OBJECTIVERNA interference was applied to knockdown the Dhcr7 gene in mouse embryonic palatal shelves to facilitate understanding of the function of Dhcr7 gene variants in the fusion of palatal shelves.

METHODSThe pAdTrack-CMV-siDhcr7 was constructed using the specific siRNA sequence of Dhcr7 from C57BL/6J mouse. The pAdTrack-CMV- siDhcr7 of positive clones was reconstructed in vitro, and the recombinant adenovirus pAdEasy-1-siDhcr7 of kanamycin resistance was screened. The adenovirus vector DNA was then prepared for transfecting the embryonic palatal shelves. Thirty pairs of embryonic palatal shelves at 13.5 d gestational age were harvested and then randomly divided into the following three groups: normal control group (n = 10), which included palatal shelves inculture medium without cholesterol; blank adenovirus control group (n = 10), which included palatal shelves in culture medium without cholesterol and blank adenovirus; and experimental group (n = 10), which included palatal shelves in culture medium without cholesterol and adenovirus encoding Dhcr7 siRNA. At 48 h after in vitro cultivation, the mRNA and protein of the palatal shelves were obtained for scanning electron microscopy (SEM), reverse transcription polymerase chain reaction (RT-PCR), and Western blot analyses.

RESULTSSEM showed that the palatal shelves of the normal control and blank adenovirus control groups fused and formed continuous palates, whereas those of the experimental group was almost undeveloped but exhibited large gaps between the two palatal shelves. RT-PCR and Western blot analyses showed that the mRNA and protein of Dhcr7 in the experimental group decreased compared with those in the normal control group with a significant difference (P < 0.05).

CONCLUSIONResults indicate that Dhcr7 gene silencing affects the fusion of palatal shelves. Thus, Dhcr7 gene may serve a function in the normal development of palates.

Animals ; Cleft Palate ; Gene Silencing ; Mice ; Mice, Inbred C57BL ; Microscopy, Electron, Scanning ; Organ Culture Techniques ; Oxidoreductases Acting on CH-CH Group Donors ; Palate ; growth & development ; RNA, Messenger

Objective To explore the relationship of the presence and severity of anxiety and depression with the increased inflammatory activity,as marked by the serum levels of high sensitivity C reactive protein(hs-CRP)after acute coronary syndrome(ACS).Methods Serum hs-CRP levels were measured in 647 ACS patients within 36 hours after onset of event.Depression and anxiety were evaluated by self-reporting standardized questionnaire,using a validated Chinese version of Hospital Anxiety and Depression Scale(HADS)(14 items)within 7 days.Results In ACS patients,serum levels of hs-CRP(mg/L)were lower in those with anxiety than those in control group[(10.43?3.55)mg/L vs(13.19?4.90)mg/L,P0.05].Conclusion Presence and severity of depression is associated with increased activity of inflammation in patients with ACS;however,anxiety does not have such an association with inflammation in patients with ACS.

Objective: To investigate the predictive value of an early inflammatory response factor, Rho kinase activity for left ventricle remodeling (LVR) in patients with acute ST-segment elevation myocardial infarction (STEMI).
Methods: A total of 120 acute STEMI patients treated in our hospital from 2010-10 to 2013-06 were studied, all patients were ifrst time received primary percutaneous coronary intervention (PCI) with stent implantation. Rho kinase activity and B-type natriuretic peptide (BNP) were measured before PCI, echocardiography was conducted at 24 hours and 12 months after STEMI respectively to clarify LVR diagnosis. The patients were divided into 2 groups as LVR group, n=97 and Non-LVR group, n=23, the above indexes were compared between 2 groups.
Results: The level of Rho kinase was higher in LVR group than that in Non-LVR group, P<0.001, after adjustment, Rho kinase was the independent predictor for LVR (OR 3.36, 95%CI 2.01–5.78, P<0.001). The ROC of Rho kinase was 0.88 (95%CI 0.82–0.94) and the ROC of BNP was 0.54 (95%CI 0.41–0.70).
Conclusion: High Rho kinase activity could predict LVR in acute STEMI patients with primary PCI and stent implantation.

