extracted and examined by real-time quantitative RT-PCR.Results The primary cultured cells had typical morphological characters of osteoblast.van Gieson collagen staining,ALP staining,and calcified nodules staining were all positive.Vitamin K2,PTH,1,25 (OH)2D3,and alendronate could modulate the expression of MGP mRNA in osteoblasts in a dose-dependent fashion.MGP mRNA expressions were 2.56-fold,2.12-fold,and 1.57-fold with 10-5,10-6,and 10-7 mol/L of vitamin K2 treatment,respectively.The expressions were 6.78-fold,5.31-fold,and 2.23-fold with 10-7,10-8,and 10-9mol/L of PTH(1-34) treatment,8.93-fold,6.95-fold,and 3.47-fold with 10-8 10-9,and 10-10mol/L of 1,25 (OH)2D3 treatment,and 3.47-fold,2.49-fold,and 1.98-fold with 10-4,10-5,and 10-6mol/L of alendronate treatment.Conclusion Vitamin K2,PTH,1,25 (OH)2D3,and alendronate all canregulate MGP mRNA expression in calvarial osteoblasts in a dose-dependent manner.MGP seems to be a potent target of anti-osteoporosis agents,and involved in the pathogenesis of osteoporosis.

Construction of a 5-Lipoxygenase Transgenic Mice / 中国实验动物学报

Objective To construct a 5-lipoxygenase (5-LO) transgenic mouse model of atherosclerosis.Methods Purified 5-LO fragment was injected into male pronucli and the firtilized eggs were transplanted into pseudopregnant mice.PCR and Southern blot were used to detect the genotype of DNA separated from the newborn mouse tail tissues.RT-PCR and Western blot analysis were used to detect the gene transcription and expression.Results PCR and Southern blot results showed that 7 of 25 mice were transgenic mice.Expression of 5-LO and FLAP was found in the bone marrow,spleen,kidney,and peritoneal cells.Results of RT-PCR and Western blot showed that No.9,20,24transgenic mice expressed a higher level of 5-LO and FLAP than those in the wild type C57BL/6 mice.The expression levels in bone marrow and peritoneal cells were significantly different(P<0.05).Conclusion A 5-LO transgenie mouse line has been established in this study and may be used for future study on the function of 5-LO gene.

Effects of electro-acupuncture on muscle tension and motor function in flaccid paralytic limbs after stroke / 中华物理医学与康复杂志

Objective To investigate the effects of electro-acupuncture on muscular tension and motor function in the flaccid,paralytic limbs of stroke survivors.Methods Seventy-two stroke survivors with flaccid,paralytic limbs were divided randomly into a control group and an electro-acupuncture group.Both grourps received routine therapy and rehabilitation training,but the electro-acupuncture group was also treated with electro-acupuncture.Muscular tension and motor function in the flaccid,paralytic limbs in both groups were evaluated by modified Ashworth scale (MAS)and Fugl-Meyer scores before treatment and at the end of 4 weeks.Results After treatment,in 24 patients of the electro-acupuncture group and 10 patients of the control group muscular tension had increased 1-2 grades.The effectiveness rate(68.57%)in the electro-acupuncture group wag higher significantly than that(27.03%)in the control group(P＜0.01).Fugl-Meyer scores in the electro-acupuncture group were significantlv higher than those in the control group(P＜0.01).Conclusion Electro-acupuncture supplementing traditional rehabilitation improves muscle tension in flaccid,paralytic limbs and promotes the recovery of motor flunction after stroke.

Nursing care of 8 cases of severe hand, foot and mouth disease / 中华护理杂志

This paper reports the nursing care of 8 cases of severe hand,foot and mouth disease,such as first aid measures when admission,airway management during mechanical ventilation,oral care,close observation of vital signs and illness state,brain protection measures,as well as strict disinfection and isolation.As a result,one child died and one gave up the treatment voluntarily.Other six children were discharged with recovery.

Effects of Fluvastatin on microalbuminuria of type 2 diabetes mellitus patients / 中国综合临床

Objective To observe the effect of fluvastatin on urinary albumin excretion rate(UAER)of microalbuminuria in type 2 diabetes patients.Methods Type 2 diabetes patients with mieroalbuminuria were randomly divided into fluvastatin goup(n=126)and control group(n=129).Fluvastatin group was given fluvastatin 40 mg each night.Follow up was conducted for 1.5 years.The UAER and glomerular filtration rate(GFR)as well as blood lipid level before and after therapy were compared.Results UAER in fluvastatin group was significantly lower than that in control group[(59.6±10.5)vs(87.5±12.3)mg/min,P＜0.05]before therapy[(104.4±25.2)vs(110.6±19.7)mg/min,P＜0.05],which was independent of its lowering-lipid effect.But GFR had no signifimicroalbuminuria which is independent of lowering-lipid effect in type 2 diabetic nephropathy patients and delay the progress of diabetic nephropathy.

