OBJECTIVETo construct microRNA-223 overexpression and suppression lentivirus vectors and determine their effects after infecting oral squamous cell carcinoma (OSCC) cell line.

METHODSLentivirus vectors GV229 and GV232 were cut by the restriction sites of Age I and EcoR I and connected to the target gene, which contained mature microRNA-223 and microRNA-223 oligonucleotide. Real-time polymerase chain reaction (PCR) method was used to detect the microRNA-223 expression level after infecting the recombinant lentivirus vector into the OSCC cell line.

RESULTSThe successful construction of microRNA-223 recombinant lentivirus vectors was confirmed by the PCR method and DNA sequencing. HN-30 cell infected with microRNA-223 overexpression vector showed a significant increased in microRNA-223 expression, whereas HN-30 cell infected with microRNA-223 inhibitor vector suppressed microRNA-223 expression.

CONCLUSIONThe microRNA-223 overexpression and suppression lentivirus vectors are successfully constructed. These vectors could alter the expression level of microRNA-223 in OSCC cell line significantly, and provide a stable cell line for functional studies in the future.

Base Sequence ; Cell Line, Tumor ; Genetic Vectors ; Humans ; Lentivirus ; MicroRNAs ; metabolism ; Real-Time Polymerase Chain Reaction

PurposeSmall breast mass (diameter≤1 cm) is prone to misdiagnosis in clinic. This paper aims to evaluate a combined application of breast imaging reporting and data system (BI-RADS) and ultrasound elastography (UE) on small breast mass (diameter≤1 cm).Materials and MethodsA retrospective analysis was carried out on 231 patients with a total of 258 small masses (the maximal diameter ≤1 cm). Ultrasound BI-RADS was used for classiifcation while UE was used to adjust the results. The results were further compared with those of postoperative pathology. The curve of ROC was employed to evaluate the combined use on small breast mass.ResultsAmong the 258 small masses, 178 (69.0%) were benign masses and the rest 80 (31.0%) were malignant. The small masses which were evaluated as BI-RADS grade 3, 4 and 5 before the operation had positive prediction value for malignant masses of 10.3% (17/165), 60.5% (46/76) and 100.0% (17/17), respectively. After adjustment with UE, the values changed to 5.3%(9/169), 75.0% (54/72) and 100.0% (17/17), respectively. After adjustment with the combination method, the area under ROC curve in BI-RADS classification was 0.904, which was signiifcantly higher than that (0.827) before the adjustment (Z=2.83,P<0.05). ConclusionFor small breast mass (diameter≤1 cm), mass of BI-RADS grade 3 has higher positive prediction value. But after adjustment with UE, the positive prediction value of mass of BI-RADS grade 3 tends to be lower, whilst that of mass of BI-RADS grade 4 increases, thus promoting the efficiency of ultrasound BI-RADS classification for small breast mass and contributing to the identiifcation of benign and malignant small breast masses.

Objective To investigate the significance of metastasis associated lung adenocarcinoma transcript 1 ( MALAT1 ),cyclooxygerase-2 ( COX-2 ),beta catenin ( β-catenin )、matrix metalloproteinase (MMP)-3 and MMP-9 in the occurrence and development of colorectal carcinoma.Methods Real-time PCR was used to detect MALAT1,COX-2,β-catenin,MMP-3 and MMP-9 rnRNA expression in samples from 30 fresh colorectal carcinomas and 30 corresponding adjacent tissues.And the correlation analysis of the gender and age of patients,CEA,immune cellular factors ( CD4 and CD8 ),clinical stages,and the degree of differentiation was undertaken.Results The expression levels of MALAT1,COX-2,β-catenin and MMP-9 were significantly different between colorectal carcinoma tissues and adjacent colorectal tissues (P ＜ 0.05 ).MMP-3 showed no significant difference (P ＞ 0.05).MALAT1,COX-2,β-catenin and MMP-9 expression levels showed an average 2.22-fold,1.86-fold,2.16-fold,0.58-fold ( P ＜ 0.01 ) increase in colorectal carcinoma tissues when compared with adjacent colorectal tissues respectively.There were negative correlation between MALAT1 and β-cateuin ( colorectal carcinoma tissues vs adjacent colorectal tissues) ( r =- 0.346,P =0.030).While there were positive correlation between MMP-9 and β-catenin ( colorectal carcinoma tissues vs adjacent colorectal tissues) ( r =0.312,P =0.047 ).There were significant difference between male patients and female patients in terms of COX-2 and MMP-9 (colorectal carcinoma tissues vs adjacent colorectal tissues) (P =0.047; P =0.018).There were significant difference between patients with tumor marker CEA increase and patients without CEA increase in terms of COX-2 ( colorectal carcinoma tissues vs adjacent colorectal tissues) ( P =0.021 ).Conclusion MALAT1,COX-2,MMP-9 and β-catenin have significance in the occurrence and development of colorectal carcinoma,while MMP-3 has no significant reference value. The negative correlation between MAL(A)T-1 and β-catenin and the positive correlation between MMP-9 and β-catenin might show some interaction relationship in the development of colorectal carcinoma.The expression differences of COX-2 and MMP-9 (colorectal carcinoma tissues vs adjacent colorectal tissues) in male and female patients suggest that above two genes may affect the occurrence ratio of colorectal carcinoma.Detecting of COX-2 maybe helpful to the tumor marker CEA during the diagnosis of colorectal carcinoma.

