The identiifcation of body lfuid and their tissue resource is an important part of forensic medicine research. Conventional methods have imperfections like high false positive rate and may destroy biomaterial, which should be replaced by a new conifrmatory test. Highly-differentiated cells express speciifc mRNA molecules that can be used for body lfuid identiifcation. These mRNA markers can identify body lfuids like peripheral blood, menstrual blood, semen, saliva, vaginal secretion, skin-contact stains. This method have favorable sensitivity, speciifcity and can be detected after years, made it a ideal way to identify current body lfuids in the future. This review focus on the application of mRNA in body lfuid identiifcation and tissue resource.

Objective To explore the expression of tankyrase (TNKS) and its relationship with WNT/β-catenin signal?ing pathway in lung acinar adenocarcinoma. Methods Seventy-two samples of single subtype alveolar like lung adenocarci?noma (lung adenocarcinoma group) and 67 specimens of normal lung tissue adjacent to carcinoma (adjacent to carcinoma group) were collected. Immunohistochemical method was used to detect expressions of TNKS, beta-catenin (β-catenin) and c-myc protein. The correlation of each protein expression in lung adenocarcinoma tissues was analyzed. The differential ex?pression of TNKS was detected by Western blot assay in two groups. Results Tankyrase protein was mainly expressed in cy?toplasm. The expression ofβ-catenin protein was mainly in cytoplasm and nuclear of lung adenocarcinoma. The expression ofβ-catenin was mainly in cytoplasm, and a small amount was in nuclear of the adjacent group. The c-myc protein was ex?pressed mainly in the nucleus. The positive expression rates of TNKS,β-catenin and c-myc protein were significantly high?er in lung adenocarcinoma group than those of adjacent to carcinoma group (P<0.05). The expression ofβ-catenin in cyto?plasm and nucleus was positively correlated with the expression of TNKS and c-myc (P<0.05). Western blot analysis showed that the relative expression level of TNKS was significantly higher in lung adenocarcinoma group than that of adja?cent to carcinoma group (0.497 ± 0.021 vs. 0.237 ± 0.015, t=13.00, P<0.01). Conclusion Abnormally high expression of TNKS in lung adenocarcinoma may promote the occurrence of lung cancer by regulating the WNT signaling pathways. Inhib?iting TNKS expression may become a new target to treat lung adenocarcinoma.

In order to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix using ITS2 barcodes, genomic DNA from sixty samples was extracted and the ITS2 (internal transcribed spacer) regions were amplified and sequenced. The genetic distances were computed using MEGA 5.0 in accordance with the kimura 2-parameter (K2P) model and the neighbor-joining (NJ) phylogenetic tree was constructed. The results indicated that for Aucklandiae Radix (Aucklandia lappa), Vladimiriae Radix (Vladimiria souliei and V. souliei var. cinerea), Inulae Radix (Inula helenium), Aristolochiae Radix (Aristolochia debilis) and Kadsurae Radix (Kadsura longipedunculata), the intra-specific variation was smaller than inter-specific one. There are 162 variable sites among 272 bp after alignment of all ITS2 sequence haplotypes. For each species, the intra-specific genetic distances were also smaller than inter-specific one. Furthermore, the NJ tree strongly supported that Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix can be differentiated. At the same time, V. souliei (Dolomiaea souliei) and V. souliei var. cinerea( D. souliei var. cinerea) belonging to Vladimiriae Radix were clearly identified. In conclusion, ITS2 barcode could be used to identify Aucklandiae Radix, Vladimiriae Radix, Inulae Radix, Aristolochiae Radix and Kadsurae Radix. Our study may provide a scientific foundation for clinical safe use of the traditional Chinese medicines.
Aristolochia ; classification ; genetics ; Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plants, Medicinal ; classification ; genetics ; Quality Control

