Targeting the key questions in the scale development process of TCM syndrome, such as the definition of the concept, the construction of the theoretical framework, the quantitative classification of the items, the rational use of the statistical methods and so on, this article put forward that on the basis of carefully distinguishing the three concepts of syndrome diagnosis, syndrome evaluation and disease diagnosis, and based on TCM dialectical thinking and mathematical validation to build theoretical framework. A scientific and reasonable quantitative classification method was established based on the reliability and validity as indexes. Non-linear intelligent mathematical statistics and symptomatic index groups were used to analyze the ideas and methods of data mining, with a purpose to improve and perfect the methodology of the development of syndromes scale and to improve the establishment and application of the syndrome scale.

Background Neurotransmitter secretion disorder induced by chronic manganese poisoning has always been one of the important causes of body injury, but the mechanism of neurotransmitter secretion disorder caused by manganese is not clear at present. Objective To investigate the effects of presynaptic membrane intracellular protein 13-1 (Munc13-1) and synapse fusion protein binding protein 18-1 (Munc18-1) on dopamine secretion dysfunction induced by manganese chloride (MnCl₂) in human neuroblastoma (SH-SY5Y) cells. Methods A SH-SY5Y cell model induced by MnCl₂ was established. Cell viability was measured by MTT assay. Four experimental groups were set up: control group and low-, medium-, and high-dose manganese groups (0, 100, 200, and 400 μmol·L⁻¹ MnCl₂). They were treated with corresponding doses of MnCl₂ for 24 h. The secretion of dopamine was measured by enzyme-linked immunosorbent assay. The mRNA expression of Syntaxin-1 was detected by real-time quantitaive PCR. Total cell proteins were extracted, and the protein expression levels of Munc13-1, Munc18-1, and Syntaxin-1 were detected by Western blotting. The correlations of MnCl₂ exposure and dopamine secretion with the protein expressions of Munc13-1 and Munc18-1 were also analyzed by Pearson correlation. Results Compared with the control group, the cell viability rate decreased gradually with the increase of manganese exposure concentration, and the difference between the medium- and the high-dose manganese groups was statistically significant (P<0.05). The concentration of dopamine in cell culture medium of all manganese exposure groups decreased with the increase of manganese concentration, and compared with the control group and the low-dose manganese group, the medium- and the high-dose manganese groups were statistically significant (P<0.05). The expression of Syntaxin-1 at mRNA or protein level did not change significantly among groups (P>0.05). Compared with the control group, the protein expression of Munc13-1 decreased and that of Munc18-1 increased with the increase of manganese concentration (P<0.05). Compared with the low-dose manganese group, the changes of Munc13-1 protein in the high-dose manganese group and Munc18-1 protein in the medium- and high-dose manganese groups had statistical significance (P<0.05). Compared with the medium-dose manganese group, the protein changes of Munc18-1 in the high-dose manganese group were statistically significant (P<0.05). The correlation analysis showed that MnCl₂ dose was negatively correlated with Munc13-1 protein expression (r=−0.898, P<0.05), and positively correlated with Munc18-1 protein expression (r=0.678, P<0.05). Dopamine secretion was positively correlated with Munc13-1 protein expression (r=0.932, P<0.05), and negatively correlated with Munc18-1 protein expression (r=−0.817, P<0.05). Conclusion The inhibition of dopamine secretion in SH-SY5Y cells induced by manganese exposure is related to up-regulation of Munc18-1 and down-regulation of Munc13-1 expression levels, which may be one of the reasons for nerve injury caused by manganese.

Mutagenesis and screening of hydrogen-producing photosynthetic bacteria,Rhodobacter sp.R7 strain,was investigated by using the combination mutation of ultraviolet ray and LiCl and layer plating methods.A carotenoid mutant named R726 strain was obtained.The plate phenotype properties in carotenoid mutant were different from that of parent strains.Living cells spectra showed that absorption peak of 550 nm was appeared in carotenoid mutant,but not in parent strain.The absorption spectra of extraction of pigment further confirmed the difference of carotenoid composition between the mutant and parent strains.The result of TLC on silica gel plate showed that mutant has a lack of yellow carotenoid composition which occurs in parent strain.H_2 productivity and biomass in carotenoid mutant was higher than that of parent strain.These results revealed that mutant has a modified carotenoid biosynthesis pathway.

