PTEN, a dual-specificity phosphatase, exerts its tumor-suppressive effects through the inhibition of cell cycle progression and cell immigration, therefore could be an important candidate for tumor-suppression. As study on prokaryotic expression of PTEN and its anti-tumor functions has not been reported, the present study aims at an efficient expression of functional PTEN in Escherichia coli and the investigation of its tumor-suppression activity. PTEN cDNA cloned in our lab previously was recombined into prokaryotic expression vector pET-44a(+) to construct pET-PTEN (pEP) and pET-Nus-PTEN (pENP). PTEN was fused with 6 x His tag in pEP, and with Nus in pENP, which could be useful for a stable and soluble expression. The recombinant vectors were transformed into both BL21 (DE3) (BL) and Rosetta-gami (DE3) pLysS (RG). The former is a normal expression host while the latter is optimized for expression of eukaryotic genes and folding of target proteins. On the induction of 0.5mmol/L IPTG, 55kD and 118kD specific protein bands were observed, corresponding to His-PTEN and Nus-PTEN fusion proteins, respectively. Western blot analysis showed the recombinant fusion proteins could react with PTEN polyclonal antibody. The recombinant HTEN was expressed both in soluble fraction and inclusion body. Higher expression levels of recombinant PTEN were obtained in BL (His-PTEN: 10.3%; NusA-PTEN: 18.7%), whereas the higher percentages of soluble recombinant proteins were observed in RG (His-PTEN: 4.7%; Nus-PTEN: 6.6%). The obtained recombinant fusion proteins were purified by affinity chromatography and were showed to be homogeneous in SDS-PAGE. In tumor-suppression experiments, His-PTEN was proved to have significant inhibition on growth of mice solid tumor with an inhibitory ratio of 58.76%, and on the proliferation of DU-145 tumor cells with an inhibitory ratio of 46.16%. The cell cycle progression of DU-145 tumor cells was also arrested from G0/G1 to S phase. His-PTEN from RG was proved to have significantly higher tumor-suppression activity than that from BL, indicating that there may be some advantages for eukaryotic genes to be expressed in the former host. This is the report of functional recombinant PTEN expressed in Escherichia coli. Purified His-PTEN was used for immunizing Kunming mice, and ascitic polyclonal antibodies raised against His-PTEN were generated using sarcoma 180 cells. At 1:2000 dilution, the antibodies could interact with PTEN by western blot. The present study has laid a foundation for application of PTEN in cancer therapy.
Animals ; Antibodies, Neoplasm ; biosynthesis ; immunology ; Antineoplastic Agents ; pharmacology ; Escherichia coli ; genetics ; metabolism ; Humans ; Liver Neoplasms, Experimental ; therapy ; Mice ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; immunology ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; pharmacology

Objective:To understand the current status of the level of chronic disease prevention and treatment literacy and health anxiety among chronic disease residents, as well as the urban-rural differences, in order to provide a basis for improving the level of chronic disease prevention and treatment literacy among chronic disease residents.Methods:This was a cross-sectional study. From July to August 2022, a multi-stage random sampling method was adopted to select 201 rural residents with chronic diseases in one rural health center and 242 urban residents with chronic diseases in two community health service centers. General demographic characteristics questionnaire, Chronic Disease Prevention and Control Literacy Questionnaire and Short version of Health Anxiety Scale were used for questionnaire survey.Results:There were 93 males and 108 females with chronic diseases in 201 rural chronic disease residents, and the age range was 19-69 years. There were 116 males and 126 females with chronic diseases in 242 urban chronic disease residents, and the age range was 18-69 years old. The score of chronic disease prevention and control literacy of rural chronic disease residents (7.86 ± 2.25) was lower than that of urban chronic disease residents (8.55 ± 2.03). The score of health anxiety of rural chronic disease residents (13.69 ± 5.26) was higher than that of urban chronic disease residents (11.67 ± 5.95). Both differences were statistically significant ( t=-3.43, 3.79, both P<0.05). After controlling the general demographic data, the layered linear regression analysis of rural chronic disease residents and urban chronic disease residents showed that health anxiety can negatively affect rural chronic disease residents and urban chronic disease residents of chronic disease prevention and treatment literacy level ( β=-0.185, -0.129, both P<0.05). Conclusions:There are urban-rural differences in chronic disease prevention and treatment literacy and health anxiety of chronic disease residents in community. It is necessary to strengthen psychological construction among rural chronic disease residents in order to improve their chronic disease prevention and treatment literacy.