To investigate the expression patterns of peroxisome proliferator activated receptor2 (PPARgamma2) gene in the differentiation of mouse embryonic stem (ES) cells into adipocytes, mouse ES cells were transfected with the vector of pPPARgamma2-promoter-luciferase, and PPARgamma2 expressions were analyzed by detecting luciferase activities and by detecting the protein expressions using western blotting. The results showed that the gene PPARgamma2 did not express in undifferentiated mouse ES cells and in embryoid bodies (EBs) within the first 2 days of differentiation induction after EB formation, and began to express from the third day of differentiation induction after EB formation to the finish of the differentiation. The gene's expression in differentiated adipocytes was much stronger than that in differentiating preadipocytes. In Conclusion our results reported for the first time the five-step expression patterns of the gene PPARgamma2 during the whole differentiation procedures from mouse ES cells into adipocytes via preadipocytes, and supported the previous studies that PPARgamma2 is a fat-specific gene that expresses only in developed and developing adipose tissues.
Adipocytes ; cytology ; Animals ; Cell Differentiation ; genetics ; Cells, Cultured ; Electroporation ; Embryonic Stem Cells ; cytology ; metabolism ; Mice ; PPAR gamma ; biosynthesis ; genetics ; Promoter Regions, Genetic ; genetics ; Transfection

A cell model is desired for adipocyte differentiation investigation and for high-throughput screening of anti-obesity and anti-diabetes molecules from chemical resources due to the world wide epidemic of obesity and diabetes. In order to establish such a cell model, a plasmid of pPPARgamma2-promoter-EGFP was constructed by inserting a 660bp sequence of mouse PPARgamma2 promoter into the Ase I and Kpn I sites of pEGFP-N3 and transferred into 3T3-L1 preadipocyte cells. The cells were induced to differentiate and the expression of PPARgamma2 was detected by the microscopic observation of EGFP and by RT-PCR assays. The results showed that the EGFP gene expression patterns were similar to that of pPPARgamma2's, which indicated that the EGFP gene was transferred into the mouse 3T3-L1 preadipocyte cells, and its expression was under the control of pPPARgamma2 promoter. RT-PCR assays showed that the EGFP expression authentically represented the stable expression of PPARgamma2. In conclusion, a preadipocyte cell line expressing EGFP under the control of the promoter of adipocyte-specific expression gene PPARgamma2 was generated. The cell line provides a powerful approach for the research of adipocyte differentiation and for the high-throughput screening of anti-obesity and anti-diabetes chemicals.
3T3-L1 Cells ; Adipocytes ; cytology ; Animals ; Cell Differentiation ; Green Fluorescent Proteins ; genetics ; Mice ; PPAR gamma ; genetics ; Promoter Regions, Genetic ; Stem Cells ; cytology

Objective To evaluate the adaption of the condyle position of patients with extensive tooth wear during occlusal rehabilitation,and the correlation between increased vertical dimensions and the changes of joint spaces.Methods Twenty-seven patients(five from Beifang hospital,others from Peking University School and HospitM of Stomatology)with extensive tooth wear were selected and received occlusal 1),1 month following delivery of temporary restoration(stage 2),and 1 month following delivery of permanent restoration(stage 3).Results The superior,anterior and posterior joint spaces were(3.24±0.16),(2.06±0.11),(1.89±0.13)mm at stage 1;(3.61±0.15),(1.94±0.10),(2.52±0.11)mm at stage 2;(3.49±0.19),(1.93±0.10),(2.40±0.13)mm at stage 3.The posterior joint spaces at stage 2 and stage 3 were significantly larger than that at stage 1(P<0.01).The superior spaces at stage 2 were significantly larger than that at stage 1(P<0.05).No correlations between the increased vertical dimensions and the changes of joint spaces were found in the three stages(P>0.05).Conclusions The condyle positions in the patients with extensive tooth wear changed after occlusal rehabilitation.

Objective To compare diversity and diagnostic concordance of Subjective Global Assessment (SGA) with Nutritional Risk Screening 2002 (NRS2002) in evaluating nutritional status among gastrointestinal cancer patients. Methods Two hundred and twenty-three patients with gastrointestinal cancer were conveniently selected and evaluated by SGA and NRS within 48h of admission from April to October 2009 using cross-sectional study. Diversity and diagnostic concordance between SGA and NRS2002 were analyzed by McNemar paired chi- square test and Kappa test. Results All patients completed both NRS2002 and SGA. The prevalence of nutritional risk or undernutrition evaluated by NRS2002 or SGA was 61% versus 32. 3%. There was a significant difference between NRS2002 and SGA after conducting McNemar paired chi-square test (P <0. 01) but diagnostic concordance of SGA and NRS2002 was poor (Kappa = 0.368, P < 0. 01). Conclusions Both NRS2002 and SGA were feasible in evaluating nutritional status in gastrointestinal cancer patients, but there was deference between NRS2002 and SGA. NRS2002 had higher sensitivity to detect early under nutrition and was more suitable in prevention under nutrition than SGA in gastrointestinal cancer patients.

