Objective To investigate the effect of insulin-like growth factor- Ⅰ ( IGF- Ⅰ ) on the proliferation and osteogenesis of human periodontal ligament stem cells ( hPDLC ) under three-dimensional (3D) culture system. Methods Human periodontal cells were isolated from the ligament of surgically extracted human teeth, and through the limiting dilution assay, got mono-clone of the cell, hPDLCs were isolated from MesenPRO RS medium. Rotary cell culture system (RCCS) was enrolled to set 3D environment. Control group and experiment groups were assigned according to the concentration of IGF- Ⅰ .There were 5 level of experiment groups (0.1,1,10,50,100 μg/L). Proliferation was tested with methyl thiazolyl tetrazolium ( MTT), and alkine phosphatase (ALP) level was assayed by spectrophotometer to analyze the osteogenesis of hPDLCs. Gene expression of ostetocalcin(OCN)and type Ⅰ collagen (Col Ⅰ )were assayed by reverse transcriptase polymerase chain reaction(RT-PCR). Results In 3D culture system,the effect of IGF- Ⅰ on cell proliferation was significantly different between control group and experiment groups( P ＜ 0.05 ), and there showed significant differences between the group of 0.1 μg/L ( 0.219 ±0.021 ) IGF- Ⅰ and the groups of 50, 100 μg/L(0.287 ±0.011,0.293 ±0.012). However, there showed no significant differences among other groups. Significant differences of ALP activity were observed between the control group and experiment groups, and between the groups of 1, 10 μg/L(0.304 ±0.020, 0.310 ±0.013) and that of 50, 100 μg/L (0.347 ±0.011, 0.344 ±0.010) (P ＜0.05). While no significantdifferences were detected between the group of 1 μg/L and that of 10 μg/L, nor between the group of 50 μg/L and that of 100 μg/L. Expressions of Col Ⅰ and OCN in mRNA and protein level both showed dose-dependent increase. Conclusions In 3D culture system, in the scale of 0.1-100 μg/L, the effect of IGF-Ⅰ on the proliferation of hPDLCs increased dose-dependently. 100 μg/L IGF- Ⅰ promotes osteogenesis of the cells significantly.

Objective To evaluate the effect of different intraoperative insulation methods on hypothermia and surgical site infection in patients with gastrointestinal surgery .Methods A total of 200 patients with gas-trointestinal surgery were randomly divided into control group ( n=100 ) and experiment group ( n =100 ) .The patients in control group were given to cover with quilts.The experiment group patients were given to cover with quilts combined with inflatable warm .The body temperature changes , hypothermia rate , prothrombin time ( PT ) , thrombin time (TT), activated partial thromboplastin time ( APTT), bleeding volume and surgical site infection rate were compared between the two groups . Results In the process of operation , the body temperature of observation group had no obvious change , but compared with the control group , significantly different ( P <0.05 ) .The hypothermia rate of experiment group significantly lower than that of control group ( 10.0%vs 67.0%) , (P<0.05).The PT, APTT and TT of experiment group had no obvious change , statistically different from those of the control group (P<0.05).The bleeding volume of experiment group significantly less than control group ( P<0.05 ) . The surgical site infection rate of experiment group significantly lower than that of control group ( 3.0% vs 17.0%, P <0.05 ) .Conclusion Using inflatable warm nursing on patients with gastrointestinal surgery can effectively maintain body temperature , decrease hypothermia , improve coagulation function , reduce bleeding volume and surgical site infection rate .

Objective To develop an electrochemical impedance immunosensor based on polypyrrole modified microelectrodes for periodontal bacteria rapid quantification.Methods Mcirofluidic chip with embedded IDAM was prepared by conventional photolithography process.Then polypyrrole structure was deposited on microelectrodes through electropolymerization method.Polypyrrole was biofuncationalized with mouse anti-Porphyromonas gingivalis gingipain K IgG antibody using N-(3-dimethylaminopropyl)-N'-ethylcarbodiimide-hydrochloride (EDC) chemistry.The linear relationship between the impedance and bacterial concentration,the impedance characteristic and the feasibility of the developed impedic immunosensor in quantifing Porphyromonas gingivalis was investigated respectively.Results The polypyrrole membrane structure was observed on work electrodes,and the lowest detection limit of the immunosensor was 107 cells/L in pure culture.In the concentration range of 107-1012 cells/L,a linear relationship was found between the normalized impedance change and the logarithmic value of the cell concentration.The total detection time from sampling to measurement was less than 1 h.Conclusions The immunosensor developed in the present study offered some insight into chair-side quantifing periodontal bacteria.