OBJECTIVEWe propose an image-based key frames gating method for intravascular ultrasound (IVUS) sequence based on manifold learning to reduce motion artifacts in IVUS longitudinal cuts.

METHODSWe achieved the gating with Laplacian eigenmaps, a manifold learning technique, to determine the low-dimensional manifold embedded in the high-dimensional image space. A distance function was constructed by the low-dimensional feature vectors to reflect the heart movement. The IVUS images were classified as end-diastolic and non-end-diastolic based on the distance function, and the IVUS images collected in end-diastolic stage constitutes the key frames gating sequences.

RESULTSWe tested the algorithm on 13 in vivo clinical IVUS sequences (images 915±142 frames, coronary segments length 15.24±2.37 mm) to calculate the vessel volume, lumen volume, and the mean plaque burden of the original and gated sequences. Statistical results showed that both the vessel volume and lumen volume measured from the gated sequences were significantly smaller than the original ones, indicating that the gated sequences were more stable; the mean plaque burden was comparable between the original and gated sequences to meet the need in clinical diagnosis and treatment. In the longitudinal views, the gated sequences had less saw tooth shape than the original ones with a similar trend and a good continuity. We also compared our method with an existing gating method.

CONCLUSIONThe proposed algorithm is simple and robust, and the gating sequences can effectively reduce motion artifacts in IVUS longitudinal cuts.

Algorithms ; Angiography ; methods ; Artifacts ; Electrocardiography ; Humans ; Motion ; Reproducibility of Results ; Sensitivity and Specificity

OBJECTIVETo explore the morphometric abnormalities of brain gray matter (GM) in patients with chronic low back pain (CLBP).

METHODSThirty patients with CLBP and 30 healthy individuals were enrolled and examined with a 3.0 T magnetic resonance (MR) scanner. High-resolution T1 structural MR data were acquired and data analysis was performed using voxel-based morphometry (VBM) in FMRIB Software Library. The morphological differences were compared between the two groups.

RESULTSs Compared with the healthy control subjects, patients with CLBP showed decreased GM volumes in several brain cortical areas including the bilateral superior frontal gyrus, right frontal pole, left insular cortex, left middle and left inferior temporal gyrus (P<0.05, after TFCE correction). Increased GM volumes were found in the patients in the subcortical structures including the left thalamus, bilateral putamen, bilateral nucleus accumben and right caudate nucleus (P<0.05, after TFCE correction).

CONCLUSIONPatients with CLBP have different patterns of GM abnormalities in different brain regions, characterized by reduced GM volume in cerebral cortical regions and increased GM volume in the subcortical nuclei. Such changes might be associated with the maladaptation of the brain in chronic pain state.

Cerebral Cortex ; Frontal Lobe ; Gray Matter ; diagnostic imaging ; pathology ; Humans ; Low Back Pain ; physiopathology ; Magnetic Resonance Imaging ; Temporal Lobe ; Thalamus

BACKGROUNDEndogenous estrogen plays a very important role in the carcinogenesis and progression of breast cancer. The enzymes involved in the biosynthesis and metabolism of estrogen have been proposed to contribute to this effect. To examine this hypothesis, we conducted a case-control study to investigate the relationship between polymorphisms of genes responsible for estrogen biosynthesis (CYP17, cytochrome P450c17a and CYP19, aromatase cytochrome P450) and estrogen sulfation of inactivation (SULT1A1, sulfotransferase1A1) and the risk of breast cancer in Chinese women.

METHODSThis study involved 213 breast cancer patients and 430 matched controls. PCR-based restriction fragment length polymorphism (RFLP) and short tandem repeat polymorphism (STRP) assays were used to detect the mononucleotide transition of CYP17 and SULT1A1 and tandem repeat polymorphism of CYP19. Logistic regression analyses were used to determine OR and 95% CI of each and all three high-risk genotypes, of all three genotypes combined, and of estrogen exposure factors. The relationship between each high-risk genotype and clinicalpathological characteristics were also assessed.