Objective To identify the gene mutation of G protein?-subunit (Gsct) in multiple affected tissues of a patient with McCune-Albright syndrome.Methods The peripheral blood,bone tissue,lesion skin and pleura samples of the patient were collected.Genomic DNA was isolated from these samples,and PCR and direct sequencing were performed.Results The peripheral blood and bone tissue of the patient showed a mutation R201C in Gs?gene.No mutation was detected in the skin and pleura samples of the patient.Conclusion The gene diagnosis confirms that the patient has a classical R201C mutation in Gs?gene and multiple tissues are affected.The mutation occurs early in embryogenesis and clinical features can be polymorphic.

Sulfur fumigation (SF) is a universal phenomenon in primary processing of Traditional Chinese Medicine (TCM) in modern times. In the process, fumigation, sulfur or both of them act on the TCMs. Some active components of TCMs change quantitatively or qualitatively during the processing. At the same time, the sulfur dioxide and heavy metal would remain and cause a serious influence on quality and future development of TCM. This article reviews the chemical compositions change after SF to study the change law and their influence on quality. This article provide references for SF in TCMs' processing for a better and safer quality.
Drug Contamination ; Fumigation ; methods ; Medicine, Chinese Traditional ; methods ; Quality Control ; Sulfur ; chemistry ; Technology, Pharmaceutical ; methods

Vascular endothelial growth factor 121 (VEGF121) was expressed as inclusion bodies by recombinant Escherichia coli. High concentrations of both biomass (46 g dry cell/L) and VEGF121 inclusion bodies (4.5 g/L) were obtained by applying a high-cell-density culture. After the inclusion bodies were washed and dissolved, VEGF121 was refolded at 0.2 mg/ml by ultrafiltration in refolding buffer with a yield of 81%. Renatured VEGF121 was purified by anion chromatography and Sephacry S-100 chromatography with purity higher than 95% and fnal purification yield of 31%. The purified VEGF121 could stimulate the proliferation of human umbilical vein endothelial cells as demonstrated by a biological activity assay.

