133 teeth in 34 children aged 4 to 8 year-old were recruited for the treatment of bilateral maxillary primary canines or primary molars with deep caries,chronic pulpitis or chronic periapical periodontitis under local anesthesia.Buccal infiltration(BI)with conventional syringe(CS)was used for one quadrant,anterior middle superior alveolar anesthesia(AMSA)with computer-controlled local anesthesia de-livery system(C-CLADS)on the contralateral side.The injection duration(s)of CS and C-CLADS was 63.6 ±22.6 and 136.6 ±12.4 re-spectively(P =0.000).The pain perception of C-CLADS injection was significantly lower than that of CS(P <0.05),that during operation was not statistically different(P >0.05).55.9% children preferred C-CLADS anesthesia.

The relationship between diabetes and osteoporosis has gradually become a new research hotspot , but is still much controversy .High concentration of blood glucose can be bond to varieties of proteins in vivo and form glycosylation end products in col -lagen tissue and reduce bone strength , besides it can inhibit RANKL induced osteoclast formation and bone resorption by repressing redox-sensitive NF-kappa B activity .A series of diabetes mellitus complications can induce less local bone nutrition supply and calcium and phosphorus sorption , reduce muscle tension and bone mineral density , which leads to the increase of fracture risk .Type 2 diabetes patients with overweight or obesity often have increased secretion of inflammation factors , adiponectin and leptin , which promotes the formation of osteoclasts and encourages bone marrow stromal stem cells to differentiate into adipocytes ;On the other hand , adiponectin and leptin can facilitate osteoblasts proliferation , differentiation and activity , and promotes bone formation .This article will discuss how diabetes works on bone metabolism and the underlying pathophysiological and physiological mechanism .

Objective In order to study the important role of two specific VEGF receptors(Flt 1 and KDR) in angiogenesis, we design the experiment to detect the gene expression of Flt 1 and KDR in human umbilical vein endothelial cells cultured in hypoxium. Methods We cultured the human umbilical vein endothelial cells for different time and treated it with vascular activate factor, such as endothelin (ET), Nitric oxide (NO), LNNA (inhibitor of NO). The expressive level was analyzed by Northern blot or RT PCR. Results Hypoxia increases flt 1 mRNA in endothelial cells at 6 hours. Administration of ET stimulated an increase in flt 1 mRNA. NO inhibited expression of flt 1 mRNA. LNNA increased its expression. Hypoxia partly affected the expression of KDR. LNNA enhanced the VEGF transcripts. Conclusion Hypoxia up regulated flt 1 gene expression, partly KDR. ET enhanced and NO inhibited flt 1 gene expression.

Objective To find a indicator in urine to assist diagnosis of cerebral infarction,we investigated the changes of urine ferritin in patients with cerebral infarction.Methods Collected serum from 30 healthy volunteers and 53 patients with cerebral infarction(CI),with ratio of males to females 19∶11 vs.30∶ 23,and average age was (56 ± 11) vs.(63 ± 11) respectively.Collected urine from 39 heathy volunteers and 33 patients with CI,with ratio of males to females 25∶14 vs.20∶ 13,and average age was (57 ± 10) vs.(64 ± 11) respectively.Ferritin concentration in all samples was detected with the chemical luminescence method,and the urine ferritin/urine cretinin (uFer/uCr) ratio was calculated.Differences of serum ferritin and uFer/uCr between healthy volunteers and CI patients were analyzed with the statistical software under Wilcoxon test.Urinary proteins were extracted with absolute ethanol.The extracted ferritin was displayed with Western blot for light chains and heavy chains to investigate the difference between healthy volunteers and CI patients.Results uFer/uCr was higher than that from healthy volunteers (U =292.00,P ＜0.05),and the median was (14.86 vs.6.22) μg/g.Moreover,after extraction of urinary proteins with alcohol,both light chains and heavy chains of ferritin in urine from CI patients were much more obvious than that from healthy volunteers.Conclusion The uFer/uCr ratio could provide a new proof to assist the diagnosis of cerebral infarction.