RESULTSThe frequency of A2 allele of CYP17 was 49.8% in cases and 49.1% in controls (P = 0.82). The frequency of His allele of SULT1A1 was significantly higher in cases (13.6%) than in controls (9.5%) (P < 0.05). There was also significant difference of the (TTTA) 10 allele of CYP19 which was 12.4% in cases and 8.2% in controls (P < 0.05). When the CYP17 A2 allele, CYP19 (TTTA) 10 and SULT1A1 His allele were considered as the "putative high-risk" genotype, there was an increased risk of breast cancer with the number of high-risk genotypes in a dose-response effect (trend, P = 0.05). In multivariate analysis, the SULT1A1 genotype remained the most significant determinant for breast cancer, with OR = 2.37 (95% CI 1.23-4.74), followed by CYP19, with OR = 1.75 (95% CI 1.27-3.56). The (TTTA) 10 allele of CYP19 was associated with tumor size, and the His allele of SULT1A1 associated with status of lymph node metastasis.

CONCLUSIONSThis study supports the hypothesis that breast cancer can be initiated by estrogen exposure and that estrogen metabolizing genes are involved in this mechanism. This multigenic model is useful for identifying individuals who are at higher risks of breast cancer.

Adult ; Aged ; Aromatase ; genetics ; Arylsulfotransferase ; genetics ; Breast Neoplasms ; etiology ; genetics ; Case-Control Studies ; Estrogens ; metabolism ; Female ; Genetic Predisposition to Disease ; Humans ; Middle Aged ; Polymorphism, Genetic ; Risk Factors ; Steroid 17-alpha-Hydroxylase ; genetics

OBJECTIVETo investigate the differences in dielectric properties (relative permittivity and conductivity) between the mucosal surface and serosal surface of malignant colorectal tissues, adjacent tissues at 1 cm and 3 cm from the tumor focus and normal colorectal tissues.

METHODSThe dielectric properties of the mucosal and serosal surface of malignant colorectal tissues, adjacent tissues (1 cm and 3 cm) and normal colorectal tissues from 39 patients with colorectal cancer were measured with an open-ended coaxial probe within the frequency range of 50 MHz-3 GHz, and the corresponding dielectric properties were analyzed respectively; statistical tests of the data were used to analyze the dielectric properties at 6 specific frequency points.

RESULTSThe dielectric properties were significantly higher in the malignant mucosa surface than in the adjacent tissues and normal colorectal tissues at the 6 specific frequency points (P<0.01). The dielectric properties decreased progressively in adjacent tissues at 1 cm and 3 cm and normal mucosa surface. The mucosal and serosal surface of malignant tissues showed significant differences in dielectric properties at 64 MHz, 128 MHz, 298 MHz, 433 MHz, and 915 MHz (P<0.01) but not at 2450 MHz (P>0.01), but such differences were not observed in normal tissues (P>0.01).

CONCLUSIONThe dielectric properties of the mucosal surface of the mucosal decrease in the order of malignant colorectal tissue, adjacent tissues at 1 cm and 3 cm from the tumor foci and normal colorectal tissues. The dielectric properties are higher in the mucosal surface than in the serosal surface in the malignant tissue, but comparable in normal colorectal tissues.

OBJECTIVETo develop a near-infrared fluorescence imaging system based on the fluorescence properties of methylene blue.

METHODSAccording to the optical properties of methylene blue, we used a custom-made specific LED light source and an interference filter, a CCD camera and other relevant components to construct the near-infrared fluorescence imaging system. We tested the signal-to-background ratio (SBR) of this imaging system for detecting methylene blue under different experimental conditions and analyzed the SBR in urine samples collected from 15 Wistar rats with intravenous injection of methylene blue at the doses of 0, 1.4, 1.6, 1.8, or 2.0 0 mg/kg methylene blue.

RESULTSThe SBR of this imaging system for detecting methylene blue was affected by the concentration of methylene blue and the distance from the sample (P<0.05). In the urine samples from Wistar rats, the SBR varied with the the injection dose, and the rats injected with 1.6 mg/kg methylene blue showed the highest SBR (8.71∓0.20) in the urine (P<0.05).

CONCLUSIONThis near-infrared fluorescence imaging system is useful for fluorescence detection of methylene blue and can be used for real-time recognition of ureters during abdominal surgery.

OBJECTIVETo investigate the effects of Vaspin on lipopolysaccharide (LPS)-induced acute respiratory distress syndrome (ARDS) in mice and explore the possible mechanism.