Background Leber hereditary optic neuropathy (LHON)is a common inherited eye disease,which generally affects young adults with bilateral loss of central vision.Mutation frequency of Leber hereditary has not been fully clarified. Objective This study was to investigate the mutation frequency of mitochondrial NDI gene associated with LHON in Chinese population. Methods The proposal of the study was approved by Ethic Committee of Wenzhou Medical College.Written informed consent was obtained from each subject initial of this trial.Eight hundred and ninety-four LHON patients and 134 normal subjects were collected.Genomic DNA was extracted from peripheral blood leukocytes of the all participants.Polymerase chain reaction (PCR) was used to amplify and sequence analysis of the mitochondrial ND1 gene was performed and aligned with revised Cambridge Reference Sequence(rCRS) of mitochondrial DNA.Then mutated gene frequency was screened and analyzed. Results Mutational analysis of mitochondrial ND1 gene in 894 LHON patients revealed the presence of G3316A,T3394C,G3460A,C3497T,G3635A,G3733A,and T4216C.11.19％ LHON patients (100/894 ) were found to be associated with the gene mutations mentioned above,and 3.24％ patients (29/894) showed the co-occurrence of three primary mutations.Mutation frequencies in LHON patients were 2.57％,2.23％,1.45％,3.80％,0.67％,0.11％,0.34％,respectively,and G3316A,T3394C,C3497T and T4216C also were detected in 134 normal controls with the mutation frequencies of 4.48％,2.99％,4.48％ and 1.49％,respectively.Mutation frequency analysis showed an insignificant difference in the mutations of G3316A,T3394C,C3497T and T4216C between LHON patients and normal controls (x2 =0.926,P=0.336;x2 =0.052,P=0.820; x2 =0.142,P=0.707;P=0.129).G3376A,G3496T,G3700A,A4136G,T4160C and C4171A were absent in Chinese LHON patients. Conclusions Mitoehondrial ND1 gene in LHON is a mutational hotspot in Chinese population,11.19％ (100/894)associated with LHON was caused by ND1 gene mutation.G3635A,G3733A may be rare pathological mutation in Chinese population.However,G3316A,T3394C,C3497T and T4216C are insufficient to produce the clinical phenotype,but they may play a synergic role for penetrance and phenotypic manifestation in LHON.

Fifteen known compounds were isolated from Swertia delavayi by silica gel, Sephadex LH-20 and Rp-18 column chromatographies. Based on extensive spectroscopic analysis (MS, 1H, 13C-NMR), their structures were identified aserythrocentaurin (1), erythrocentaurindimethylacetal (2), sweroside (3), swertiamarin (4), gentiopicroside (5), swertiakoside A (6), 2'-O-acetylswertiamarin (7), 4'-O-[(Z) -coumaroyl] swertiamarin (8), 1,5,8-trihydroxy-3-methoxyxanthone (9), 8-O-β-D-glucopyranosyl-1-hydroxy-2,3, 5-trimethoxyxanthone (10), 8-O-[β-D-xyl- opyranosyl-(1 --> 6)-β-D-glucopyranosyl]-7,8-dihydroxy-3-methoxyxanthone (11), isovitexin (12), β-sitosterol (13), daucosterol (14), and oleanolic acid (15). Among them, ten ones (14, 7-11, 13) were obtained from S. delavayi for the first time. The isolates were evaluated for their anti-HBV activities in HepG 2. 2. 15 cell line in vitro. The results showed that compound 1, 2, 6, 7, 9 and 12 exhibited significant inhibitory activity on HBV DNA replication with IC50 values from 0.05 to 1.46 mmol x L(-1).
Antiviral Agents ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Hepatitis B virus ; drug effects ; genetics ; Magnetic Resonance Imaging ; Molecular Structure ; Spectrometry, Mass, Electrospray Ionization ; Swertia ; chemistry

OBJECTIVETo study the genetic polymorphism of 15 short tandem repeat (STR) (D2S1338ì D3S1358ì D5S818ì D7S820, D8S1179, D13S317, D16S539, D18S51, D19S433, D21S11, CSF1PO, TPOX, TH01, vWA, FGA) in Maonan minority of Guangxi province.

METHODSThe allele frequencies and the genotype of 15 STR loci were analyzed in 143 unrelated individuals in Maonan minority of Guangxi by PCR-STR and genescan.

RESULTSThere were 130 STR alleles and 390 genotypes in the 15 STR of Maonan minority, with allele frequencies ranging from 0.0035 to 0.5385. The average heterozygosity was 0.7697, the discrimination power was higher than 0.8 except for that of TPOX, the accumulative discrimination power was more than 0.999999999, and the probability of paternity exclusion was more than 0.99999918.

CONCLUSIONThe 15 STR loci of Maonan minority in Guangxi possesses the characteristics of high genetic diversity, except for the TPOX locus. They can be employed in minority genetics investigation, individual and paternity test in forensic medicine.

Adult ; Aged ; Aged, 80 and over ; Alleles ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Microsatellite Repeats ; genetics ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics