Objective To prepare the 99Tcm-survivin mRNA antisense peptide nucleic acid (PNA)and investigate its value as a gene imaging agent in tumor bearing mice and early diagnosis in tumor.Methods Survivin mRNA antisense PNA and mismatch PNA were synthesized.Four amino acids (Gly- (D)Ala-Gly-Gly) and Aba (4-aminobutyric acid) were linked to the 5' end of PNA.Gly- (D)Ala-Gly-Gly served as a chelating moiety for strong chelation of 99Tcm and Aba acted as a spacer to minimize the steric hindrance.PNAs were labeled with 99Tcm by the ligand-exchange method.The labeling efficiency and radiochemical purity were measured by HPLC and ITLC methods.There were five BALB/c nude mice bearing human lung carcinoma ( A549 ) in each of antisense PNA and mismatch PNA groups.Gene imaging of 99Tcm-survivin mRNA antisense and mismatch PNAs were performed at 1,2 and 4 h post the injection,respectively,and the T/NT ratio was measured by the method of ROI.The statistical comparisons of average values were performed with the two-group t-test for independent sample by SPSS 13.0.Results The product kept stable in vitro.The labeling efficiency of 99Tcm-survivin mRNA antisense PNA was (95.48 ±1.92)％ and more than 85％ after the incubation for24 h in serum.The radiochemical purity was ＞ 95％.The labeling efficiency of mismatch PNA was similar to the antisense PNA.99Tcm-survivin mRNA antisense PNA was especially uptaken by tumor lesion,and its accumulation reached the top at 4 h post the injection.T/NT ratios at 1,2,and 4 h were 2.70 ± 0.28,3.44 ± 0.35,4.21 ± 0.63,respectively.In the comparison,the T/NT ratio of 99Tcm-survivin mRNA mismatch PNA at 4 h (3.12 ±0.50) was significantly lower (t =2.918,P =0.019).Conclusions 99Tcm-survivin mRNA antisense PNA has high labeling efficiency,good stability and no need of purification.Its characteristic of especial uptake by tumor lesion provides the potential value in early diagnosis of tumor.

OBJECTIVETo observe the interventional effect of electroacupuncture combined with catgut implantation at acupoints for treatment of simple obesity of heart and spleen deficiency type.

METHODSSixty five cases were randomly divided into an observation group (33 cases) and a control group (32 cases). The observation group was treated with electroacupuncture combined with catgut implantation at acupoint therapy, the electroacupuncture was applied at Zhongwan (CV 12), Xiawan (CV 10), Guanyuan (CV 4), Tianshu (ST 25), ect. and catgut implantation was given at Zhongwan (CV 12), Tianshu (ST 25), Qihai (CV 6), etc. The control group was treated with electroacupuncture only. The body weight, body mass index (BMI), waistline, waist hip ratio (WHR), Pittsburgh sleep quality index (PSQI), Hamilton anxiety scale (HAMA) and Hamilton depression scale (HAMD, 17 items) were evaluated before and after treatment, and these were also compared with those of 35 nomal cases.

RESULTSThe total effective rate of 93.9% in the observation group was higher than that of 84.4% in the control group (P < 0.05); the body weight, BMI, waistline, WHR, PSQI, HAMD and HAMA of simple obesity cases were obviously higher than those of normal cases (all P < 0.05). The scores of above indexes were all obviously decreased in both groups after treatment (all P < 0.05), and the improvement was more significant in observation group (P < 0.05).

CONCLUSIONThe sleep quality reduction and mental and psychology disorder exist in simple obesity patients, and electroacupuncture combined with catgut implantation at acupoints can reduce weight effectively, and at the same time improve the sleep quality and regulate psychological state.

Acupuncture Points ; Adult ; Catgut ; Electroacupuncture ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Obesity ; physiopathology ; psychology ; surgery ; therapy ; Prostheses and Implants ; Prosthesis Implantation ; Spleen ; physiopathology ; Treatment Outcome ; Young Adult

ObjectiveTo investigate the value of 99Tcm labeled survivin mRNA antisense peptide nucleic acid (PNA) as an imaging agent in the specific diagnosis for carcinoma.MethodsSurvivin mRNA antisense PNA was labeled directly with 99Tcm by the ligand-exchange method.Twenty nude mice with lung carcinoma A549 xenografts were randomly divided into 4 groups.Three groups were used for biodistribution study and one group was used for imaging study.Other twenty mice infected by staphylococcus aureus underwent the same procedure.The biodistribution and imaging of 99Tcm-survivin mRNA antisense PNA was studied at 1,2 and 4 h respectively after the intravenous injection in nude mice bearing lung carcinoma A549 xenografts or inflammation models.SPSS 13.0 was used in the study and all data were analyzed by t test.ResultsBiodistribution results showed that the highest radioactivity was found in the liver,and then in the kidney.Four hours after the administration of the imaging agent,the radioactivity ratios of target-tonon target (T/NT,tumor or inflamumatory lesions to the contralateral regions) in tumor model group were significantly higher than those in inflammation model group ( 3.69 ± 1.13 vs 2.03 ± 0.47,t =3.01,P =0.02 ).Tumors were clearly visible in the tumor model groups at 0.5 h and still clearly seen at 4 h after the injection of antisense PNA.On the contrary,inflammatory lesions could not be seen clearly.Conclusion 99Tcm labeled survivin mRNA antisense PNA can be used to distinguish tumor from inflammation and it may provide a new feasible method for specific tumor diagnosis.

OBJECTIVETo observe the effects of paired box gene 8 (Pax-8) silencing by RNA interference on mitochondrial function and cardiomyocytes apoptosis.

METHODSThe cultured H9C2 (2-1) myocytes were divided into 3 groups: short interference RNA targeting Pax-8 (Pax-8 siRNA) group, non-specific siRNA group as the negative control (NC siRNA), and blank control group (BC siRNA). Fluorescence spectrophotometry was used to detect the activity of caspase-3. RT-PCR was performed to detect mRNA expression of Bcl2 and Bax. The protein expression of Bcl2, Bax and cytoplasm of Cytochrome was examined by Western blot. Changes of ΔΨm were detected by flow cytometry.ΔΨm with JC-1 monomer/polymer ratio was calculated for measuring mitochondrial depolarization proportion.

RESULTSCompared to NC siRNA and BC siRNA group (0.075 ± 0.021, 0.072 ± 0.019), the activity of caspase-3 in Pax-8 siRNA group (0.167 ± 0.012) was significantly increased (P < 0.05); Bcl2 mRNA and protein expression in Pax-8 siRNA group (0.61 ± 0.06, 0.94 ± 0.11) were significantly downregulated compared with NC siRNA group (0.90 ± 0.070, 1.39 ± 0.15) and BC siRNA group (0.94 ± 0.087, 1.49 ± 0.20) (P < 0.05); Bax mRNA and protein expression in Pax-8 siRNA group (1.05 ± 0.10, 1.25 ± 0.12) were markedly upregulated compared with NC siRNA group (0.72 ± 0.03, 0.99 ± 0.12) and BC siRNA group (0.64 ± 0.03, 0.92 ± 0.06), P < 0.05; cytosolic cytochrome expression in Pax-8 siRNA group (0.75 ± 0.14) was significantly upregulated compared with NC siRNA group (0.51 ± 0.06) and BC siRNA group (0.48 ± 0.07) (P < 0.05); JC-1 monomer/polymer ratio in Pax-8 siRNA group (0.163 ± 0.011) was significantly increased compared with NC siRNA group (0.092 ± 0.015) and BC siRNA group (0.072 ± 0.025) (P < 0.05) indicating mitochondrial membrane potential was significantly reduced in Pax-8 siRNA group. Above parameters were similar between NC siRNA group and BC siRNA group (P > 0.05).

CONCLUSIONInhibiting Pax-8 results in enhanced cardiomyocytes apoptosis through the mitochondrial pathway.

Animals ; Apoptosis ; Cells, Cultured ; Mitochondria, Heart ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; PAX8 Transcription Factor ; Paired Box Transcription Factors ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; Rats ; Transfection

OBJECTIVETo investigate the role of human angiopoietin-1 (Ang1) in tumorigenesis and angiogenesis of human gastric cancer cell line SGC7901 in nude mice.