OBJECTIVETo investigate the effect of insulin-like growth factor- I (IGF- I) on the proliferation and alkaline phosphatase (ALP) activity of human periodontal ligament cells (hPDLCs) under three-dimensional (3D) culture system.

METHODSThe hPDLCs were cultured from periodontium of human teeth by the outgrowth method. Rotary cell culture system (RCCS) was enrolled to set 3D culture system. Samples were set to four groups: Negative control group, positive control group (3D group, IGF-I group), and experimental group (3D with IGF- I group). Proliferation was tested with methylthiazolyl tetrazolium (MTT), and ALP activity was assayed by spectrophotometer at 1, 3, 5, 7 d respectively.

RESULTSCompared with that of negative control group, cell proliferation increased significantly in 3D with IGF-I group since 3 d (P < 0.05). Besides, the cell proliferation of 3D with IGF-I group was significantly higher than that of 3D group (P < 0.05). ALP activity of 3D with IGF- I group was significantly higher than that of negative control group, and 3D group at 3, 5, 7 d (P < 0.05).

CONCLUSIONIGF-I significantly promotes the proliferation and ALP activity of hPDLCs under 3D culture system.

Alkaline Phosphatase ; Cell Differentiation ; Cell Proliferation ; Cells, Cultured ; Humans ; Insulin-Like Growth Factor I ; Periodontal Ligament ; Somatomedins

OBJECTIVETo observe the change of cytokine interleukin IL-1 beta, IL-6, IL-10, tumor necrosis factor-alpha (TNF-alpha) occurred in acute paraquat (PQ) poisoning rats and to investigate the mechanism of acute lung injury caused by paraquat (PQ) poisoning.

METHODSAll 72 healthy adult Wistar rats were random assigned into normal control groups, paraquat high dose group (120 mg/kg), paraquat middle dose (60 mg/kg) group, paraquat low dose group (30 mg/kg). Three observing periods of time included 8, 24, 72 h and the standards of TNF-alpha, IL-1 beta, IL-6, IL-10 were determined.

RESULTSEvery index of the PQ group was significantly higher than that in the NS group at the same period of time (P<0.05 or P<0.01). In the 72 h group, the high dose group was significantly higher than the middle and low dose group (P<0.05), and there was no significantly difference between the middle and low dose group (P>0.05). For the comparison of index in the same dose group, the group of 72 h was much higher than 8 h group and 24 h group (P<0.05), and there was no difference between the 8h group and 24 h group (P>0.05).

CONCLUSIONThe cytokine may play an important role in paraquat-induced acute lung tissue injury.

Acute Disease ; Animals ; Cytokines ; blood ; Disease Models, Animal ; Female ; Interleukin-10 ; blood ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Male ; Paraquat ; poisoning ; Rats ; Rats, Wistar ; Tumor Necrosis Factor-alpha ; blood

AIMTo approach the relationship between lung injury induced by shock/reperfusion and nitric oxide as well as the beneficial effect of taurine.

METHODSTwenty four rabbits were divided randomly into 3 groups (n = 8): control group, shock group, taurine group. The model of lung injury induced by shock/reperfusion was used. The activities of nitric oxide synthase (NOS), superoxide dismutase (SOD), the contents of malondialdehyde (MDA), nitric oxide products (NO2-/NO3-) in plasma and lung homogenate, lung wet/dry weight, lung water content, lung permeability index, and protein content in the pulmonary alveolar lavage fluid were measured. Meanwhile, pathologic samples treated routinely.

RESULTS(1) At 3 hours after reperfusion, the activities of SOD in plasma and lung homogenate decreased markedly, but the other indexes above mentioned were increased significantly compared with the control group (P < 0.01). (2) A close correlation was shown between MDA content and NO2-/NO3- content in plasma and lung. Furthermore, the content of NO2-/NQ3- in lung homogenate showed strong positive correlation with the lung injury parameters. (3) Taurine (40 mg x kg(-1) i.v.) could attenuate all the changes above mentioned at the same time points of reperfusion.

CONCLUSIONNO may play an important role in lung injury induced by shock/reperfusion. Taurine can ameliorate the lung injury, mechanism of which may be related to decreasing the generation of NO and anti-lipoperoxidation.