BACKGROUND:Recombinant bovine basic fibroblast growth factor is a manifold effect cytokine which can promote angiogenesis, wound healing, tissue repair and bone regeneration. Recombinant bovine basic fibroblast growth factor with good histocompatibility is easy to operate and has been widely used in oral and maxil ary surgery.
OBJECTIVE:To evaluate the effect of recombinant bovine basic fibroblast growth factor against dry socket syndrome after tooth extraction.
METHODS:A total of 160 patients who had been extracted mandibular third molar were selected and randomly divided into two groups. In the experimental group, recombinant bovine basic fibroblast growth factor was put into the sockets after mandibular third molars were extracted, while in the control group, we let the wounds to be healed natural y without any materials. The incidence of dry socket syndrome was observed and compared between two groups at 3 days, 5 days and 1 week after tooth extraction.
RESULTS AND CONCLUSION:One patient had dry socket after operation in the experimental group, and the incidence was 1.25%. In the control group, 10 patients suffered from dry socket, and the incidence was 12.5%. There was a significant difference in the incidence of dry socket between the two groups (P<0.01). There was visible granulation tissue within the tooth socket after tooth extraction in the experimental group, and extraction sockets narrowed and were fil ed with granulation tissues, which was 1-2 days earlier than the control group. No al ergies, tissue hyperplasia and other local and systemic reactions occurred in patients receiving implantation of recombinant bovine basic fibroblast growth factor gel. These findings indicate that local implantation of recombinant bovine basic fibroblast growth factor gel after mandibular tooth extractions can speed up the healing of dental extraction wounds.

Objective To evaluate the safety,tumor radical and early postoperative efficacy through comparison of laparoscopic gastric D2 radical surgery with traditional open gastric D2 radical surgery.Methods 254 patients with upper stomach cancer underwent surgery were selected,132 cases using conventional open gastrectomy (the traditional laparotomy group),122 patients underwent laparoscopic radical gastrectomy (laparoscopic surgery group).Laparoscopic surgery group with traditional open surgery group had no statistically significant differences in gender,age,tumor location,histological type and TNM staging.Results Open surgery group and laparoscopic surgery group had statistically significant differences in operative time [(235.78±31.56) min,(256.43±54.08) min,P ＜ 0.001],blood loss [(326.69±89.73) ml,(158.31±62.98) ml,P ＜ 0.001],incision length [(16.53±2.34) cm,(5.51±1.15) cm,P ＜ 0.001],gastrointestinal recovery time [(4.22±0.91) d,(3.31±0.83) d,P ＜ 0.001],first time eating liquid [(5.78±0.95) d,(5.56±0.78) d,P ＜ 0.001] and postoperative hospital stay [(12.62±2.89) d,(11.18±1.78) d,P ＜ 0.001].The total number of lymph node dissection and complications was not statistically significant.Conclusions Laparoscopic gastric D2 radical surgery is a safe,minimally invasive surgical method.Laparoscopic gastric D2 radical surgery has the same lymph node dissection and good early outcome compared with the traditional gastric D2 radical surgery,but postoperative recovery fast and less invasive.

Objective To investigate the effect of mitogen-activated protein kinase(MAPK)pathway on the transcriptional expression of mdr1 gene induced by doxorubicin ( DOX)and study the transcription regulation of mdr1 gene.Methods K562 cells were treated with DOX(0.01 μg/ml)with the initial concentration of 0.01 μg/ml for 24 hours,then change the culture media without DOX.K562 cells were cultured until the its status wag recovered.Subsequently the cells were treated with DOX(0.02μg/ml)for 24 hours again.The concentration of DOX was increaged until 0.05 μg/ml by following the protocol above.K562 cells were collected at the concentration of 0.01 μg/ml,0.03μg/ml and 0.05μS/ml DOX.Expression of mdr1 gene were examined by reverse transcription-polymerase chain reaction(RT-PCR).Pglycoprotein(P-gP)wag detected by flow cytometry.Western blot wag performed to detect ERK and P-ERk.K562 cells were pretreated with MAPK inhibitor PD98059 for 1 hour.and then DOX was added.RT-PCR and FCM were used to detect the expression of mdr1 mRNA and P-gp.Results When K562 cells were exposured to DOX.the phosphorylation of ERK wag increaged.the mdr1 gene wag highly expressed as well as its corresponding protein P-gp.When the concentration of DOX was 0.05μg/ml,the expression of mdr1 gene and P-gp were increased over 5 fold.When K562 cells were pretreated with MAPK inhibitor PD98059,the expression of mdr1 gene induced by DOX(the concentration was 0.03 μg/ml and 0.05 μg/m1)was effectively inhibited by(74.1±0.11)%and(70.2±0.14)%respectively.Conclusions DOX could induce the expression of mdr1 gene in K562 cells accompanied by the activation of MAPK/ERK pathway.The block of activation of ERK could inhibit the induced expression of mdr1 gene.