METHODSRecombinant human Ang1 sense or antisense eukaryotic expression vectors were constructed, and transfected by lipofectin into human gastric cancer line SGC7901. Stable transfectants were obtained respectively, namely 7Ang1+ for sense, 7Ang1- for antisense, and 7901P for empty vector transfected cells. Semiquantitative PCR and Western blot were employed to testify the transfection efficiency. Cell growth curve and cell cycle were observed by MTT assays or flow cytometry. In in vivo study, growth of SGC7901 xeno-transplant was observed in BALB/c nude mice. Microvessel density (MVD) was analyzed by immunohistochemistry for Factor VIII staining.

RESULTSStably transfected cell lines were established and decreased expression of Ang1 protein and mRNA in the antisense transfected SGC7901 cells was achieved. Tumorigenesis of 7Ang1- cells on day 30 days was significantly inhibited with decreased MVD as compared to that in 7901P and 7Ang1+ cells (P < 0.01).

CONCLUSIONAngiopoietin-1 plays an important role in tumorigenesis and angiogenesis of gastric cancer which can be partially abrogated by antisense technique.

Angiopoietin-1 ; biosynthesis ; genetics ; Animals ; Cell Line, Tumor ; DNA, Antisense ; genetics ; Genetic Vectors ; Humans ; Mice ; Mice, Nude ; Microcirculation ; pathology ; Neoplasm Transplantation ; Neovascularization, Pathologic ; RNA, Messenger ; biosynthesis ; genetics ; Stomach Neoplasms ; blood supply ; metabolism ; pathology ; Transfection

OBJECTIVETo use COI gene on the Mauremys reevesii and its adulterants by molecular identification. Search a rapid, accurate method of identification of Teseudinis Carapax et Planstrum and its adulterants.

METHODWe collected 8 species of the authentic and adulterants of teseudinis carapax et planstrum in a nationwide then, extracted DNA, got the COI sequences. Use ContigExpress, Dnaman, Edit Sequence and Mega 5 to analyze the variable site and construct the N-J tree.

RESULTCompare with the authentic Teseudinis Carapax et Planstrum, the adulterant exist lots of variable site. The N-J tree Indicates that the same genus belong together and each species belong to relatively independent branch.

CONCLUSIONBased on the COI gene, the technology of DNA bar code can be a excellent identification of Teseudinis Carapax et Planstrum and its adulterants.

Animals ; Base Sequence ; DNA Barcoding, Taxonomic ; Electron Transport Complex IV ; genetics ; Medicine, Chinese Traditional ; standards ; Molecular Sequence Data ; Phylogeny ; Quality Control ; Reptilian Proteins ; genetics ; Sequence Analysis, DNA ; Turtles ; classification ; genetics

OBJECTIVETo observe the clinical effect of dog-day acupuncture and tortoise-shell moxibustion combined with pelvic floor muscle exercises for treatment of female stress urinary incontinence.

METHODSSeventy one cases were randomly divided into two groups. Thirty six cases in the observation group were treated with acupuncture on Zhongji (CV 3), Zigong (EX-CA 1), Chize (LU 5) etc. and tortoise-shell moxibustion on Shenque (CV 8) combined with pelvic floor muscle exercises; while thirty five cases in the control group were treated with only pelvic floor muscle exercises. The scores of the International Consultation Committee on Incontinence Questionnaire Short Form (ICI-Q-SF) and the Medical Outcomes Survey Short Form-36 (SF-36) were evaluated before and after treatment, and the scores of SF-36 were also compared with 35 cases in normal group.

RESULTSThe total effective rate of 91.7% in the observation group was higher than that of 77.1% in the control group (P < 0.05). The dimensions of SF-36 of stress urinary incontinence patients were remarkably lower than those of normal group (all P < 0.05). The scores of ICI-Q-SF were decreased while the scores of SF-36 were increased obviously after treatment in both the observation group and the control group, there were pronounced improvements on physiological function, pain, physical activity, social function and affection function in the observation group (all P < 0.05).

CONCLUSIONThe quality of life for female stress urinary incontinence patients may be poor, however the dog-day acupuncture and tortoise-shell moxibustion combined with pelvic floor muscle exercises can improve the symptoms of urinary incontinence and increase the quality of life of patients.