Preparation of a G418-resistant feeder layer of NIH3T3 cells / 中国组织工程研究

BACKGROUND: Establishment of drug (neomycin or hygromycin) resistant feeder layer cells is necessary for screening the target gene positive clone of embryonic stem cell (ESC) transfected in vitro, and the establishment of drug-resistant NIH3T3 cell line is also important for the screening of other target ESC genes.OBJECTIVE: To obtain a feeder layer of positive cell clone of ESC transfected by pTet-on gene by means of the G418-resistant NIH3T3 cell line establishment.DESIGN: Cell culture and DNA examination.SETTING: Faculty of Laboratory Animal, China Medical University.MATERIALS: NIH3T3 cells were contributed by the Cell Biology Staff Room of China Medical University, and pWL/neo plasmid was a gift from Professor Jin Zhuang of Harvard University Medical College. G418,leukemia inhibitory factor (LIF, ESC GRO 106 U/mL) and DMEM were produced by GIBCO BRL Company, while mitomycin-C (MIT-C), DIG marking and kits were the products of Roche Company. Lipoetin was the product of Invitrogen Company, and bought from Shenyang Lianxing Biotechnology Co.,Ltd.METHODS: ①The pWL/neo plasmid containing neo gene was purified and transfected into NIH3T3 cells by lipoetin.②The transfected cells were further cultured in 25 mL medium containing G418 antibiotic of gradient concentration to observe the survival and apoptosis of cells and measure G418 minimal fatal dose (MFD) to NIH3T3 cells.③The transfected cells were subcultured, and the clone of single cell was selected to 24-pore culture board for screening amplification. At the same time, normal NIH3T3 cells were also selected and added with the selective culture medium at the same dose of G418, as screening negative control. ④The cells were planked with lower cell density, and further screened in DMEM medium containing G418 MFD; Meanwhile, normal NIH3T3 cells were taken as controls, and cultured by G418 contain MFD and by DMEM medium without G418 respectively. Light microscope was used to observe G418R NIH3T3 cells. ⑤G418R NIH3T3 cells and MEF were all applied as the feeder layer to culture the ES-D3 cells, which were observed by light microscope. Cell DNA was prepared, and evaluated by PCR and southern blot.MAIN OUTCOME MEASURES: ①G418 MFD to NIH3T3 cells. ②screening result of G418R NIH3T3 cells. ③growth of G418R NIH3T3 cells. ④growth of ES-D3 cells on G418R NIH3T3 cells.RESULTS: ①G418 MFD to NIH3T3 cells was defined as 500 mg/L.②Under the condition of G418 (500 mg/L), G418R NIH3T3 cell clones were successfully selected. ③The G418R NIH3T3 cells had no difference from normal NIH3T3 cells in morphology and propagation rate.④The ESC cultured in feeder layer present a colony growth, smooth limitation and remained an undifferentiation state. The resistant cell genomic DNA of G418R NIH3T3 cells was assayed with specific neo gene primer, and then neo gene DNA fragment was amplified; Southern blot analysis showed that neo gene fragment integrated into G418R NIH3T3 cells.CONCLUSION: The G418R NIH3T3 cells are established successfully,and the ESC cultured in the G418R NIH3T3 cell feeder layer can keep the characteristics of normal ESC.

Pre-clinical study on tumor vasoactive intestinal peptide receptor scintigraphy / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To develop a tumor imaging agent for vasoactive intestinal peptide (VPAC) receptor and evaluate its biological activity and pharmacokinetics of radiolabeled peptide.</p><p><b>METHODS</b>VIP(28) was modified at the carboxyl terminal by the addition of His-tag which was the chelating site of (99m)Tc(I) and the general purification tag for immobilized metal ion affinity chromatography. Biological activity of the modified VIP(28) analogue MY34 was examined in vitro by radiological cell-binding assay, rabbit internal anal sphincter (IAS) smooth muscle relaxing assay and immunocytochemical stain. The pharmacokinetics of this labeled peptide was examined in C57 mice.</p><p><b>RESULTS</b>MY34 could relax the IAS smooth muscle and bind VPAC receptors on tumor cell membranes. (99m)Tc- MY34, with a yield of about 90%, was stable enough for practical use. Both MY34 and VIP(28) could inhibit the binding between the labeled peptide and VPAC receptor. The pharmacokinetics of [(99m)Tc(H(2)O)(3)(CO)(3)]-MY34 was studied in mice conformed well with the two-compartment model (Wi = 1/C(2)), with a t(1)/(2alpha) of 16.35 min and a t(1)/(2beta) of 1013.56 min.</p><p><b>CONCLUSION</b>MY34 possesses physiological activities and specific receptor binding characteristics similar to those of natural VIP(28).</p>

Antitumor and immunological activities of polysaccharides from rabdosia rubescens / 中国病理生理杂志

AIM: To investigate the antitumor and immunological activities of the polysaccharides obtained from rabdosia rubescens (RRP). METHODS: In vitro and in vivo tumor inhibitory experiments as well as MTT method were used. RESULTS: With increasing concentration, the RRP exerted direct cytotoxicity effect in vitro on the cell growth of EAC mice,and its inhibitory rate in vivo against sarcoma 180 reached 35% ( P