Animals ; Lung ; drug effects ; metabolism ; Lung Injury ; etiology ; prevention & control ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Rabbits ; Reperfusion Injury ; complications ; drug therapy ; Shock, Hemorrhagic ; complications ; drug therapy ; Superoxide Dismutase ; metabolism ; Taurine ; pharmacology ; therapeutic use

AIMTo investigate the effect of taurine on nitric oxide synthase (NOS) activity and nitric oxide products (NO2 /NO3 ) content in myocardium and plasma during shock resuscitation.

METHODSTwenty-four rabbits were divided randomly into 3 groups (n=8): control group, shock group, taurine group. The model of hemorrhagic shock resuscitation was used. The activities of nitric oxide synthase (NOS), lactate dehydrogenase (LDH) and the contents of nitric oxide products (NO2- /NO3-) in plasma were observed before shock and shock 1.5 hours, after resuscitation 1 hour, 2 hours and 3 hours. The activities of NOS and the contents of NO2-/NO3- in myocardium homogenate were measured after resuscitation 3 hours. Meanwhile, pathologic samples treated routinely.

RESULTS(1) During resuscitation, the activities of NOS, LDH and the contents of NO2- /NO3- in plasma of shock group were significantly higher than that of before shock and shock 1.5 hours (P < 0.01). (2) After resuscitation 3 hours, the activity of NOS and the contents of NO2- / NO3 in myocardium of shock group were significantly higher than that of control group (P < 0.01). The cardiac myocyte appeared edema, fatty degeneration. (3) All the changes of above mentioned could be attenuated by intravenous injection taurine (40 mg/kg) (P < 0.01).

CONCLUSIONThese results suggest that the NOS activation and NO release may mediated myocardium injury induced by shock resuscitation, taurine can ameliorate the myocardium injury, which may be related to decreasing the generation of NO.

Animals ; Myocardium ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Plasma ; metabolism ; Rabbits ; Resuscitation ; Shock, Hemorrhagic ; blood ; metabolism ; Taurine ; pharmacology

OBJECTIVETo clone novel gene from suppression subtraction library established for screening down-regulated genes in gastric carcinoma, and the effects of novel gene on gastric tumorigenicity were analyzed.

METHODSSequencing results of 860 positive colonies chosen randomly were compared by Blast program in GenBank. Novel gene fragment was amplified by rapid amplification of cDNA ends (RACE). The mRNA expression of novel gene was detected by Northern blot and semi-quantitative PCR in 25 cases of gastric carcinoma tissue and counterpart normal gastric mucosa. The structure and chromosomal location of novel gene were investigated by Bio-message technique.

RESULTSA 233 bp novel gene fragment was screened out from 860 clones and a 802 bp novel gene was obtained by RACE. The novel gene was named as GDDM, registered in the number of AF494508 by GenBank. The mRNA expression of GDDM in gastric carcinoma tissue (4.496+/-0.637) was significantly lower than that in the counterpart normal gastric mucosa (36.919+/-6.290)(P<0.01). Chromosomal location of GDDM gene was at 4q31.

CONCLUSIONThe cloned novel gene, GDDM, is down-regulated in gastric carcinoma, and it is likely to be involved in gastric tumorigenicity.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; Down-Regulation ; Gene Amplification ; Gene Expression Regulation, Neoplastic ; Gene Library ; Genes, Neoplasm ; Humans ; Molecular Sequence Data ; Stomach Neoplasms ; genetics ; metabolism

OBJECTIVETo investigate the feasibility and efficacy of neoadjuvant chemoradiotherapy followed by combined thoracoscopic and laparoscopic esophagectomy (CTLE) in the treatment of advanced esophageal carcinoma.

METHODSFrom June 2011 to February 2012, 11 patients with locally advanced esophageal carcinoma underwent neoadjuvant chemoradiotherapy followed by CTLE (clinical stage IIB-IIIA). NP (vinorelbine pin and cisplatin) or TP (program paclitaxel-pin and cisplatin) were applied as preoperative chemotherapy. During the same period, conventional fractionated radiotherapy was used with the radiation dose of 40 Gy/20 F. At four to six weeks after CRT, 11 patients received three-incision CTLE.