Objective To investigate the regulation and expression of VEGF gene in human umbilical vein endothelial cells under hypoxia and the influence of vascular activators,such as endothelin(ET),nitric oxide(NO)and NO synthesis inhibitor N-nitro-L-Arginie(L-NNA),on the expression of VEGF.Methods Culture the human umbilical vein endothelial cells and then treat by hypoxia and vascular activators.The expression on VEGF mRNA level and protein level were assayed by northern blots,ELISA and computer figure analysis system respectively.Results When human umbilical vein endothelial cells were subjected to hypoxia for 6 hours,the expression of VEGF began to increase.ET could increase the expression and NO inhibited it,and LNNA affected the expression of the VEGF.VEGF protein concentration in the cell culture media matches the mRNA result.They are 6 hours hypoxia (8^2?1.1)?g/L,ET+6 hours hypoxia(9^37?1^02)?g/L,NO+6 hours hypoxia(2^86?0^91)?g/L,LNNA+6 hours hypoxia(14^75?1^87)?g/L respectively.Conclusion (1)Hypoxia can induce the expression of VEGF in human umbilical vein endothelial cells.(2)ET can increase the inducible effect of hypoxia,NO can inhibit it and LNNA stimulated the expression of VEGF in human umbilical vein endothelial cells.

AIM: To study the application of macular ganglion cell complex (mGCC) and peripapillary retinal nerve fiber layer thickness (pRNFL) measured by optical coherence tomography (OCT) in the early diagnosis of glaucoma.METHODS: Case-control study.Eighty-six subjects, including 30 eyes in normal subjects, 27 eyes in suspected primary open angle glaucoma, 29 eyes in primary open angle glaucoma were enrolled in this study.The thickness of mGCC and pRNFL were measured by OCT.The area under the receiver operating characteristic(AROC) curve at fixed specificities were calculated for each parameter.RESULTS: There were significant differences in mean pRNFL thickness, superior pRNFL thickness and inferior pRNFL thickness between normal group, suspected glaucoma group and early glaucoma group (P=0.001, 0.004, 0.011).The mean mGCC thickness, the thickness of the top mGCC, the thickness of the lower mGCC were statistically significant (P=0.008, 0.002, 0.003);the difference of general loss of volume (GLV) and focal loss of volume (FLV) between the three groups was statistically significant (P=0.002).Compared with the normal group, all the pRNFL and the mGCC parameters were higher in the suspected glaucoma group, and the FLV had the highest AROC (0.801), all the remaining AROC was >0.700 except above Prnlf (0.688).Compared with the normal group and the early glaucoma group, all the pRNFL and the mGCC had higher AROC, average mGCC was hightest(0.804), all parameters AROC were >0.700 except mean pRNFL (0.683).In suspected glaucoma group, 58% patients had abnormal mGCC thickness and 23% had abnormal pRNFL thickness;in early glaucoma group, 98%patients had abnormal mGCC thickness and 90% had abnormal pRNFL thickness;in normal group, 93%patients had abnormal mGCC thickness and 93%had abnormal pRNFL thickness, the correlation between the three groups was statistically significant (x2=12.11, P<0.05).CONCLUSION: OCT measurement of mGCC thickness and pRNFL thickness in early glaucoma have good diagnostic ability;mGCC thickness measurement can be used as an effective method for early diagnosis of glaucoma.

OBJECTIVE: To evaluate the quality of Gentiana scabra in Qingre jiedu oral solution. METHODS: HPLC method was established to determine the content of gentiamarin in the oral solution. 299 batches of Qingre jiedu oral solution in circulation were measured and evaluated. RESULTS: Developed HPLC method is simple and reliable. The quality of Qingre jiedu oral solution in circulation is not satisfied. CONCLUSION: Great importance should be attached to the manufacture procedures monitoring of Qingre jiedu oral solution and quality control of G. scabra.