·AIM: To evaluate the efficacy of anti - vascular endothelial growth factor (VEGF) and photodynamic therapy (PDT) on pathological myopia (PM) combined with choroidal neovascularization (CNV). ·METHODS: Forty-three patients (45 eyes) diagnosed by fundus fluorescein angiography (FFA), indocyanine green angiography ( ICGA ) and optical coherence tomography (OCT) with PM combined with macular CNV were recruited in this study. The patients were randomly divided into two groups for different treatments, intravitreal injection with Ranibizumab (20 patients, 22 eyes) and PDT(23 patients,23 eyes). After treatment,all patients had been followed up monthly for 12mo. The further treatments were operated according to referral situations. The best corrected visual acuity (BCVA) was recorded with the ETDRS chart and the mean defect(MD) of the center 10° visual field was measured. At the last follow-up,the therapy efficacy was determined by ETDRS numbers and MD and analyzed. ·RESULTS: Before treatment, there was no significant difference on the baseline in ETDRS and MD between ranibizumab group and PDT group (P>0.05). After 12mo treatment, the ETDRS number in ranibizumab group (39.23±20.06) significantly increased (by 5.88 ± 9.03, P<0.05), but the PDT group (37. 38 ± 16. 95) was not significantly improved(by 0.33±6.94,P>0.05). The MD in ranibizumab group decreased significantly (P<0.05), and no significant change was found in PDT group(P>0.05). · CONCLUSION: In the treatment of macular CNV complicated by the PM, ranibizumab injection can improve visual function better than PDT.

·AIM: To evaluate the efficacy of anti - vascular endothelial growth factor ( VEGF ) and photodynamic therapy ( PDT) on pathological myopia ( PM) combined with choroidal neovascularization ( CNV ) using ETDRS chart and multifocal electroretinogram (mfERG). ·METHODS: Forty-three patients ( 45 eyes) diagnosed by fundus fluorescein angiography ( FFA ), indocyanine green angiograph ( ICGA ) and optical coherence tomography (OCT) with PM combined with macular CNV were recruited in this study. The patients were randomly divided into two groups for different treatments, intravitreal injection with Ranibizumab ( 20 patients, 22 eyes) and PDT (23 patients, 23 eyes). After treatment, all patients had been followed up monthly for 12mo. The further treatments were operated according to referral situations. The best corrected visual acuity ( BCVA) was recorded with the ETDRS chart and mfERG. At the last follow-up, the therapy efficacy was determined by ETDRS numbers and mfERG and analyzed. ·RESULTS: Before treatment, there was no significant difference on the baseline in ETDRS and mfERG latency of N1 wave, latency and the density values of P1 wave between ranibizumab group and PDT group. After 12mo treatment, the ETDRS number in ranibizumab group (39. 23± 20. 06) significantly increased to the baseline by 5. 88±9. 03(P<0. 05), and in PDT group (37. 38±16. 95) was not significantly improved by 0. 33±6. 94(P>0. 05). There was no significant difference in latency of N1 wave, latency and the density values of P1 wave from treatment response of mfERG in the two groups (P>0. 05). · CONCLUSION: In the treatment of macular CNV complicated by the PM, ranibizumab injection can improve visual function better than PDT, while similar on macular.

OBJECTIVETo evaluate the clinical performance of the two configurations of gingival margin preparation of tooth.

METHODSThe cases in this study were divided into two groups according to different tooth defects. Each group consisted of 40 cases. One group's gingival margin configuration was 90 degree shoulder, the other was under-gingival non-shoulder. The clinical performance of these restorations was followed up for 1 year and 2 years. The evaluators examined the restorations for plaque index, gingival index, marginal discolor and marginal fit.

RESULTSThere was no significant difference in the evaluators between the two groups.

CONCLUSIONThe under-gingival non-shoulder margin configuration of porcelain fused to metal should be used for clinical application compared with the 90 degree shoulder one in certain circumstance.