RESULTSDuring chemoradiation, 9 patients developed bone marrow suppression. The interval between completion of chemoradiation and surgery was (49.6±15.4) d. Intraoperative findings revealed local fibrosis in one patient (75 days after chemoradiation) while operative difficulty was not increased in the remaining 10 patients. Compared to 15 patients who received surgery alone, operative time was shorter [(242.3±27.0) min vs.(280.5±27.2) min, P=0.002] and intraoperative blood loss was less [(168.2±95.6) ml vs. (244.5±84.8) ml, P=0.042], the number of removal lymph nodes was similar [(19.5±5.8) vs. (20.5±7.1), P=0.683], postoperative hospital stay was prolonged [(18.9±10.3) d vs. (12.5±4.6) d, P=0.020]. The postoperative complication rate was 36.4% including cervical anastomotic leak with pulmonary infection (n=1), cervical anastomotic fistula and hoarseness (n=1), pulmonary infection with pleural effusion (n=2). Follow up ranged from 1 to 9 months, and no recurrence was found.

CONCLUSIONThe neoadjuvant chemoradiotherapy followed by combined thoracoscopic and laparoscopic esophagectomy in the treatment of locally advanced esophageal carcinoma is safe, feasible, and the short-term outcomes are favorable.

Adult ; Aged ; Esophageal Neoplasms ; surgery ; therapy ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Middle Aged ; Neoadjuvant Therapy ; Preoperative Care ; Thoracoscopy ; Treatment Outcome

BACKGROUNDLittle attention has been paid to the expression of heat shock protein 27 (HSP27) in patients with reflux esophagitis (RE), and few studies of the importance of HSP27 in esophagitis have been carried out in animal models. This study aimed to explore the expression of HSP27 in the esophageal tissue of rats with RE.

METHODSEighty female Wistar rats were randomly divided into experimental groups A and B and control groups C and D (n = 20 in each group). To establish RE, rats in the two experimental groups received pylorus and forestomach ligations, while rats in the control group received gastrostomy and gastric perforation repair. The rats in groups A and C were sacrificed 7 days after surgery, and the rats in groups B and D were sacrificed 14 days after surgery. In groups A and B, 10 and 8 rats were diagnosed with RE by pathological examination, respectively (they were included in groups A' and B', respectively). The histopathological diagnosis of all the lower esophageal tissues in groups C and D was normal and 20 normal specimens were randomly selected for groups C' and D' with 10 specimens in each group. Macroscopic and microscopic esophagitis scores were assessed for the specimens in groups A' and B'. Lower esophageal tissues were collected from groups A', B', C', and D', and paraffin-embedded slices were made using part of the tissues. The expression of HSP27 in the tissues was detected using the two-step streptavidin-peroxidase immunohistochemical method. Some collected tissues were frozen, and expressions of HSP27 mRNA were detected using fluorescence quantitative polymerase chain reaction (FQ-PCR).

RESULTSMedian macroscopic and microscopic esophagitis scores in groups A' (n = 10) and B' (n = 8) were 1.0 and 1.5, and 2.0 and 2.5, respectively. There were no significant differences in the macroscopic or microscopic esophagitis scores between the two groups (Z = -0.330, P = 0.741; Z = -0.142, P = 0.887, respectively). Immunohistochemical staining showed that HSP27 was expressed in all layers of the esophageal epithelia in RE and control rats. FQ-PCR showed that HSP27 mRNA levels in the lower esophageal tissue in RE group (groups A' and B') were higher than those in control group (groups C' and D') (Z = -0.249, P = 0.001). HSP27 mRNA expression in the lower esophageal tissue was significantly different in groups B' and D' (Z = -3.027, P = 0.002). And the levels of HSP27 mRNA expression in severe RE group (microscopic esophagitis score: 3) were higher than in mild RE group (microscopic esophagitis score: 1-2) and control group (Z = -3.396, P = 0.001; Z = -3.855, P < 0.001).

CONCLUSIONSHSP27 mRNA expression in the lower esophageal tissue of rats with RE is significantly higher than in the normal controls. Although reflux is a persistent stimulating factor, increased expression of HSP27 in the lower esophageal tissue of rats with RE requires aggravated esophageal injury.

Animals ; Esophagitis, Peptic ; metabolism ; Esophagus ; metabolism ; pathology ; Female ; HSP27 Heat-Shock Proteins ; genetics ; metabolism ; Immunohistochemistry ; Polymerase Chain Reaction ; Rats ; Rats, Wistar