Objective To discuss the clinical efficacy of point-injection with nerve growth factor (NGF) in treatment of infantile cerebral palsy.Methods According to the time order,50 cases with cerebral palsy were randomly divided into the treatment group(30 cases) and the control group (20 cases).Both groups were given the conventional treatment of Bobath mainly exercise therapy,homework therapy and the language therapy.In addition,the treatment group was given the treatment of point-injection with NGF.Gesell scale was used to evaluate all the patients before and after treatment.Results After treatment,the Gesell evaluation of the treatment group was higher than control group,and the difference was statistically significant (t =-2.096,-2.187,-2.048,-2.053,-2.225,all P ＜0.05).Conclusion On the basis of rehabilitation training,point-injection with NGF could improve intelligence,motor function and language in children with cerebral palsy,and the efficacy was obvious.

ObjectiveTo facilitate the establishment of fully automatic clinical diagnostic nucleic acid assay,a study was carried out for developing a fast and convenient method of extracting highly purified viral nucleic acid from human blood with magnetic beads.Method Five KHB (Shanghai Kehua Biological Engineering Company Limited) internal strong positive HBV serum samples were used in the process,for the method development,screening and selecting the magnetic beads based on magnetism,suspension power and extraction effects of HBV DNA,the optimization of the selected key components of extraction reagents were performed,which including the quantity of magnetic beads( 1 - 1.75 μl/ml),the buffer pH of lysis (4 -9),washing conditions with or without alcohol,elution for proliferation with or without magnetic beads,and etc.The evaluation for the extraction method was carried out including linearity,precision,sensitivity,specificity,anti-interference and resistance to contamination.The magnetic beads method was compared with domestic mature boiling method in a quantitative test with 100 HBV serum samples,the magnetic beads method was also compared with the interntional Gold standard Roche COBAS Tagman test method ( Roche method) in a quantitative test with 50 HBV serum samples and the results were statistically compared for their linear regression.ResultsOptimization experiment result showed superparamagnetic beads A was the best among the candidates ( C,B,D) in terms of their physical properties and extraction effects of DNA.The optimum condition of the assay were as follow:magnetic beads best used at 1.25 μl/ml,buffer pH 7 - 8 of lysis,ethanol was eliminated from washing buffer,proliferation can be carried out with magnetic beads.The parallel comparison experiments with five KHB internal strong positive HBV serum sample and gradient diluted serum sample between beads method and boiling method were carried out.The results showed that the linear range of beads method was 4 × 101 -4 × 108 IU/ml and that of boiling method was 4 × 102 -4 × 108 IU/ml.The sensitivity for beads method was 30 IU/ml and that of boiling method was 100 IU/ml.The precision CV of beads method was 6.0％ and boiling method was 11.9％.The false positive rate was 0 for beads method vs 3.1％ for boiling method.The anti-interference ability experiment showed that when sample contained sodium heparin concentration exceed 62.5 U/ml, there was significant drop of HBV DNA detection with boiling method and no HBV DNA was detected when sodium heparin concentration in the sample reached 1000 U/ml.However,sample sodium heparin concentration had no effect on HBV DNA detection with beads method.The comparison study between beads method and boiling method was carried out by use of 100 clinical HBV positive serum samples.The result showed that the sensitivity for beads method was 86％ vs boiling method 74％.The correlation between the two method was R2 =0.948,P ＜ 0.05.The comparison study between beads method and Roche method was carried use of 50 clinical HBV DNA positive serum samples and the result showed that sensitivity for beads method was 95％ (49/50) vs Roche method was 100％ (50/50).The correlation was R2 =0.963,P ＜ 0.05.ConclusionsThe established method for automatic nucleic acid extraction with magnetic bead displayed characteristic of high yield nucleic acid extraction,broader testing range,accurate quantification and convenient in operation.This can be one choice for nucleic acid extraction and applicable for clinical automatic diagnostic system.( Chin J Lab Med,2012,35:843 -850 )

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