AIM: In order to study the relationship between mitochondrial deficiency and Alzheimer's disease(AD), we used sodium azide, a specific inhibitor of cytochrome C oxidase(COX), to develop a cell model of mitochondrial complex IV deficiency and investigated the impairment of microtubules and microtubule-associated proteins. METHODS: Primary cultured hippocampal neurons of hewborn rats were exposed to sodium azide ,then cell viability was measured by MTT method; cell morphology, immunofluorecence-stained cellular microtubules and microtubule-associated proteins were observed by confocal microscopy. RESULTS: Primary cultured hippocampal neurons were exposed to 8-128 mmol/L sodium azide for 3-24 h, MTT absorbance decreased dose-and time-dependently. Exposed to 64 mmol/L sodium azide for 6 h, the processes of cells retracted, synapses disappeared, axons were shortened under contrast microscope. Meanwhile, microtubles were disassembled and became disorderly, the expression of microtubule-associated proteins were also reduced especially in the processes observed by confocal microscopy. CONCLUSIONS: Sodium azide inhibits the assembly and polymerization of tubulin in microtubules which may be reduced by low expression of microtubule-associated proteins in nerve cells. The damage of axons induced by microtubule collapse further blocks the intercellular signal transduction and intracellular material transportation which are important causes in cell death.

Objective To explore the occurrence and development of schistosomiasis japonica hepatic fibrosis, and the treatment countermeasure. Methods Three groups of patients,including advanced schistosomiasis (splenectomy involved), non-advanced schistosomiasis splenomegaly and advanced schistosomiasis with hepatitis, were followed up. Biochemical tests including hepatic fibrosis indexes, liver ultrasonography,lymphocyte subcluster, and membrane CD35 were examined. Immunohistochemical tests of liver cells were performed in some patients. Results A total of 94.06% of 1212 patients took various anti-schistosome treatments. The difference was significant for positive antigen and antibody to schistosome, LN and HA ,and the ultrasonic indexes of liver fibrosis among the three groups. The degree of liver substance pathohistological examinations showed inflammation had a positive correlation with hepatic fibrosis. The contents of C-Ⅰ, C-Ⅲ in portal regions, central vein and hepatic sinusoid were increased in advanced schistosomiasis hepatic fibrosis. In hepatic fibrosis, hepatic sinusoid showed two pathological changes of dilation and stricture. Conclusions In schistosomiasis japonica, hepatic sinusoid changes play a great role in hepatic fibrosis.Hepatic fibrosis would still develop even after the thorough anti-schistosome therapy. So, anti-fibrosis therapy is necessary.

Objective To study the long-term efficacy of splenectomy for patients with advanced shistosomiasis japonica.Methods Levels of WBC,RBC,PLT,EOS,ALT,ALP,GGT,A,TB,HA,LN,Ⅳ-C,PCⅢ,IGG,IGA,C3,C4,CD3,CD4,CD8,CD19 in periphetral venous blood were determined in 239 patients with advanced shistosomiasis.Meanwhile,the liver,gallbladder and spleen were examined with ultrasonography.Results The levels of WBC,PLT,EOS,ALT,ALP,IGG,IGA,LN,Ⅳ-C,CD19 increased in splenectomy group,the levels of A,TB,CD3,CD4,C3,C4 decreased in splenectomy group,while RBC,HA,PCⅢ,CD8 were not changed.Conclusion Splenectomy is a danger to hepatic function.Humoral immunity increases while cellular immunity decreases in splenectomy group.Splenectomy may aggravate the hepatic fibrosis in patients with advanced shistosomiasis.

Objective To observe and assess the effect of different dosages of aspirin on inflammatory biomarkers, hemorheology (platelet aggregation rate) and clinical prognosis in patients with acute coronary syndrome ( ACS). Methods ACS patients were randomly assigned to receive different dosages of aspirin treatment orally. Patients in group A,B and C took 100 mg, 500 mg and 1000 mg of aspirin per day respectively. They were treated and followed-up for 1 year. High-sensitivity C-reactive protein ( hsCRP) , IL-6, tumor necrosis TNFot and platelet aggregation rate were examined and major adverse cardiac events( MACE) were recorded. Results A total of 312 patients with ACS were enrolled in the study. The baseline characteristics of the three groups were not different with respect to age, gender, cardiovascular risk profile, level of inflammatory biomarkers and concomitant treatment before and after randomization. The levels of baseline serum hsCRP, IL-6 and TNFa were higher in subjects of the study as compared with normal reference value (P<0. 05, <0. 05, <0. 01) and they decreased significantly after therapy with 3 different doses of aspirin (detected at 30 days, 6 months and 12 months, P <0. 001 ) , but there were no significant differences among the three groups( P >0. 05) . Rehospitalization , MACE and the change of platelet aggregation ratio were not significantly different among the three groups. The incidence of gastrointestinal complaints was significantly higher in groups B and C than in group A ( P < 0. 05 ). Conclusions The levels of serum inflammatory biomarker increase in patients with ACS. Aspirin therapy may decrease the level of inflammatory markers significantly, but increasing the dosage of aspirin from 100 mg to 1000 mg daily does not decrease the level of inflammatory markers and the clinical MACEs further. However, the incidence of gastrointestinal complaints increase significantly with the increase of aspirin dosage.