Acupuncture Therapy ; Adult ; Aged ; Exercise Therapy ; Female ; Humans ; Middle Aged ; Moxibustion ; Muscle Contraction ; Muscles ; physiopathology ; Treatment Outcome ; Urinary Incontinence, Stress ; physiopathology ; therapy

OBJECTIVETo detect serve acute respiratory syndrome-associated coronavirus (SARS-CoV) and SARS-like-CoV in fruit bats captured in Guangzhou and its vicinity.

METHODSTotally 927 bats of 9 species (Cynopterus sphinx, Rousettus leschenaulti, Miniopterus schreibersi, Hipposideros pratti, Rhinolophusasinicus, Scotophilusakuhlii, Hipposideros Pomona, Rhinolophus affinis, and Rhinolophus pusillus) captured in Guangzhou and its vicinity from September 2004 to November 2005 were available for this investigation, from which 3,043 samples (813 throat swasb, 524 sera, 853 lung tissues and 853 colorectal tissue specimens) were obtained. SARS-Cov and SARS-like-CoV were detected in these specimens using diagnostic kit for novel coronavirus N protein (ELISA), SARS-CoV Virus RNA detection kit, fluorescence PCR, Genchip, RT-PCR and cell isolation culture methods.

RESULTS AND CONCLUSIONNo SARS-CoV and SARS-like-CoV were detected in the 3043 samples, indicating the current absence of SARS-CoV and SARS-like-CoV in the bats captured in Guangzhou and its vicinity.

Animals ; China ; epidemiology ; Chiroptera ; virology ; Disease Vectors ; Enzyme-Linked Immunosorbent Assay ; Humans ; Nucleocapsid Proteins ; metabolism ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; SARS Virus ; genetics ; isolation & purification ; metabolism ; Severe Acute Respiratory Syndrome ; epidemiology ; transmission ; virology

This study was aimed to investigate the occurrence and clinical significance of the SET-NUP214 fusion gene in patients with T-cell acute lymphoblastic leukemia (T-ALL), analyse clinical and biological characteristics in this disease. RT-PCR was used to detect the expression of SET-NUP214 fusion gene in 58 T-ALL cases. Interphase FISH and Array-CGH were used to detect the deletion of 9q34. Direct sequencing was applied to detect mutations of PHF6 and NOTCH1. The results showed that 6 out of 58 T-ALL cases (10.3%) were detected to have the SET-NUP214 fusion gene by RT-PCR. Besides T-lineage antigens, expression of CD13 and(or) CD33 were detected in all the 6 cases. Deletions of 9q34 were detected in 4 out of the 6 patients by FISH. Array-CGH results of 3 SET-NUP214 positive T-ALL patients confirmed that this fusion gene was resulted from a cryptic deletion of 9q34.11q34.13. PHF6 and NOTCH1 gene mutations were found in 4 and 5 out of 6 SET-NUP214 positive T-ALL patients, respectively. It is concluded that SET-NUP214 fusion gene is often resulted from del(9)(q34). PHF6 and NOTCH1 mutations may be potential leukemogenic event in SET-NUP214 fusion gene.
Carrier Proteins ; genetics ; Chromosome Deletion ; Chromosomes, Human, Pair 9 ; genetics ; Gene Expression ; Histone Chaperones ; genetics ; Humans ; Mutation ; Nuclear Pore Complex Proteins ; genetics ; Oncogene Proteins, Fusion ; genetics ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Receptor, Notch1 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transcription Factors ; genetics

The column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds from the EtOAc extract of medium and MeOH extract of cell cultures of Morus alba. Eight compounds were isolated. Based on physico-chemical properties and spectroscopic data, their structures were identified as isobavachalcone (1), genistein (2), norartocarpetin (3), albanin A (4), guangsangon E (5), mulberrofuran F (6), chalcomoracin (7), kuwanon J (8). Compounds 3-6 were isolated from the cell cultures of M. alba for the first time.
Benzofurans ; isolation & purification ; Cell Culture Techniques ; methods ; Chalcones ; isolation & purification ; Chromatography, Gel ; methods ; Chromatography, High Pressure Liquid ; Dextrans ; Genistein ; isolation & purification ; Morus ; chemistry ; cytology ; Plant Leaves ; chemistry ; cytology ; Plants, Medicinal ; chemistry ; cytology ; Silica Gel ; Terpenes ; isolation & purification