Objective To screen for differential expression of genes in bone marrow mesenchymal stem ceils (MSCs) stimulated by electromagnetic fields (EMFs) and to study the mechanism of differentiation of the bone mar- row MSCs induced by EMFs.Methods Mouse bone marrow MSCs were isolated and cultured in vitro.The third- passage cells were stimulated by EMFs,then the total RNA was extracted,purified to cDNA and reversely transcribed to yield a cRNA probe.The cRNA probes from stimulated and control cells were labelled with Cy5 and Cy3,respec- tively.The two samples were hybridized with a mouse oligo microarray,and the hybridization signals were scanned. The reverse transcriptase polymerase chain reaction (RT-PCR) was used to identify the interesting genes:Duspl and Rabl.Results A total of 153 differentially expressed genes were found comparing the stimulated group and the control group.There were 74 up-regulated genes and 79 down-regulated genes in the stimulated group.Semi-quantita- tive RT-PCR confirmed that the Duspl and Rabl mRNA expression levels of the stimulated group were up-regulated. Conclusion The gene expression profiles of the bone marrow MSCs were altered after EMF exposure.The genes are involved in cell proliferation and differentiation,transcription control,metabolism and response to stress.These re- sults provide deeper insight into the mechanism of differentiation of the bone marrow MSCs.

OBJECTIVE To evaluate the use of antibiotics in general hospitals of Ningbo. METHODS Totally 4 391 case history records in April,2004 of 12 hospitals were investigated on the use of antibiotics. RESULTS Medicines incomes accounted for 55.16% in total revenue in a hospital,and antibiotics accounted for 32.66% among medicines incomes.Antibiotics using rate was 62.11% in internal medicine departments.The percentage of antibiotics using without evidence was 23.92%,the combined antibiotics using rate was 49.89%.The average duration for antibiotics using was 10.83 days.Antibiotics using rate in surgery departments before operation was 71.00%,during operation was 20.48%,the combined antibiotics using rate during operation was 14.50%,antibiotics using rate after operation was 96.55%.Antibiotics using for treatment accounted for 62.37% and for prevention was 24.17%. CONCLUSIONS Antibiotics using rate is high in hospitals in Ningbo.Income of medicine is also an important part of total revenue in a hospital.We should pay more attention to management on clinical use of antibiotics.

Objective To investigate the relationship between rapists and related allele genes based on the analysis of 15 short tandem repeats (STRs) loci genetic polymorphism. Methods The method of Genome-wide scan was being used. Buccal swab samples of 129 rapists and 156 random populations were collected and PCR compound amplification was carried out with the aid of AmpFISTR Identifiler system. Then the products were subjected to electrophoresis and gene detection with AB13100 type gene analysis system so as to calculate and compare the alleles of 15 STRs gene frequency in the two groups. Results All the 15 STRs loci allele gene frequency in rapists and random population was found to coincide with Hardy-Weinberg law(P>0. 05). Allele 28 of D21S11 (rapists: 1.55% ,control group:5. 13%) ,allele22 of FGA(rapists:24.03% ,control group:16.99%),allele23 of FGA(rapists: 17.05% ,control group:26.28%) ,allele 10 of TH01(rapists:1.16% ,control group:4.17%) ,allele 8 of TPOX(rapists:55.77% ,control group:63.77%),allele 12 of TPOX(rapists:4.26% ,control group: 1.28%) were different between the two groups (P< 0.05) .while it is no differ significantly in other STRs loci allele gene(P >0.05). Conclusion Allele 28 of D21 S11,allele 22 and 23 of FGA, allele 10 of TH01, allele 8 and 12 of TPOX may be associated with the violent crime of rape. It is suggested that there are existing sensitive or resistance genes about the violent crime of rape in chromosome 2,4,11,21.

Objective To study the effects of 50 Hz magnetic fields on the in vitro proliferation of human osteosareoma cell line MG-63 with different cell densities.Methods Four different magnetic intensities(1 mT, 2 mT,3 roT,4 mT)were used to stimulate the cells,and the experiment was repeated with different cell densities. The method of MTT was employed to evaluate the level of proliferation.Results Fifty Hz magnetic fields signifi- cantly affected the level of proliferation of human osteosareoma cell line MG-63,and the 2 mT intensity exerted the greatest influence on it.The effects of the magnetic field differed with different cell densities.Conclusion The effect of 50 Hz magnetic fields on the in vitro proliferation of human osteosarcoma cell line MG-63 was not only relat- ed to the magnetic intensity,but also the cell density,

Objective To investigate the value of 99Tcm-methoxyisobutylisonitrile (MIBI) scintigraphy in assessing the preoperative chemotherapy response and multidrug resistance of osteosarcoma.Methods From January 2007 to October 2008, 12 patients (female:4, male:8; mean age:16.3 years,range:8-27 years) underwent early (10min) and delayed (120 min) 99Tcm-MIBI scintigraphy before and after preoperative chemotherapy.Seven cases had osteosarcoma at the distal femurs, 2 at the proximal tibias, 2 at the upper end of humerus and 1 at the fibula.The tumor-to-background ratio (T/B) and washout rate (WR) were calculated.Tumor necrosis was classified according to Huvos criterion after limb salvage surgery.Immunohistochemical staining for P-glycoprotein(gp) was examined.Spearman correlation analysis and t-test were performed.Results According to Huvos criterion, 7 patients were classified as good responders with more than 90% of tumor cell necrosis and 5 as poor responders with less than 90% of tumor cell necrosis.R value (ratio of early phase T/B after and before chemotherapy) was significantly lower in good responders than that in poor responders (0.473 ± 0.21 vs 0.998 ± 0.06, t= 5.342, P= 0.000 ).R value was significantly correlated with the degree of tumor cell necrosis ( rs=- 0.87, P= 0.000 ).WR was significantly higher in patients with positive P-gp expression than that in patients with negative P-gp expression ((38.36 ±18.64)% vs (6.40±5.87)%, t= -3.278, P=0.008).There was significant correlation between the WR and P-gp expression (rs = 0.91, P= 0.001 ).Conclusion 99Tcm-MIBI scintigraphy is a feasible non-invasive technique to assess the chemotherapy response and to detect P-gp expression of osteosarcoma.

Objective To explore the temporal and spatial expression pattern of tenascin-c(tnc) and tenascin-w(tnw) in zebrafish early development,to further explore the role of tenacsin in zebrafish embryo development,and the association between them.Methods Zebrafish embryos at 2 hours post fertilization(hpf),4 hpt,8 hpt,10 hpf,24 hpt,48 hpr,72 hpf and 7 days post fertilization(dpf) were collected to extract RNA for reverse transcription-polymerase chain reaction(RT-PCR) and fix the embryos at different stages for in situ hybridization.Temporal and spatial expression pattern of tnc and tnw on different stages of zebrafish early development was observed.Results tnc and tnw all expressed in zebrafish from 24 hpf to 7 dpf,but did not expressed from 2 hpf to 10 hpf.Tnc expressed at pharyngeal arch,notochord,somite in 24 hpf,then weakly expressed at somite,but highly expressed at otic vesicle,pectoral fin and hindbrain in 48 hpf,and tnc was expressed at hindbrain,pharyngeal and notochord and disappeared at somite and pectoral.tnw expressed at hindbrain,midbrain and otic vesicle in 24 hpf,expressed at somite,notochord,hindbrain,otic vesicle and pharyngeal in 48 hpf.In 72 hpf,tnw expressed weakly at somite and notochord.Conclusions Zebrafish tnc and tnw have special temporal and spatial expression pattern,and share partial overlapping expression pattern.

Objective To explore the expression pattern of tenascin-c(tnc)gene in zebrafish embryo abnormal development which was induced by ethanol,and to further understand the function of tnc gene in embryo develepment.Methods Zebrafish were treated with ethanol at different concentration from 100 to 500 mmol/L,and embryos at 24 and 48 hours were collected and fixed,then tnc expression pattern was observed by in situ hybridization and reverse transcriptase-polymerase chain reaction(RT-PCR).Results The result of RT-PCR showed that ethanol at 100 and 200 mmol/L could increase the expression of tnc,while the result of in situ hybridization showed that,while ethanol at 300 mmol/L and above decrease the expression of tnc in presumptive position at 24 hours,and ethanol at 100 mmol/L and above caused increase expression of tnc in zebrafish heart.Conclusions tnc is increased when treated with 100 and 200 mmol/L ethanol and is presented in the abnormal development of hearts of zebrafish,which can promote the normal development of embryos in some degrees.The expression pattern of tnc in pathologic state is highly conserved in all vertebrate,and in adult and embryos as well.