METHODSForty male C57B/L6 mice were randomized equally into control group, LPS group, Vaspin group and wortmannin group with corresponding treatments. The pathological changes of the lung tissues were evaluated by HE staining, and the severity of pulmonary edema was measured according to the wet/dry ratio (W/D) of the lung tissue. The lung permeability was evaluated by detecting total protein concentrations in the bronchoalveolar lavage fluid (BALF) using bicinchoninic acid (BCA) assay. Myeloperoxidase (MPO) activity in the lung tissue was detected using a MPO assay kit, and the levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) in the lungs were measured using ELISA. Immunohistochemical staining was performed to detect the expression of vascular cell adhesion molecule-1 (VCAM-1) and Western blotting was used to detect the protein expressions of cleaved caspase-3 and p-Akt in the lung tissues.

RESULTSCompared with the control group, the mice in LPS group displayed typical ARDS pathological changes in the lungs with significantly increased W/D, total protein concentrations in BALF, lung MPO activity, levels of IL-1β and TNF-α, and pulmonary expressions of VCAM-1 and cleaved caspase-3 (P<0.05) but decreased expression of p-Akt (P<0.05). These changes induced by LPS were significantly alleviated by the administration of Vaspin (P<0.05). The protective effects of Vaspin against ARDS were obviously attenuated by the PI3K inhibitor wortmannin (P<0.05).

CONCLUSIONVaspin protects against LPS-induced ARDS in mice possibly by inhibiting inflammation and protecting vascular endothelium through upregulation of the PI3K/Akt signal pathway.

OBJECTIVE: To investigate the prevalence and gene distribution of thalassemia among people at reproductive age in yuzhong district, Chongqing. METHODS: 1000 pre -pregnancy examination couples in yuzhong district were investigated. Peripheral venous blood was extracted and next-generation sequencing was used to screen the thalassemia genes. RESULTS: Among the 1000 pregnant couples, the thalassemia gene carrying rate was 7.45%, the carrying rate of α and β thalassemia genes were 4.60% and 2.10%, respectively. The most common α thalassemia genotypes in αα/-α3.7 (53.26%), αα/--SEA (23.91%), αα/-α4.2 (11.96%); and the most common genotypes in β thalassemia genotypes were mainly Codons17 (A＞T) (26.19%)、Condon41/42 (-TTCT) (26.19%)、IVS-II-654 (C＞T) (14.29%) At the same time, 3 cases of α and β complex thalassemia and 3 pairs of homotypic thalassemia genes were detected, more over, 12 cases of 5 new genes were found. CONCLUSION Yuzhong district of Chongqing is a high incidence area of thalassemia, and the diversity of gene mutation types is relatively rich. Screening for thalassemia before pregnancy is of great significance to improve the quality of population.
China ; Female ; Genotype ; High-Throughput Nucleotide Sequencing ; Humans ; Mutation ; Pregnancy ; Prevalence ; alpha-Thalassemia ; beta-Thalassemia

OBJECTIVE: To investigate the clinical characteristics of myelodysplastic syndrome (MDS) patients with chromosome 21 karyotype abnormality. METHODS: The clinical data of 155 patients with MDS were retrospectively analyzed, the clinical characteristics, survival and factors affecting prognosis of chromosome 21 karyotype abnormality patients were analyzed. RESULTS: Among 155 MDS patients, 4 were 5q- syndrome, 41 were MDS-EB-I, 35 were MDS-EB-II, 27 were MDS-SLD, 46 were MDS-MLD, 1 was MDS-RS-SLD, and 1 was MDS-U. The median follow-up time was 11.0(0.1-120.9) months. Among 155 MDS patients, 13 (9.0%) showed chromosome 21 abnormalities. Among the 13 patients with chromosome 21 karyotype abnormalities, there were 5 cases with simple +21 karyotype, 1 case with del (21q12), 1 case with +8, +21, 1 case with i(21q), 1 case with 20q-, +21, and 4 cases with complex karyotype involving chromosome 21; including 2 cases of MDS-SLD, 4 cases of MDS-MLD, 5 cases of MDS-EB-I and 2 cases of MDS-EB-II. The median survival time of the patients was 3.1 (0.1-6.7) months. CONCLUSION Chromosome 21 karyotype abnormality is rare in MDS, and the prognosis is worse than the patients without chromosome 21 abnormalities.
Chromosomes, Human, Pair 21 ; Humans ; Karyotype ; Karyotyping ; Myelodysplastic Syndromes/genetics* ; Retrospective Studies