AIM:To construct eukaryotic expression vector expressing double shRNA sections targeting Survivin gene.METHODS: Eukaryotic expression vector expressing double shRNA sections targeting Survivin gene were designed and chemically synthesized.They were directionally inserted into plasmid pGenesil-1 with respectively U6 promoter and termination code,the common green fluorescence protein(EGFP) gene and Neo gene. In this way,the vector of pGenesil-1 shRNA containing 2 sections of Survivin shRNA were constructed and they were transfected into the pancreatic cancer cell Bx-PC3.Transfection was detected by fluorescence microscope.The inhibition expression of Survivin mRNA was measured by RT-PCR.RESULTS: HE1 and HE2 plasmids were identified by the biocatalyst cut which confirmed the exactitude and were analyzed by the sequence analysis which verified the perfect clone plasmid inserted by them.CONCLUSION: A eukaryotic expression vector of double short hairpin RNA for Survivin gene is successfully constructed.The pancreatic cancer cells Bx-PC3 succeed to be transfected and expression of Survivin mRNA is inhibited obviously.

Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoembryonic Antigen ; blood ; Catheter Ablation ; methods ; Colorectal Neoplasms ; pathology ; Combined Modality Therapy ; Fluorouracil ; therapeutic use ; Humans ; Leucovorin ; therapeutic use ; Liver Neoplasms ; blood ; drug therapy ; secondary ; surgery ; Neoplasm Recurrence, Local ; Organoplatinum Compounds ; therapeutic use

Humans ; Neoplasm Metastasis ; pathology ; Neoplasm Staging ; methods ; Neoplasms ; classification ; pathology ; therapy

A new ionizing-radiation resistant strain WGR702 was isolated from arid soils which had been ra-diated. The strain WGR702 was Gram-positive and coccus, the diameter of the cell was 1.5 ?m～2.5?m.The strain WGR702 was pink-pigmented, motile, facultative anaerobe and non-spore forming. The range tem-perature and pH for strain WGR702 growth were 10℃～35℃ and pH 5.0～10.0 respectively. The strain WGR702 had a G+C content of 60.5 mol%. UV and gamma radiation survival curves showed the strain WGR702 had highly ionizing-radiation resistant. Phylogenetic analysis of the 16S rDNA gene sequences (EU315117) showed 94.79%～98.53% similarities with other recognized Serratia species. Primary charac-teristics that distinguish isolate WGR702 from the species of genus Serratia include the cells are spherical and Gram-positive. Based on the phenotypic, biochemical and physiological characteristics differences it is proposed that the new isolated strain WGR702 might be classified as a novel species of Serratia.

Objective: To examine the effect of early comprehensive interventions on the physical growth and nerve development among premature infants, so as to provide insights into the follow-up management of premature infants after discharge from hospital. Methods: A total of 130 premature infants delivered in Shaoxing Municipal Maternal and Child Health Care Hospital from 2019 to 2021 were selected and divided into high- and low-risk groups according to gestational age and birth weight, while 306 full-term normal infants in the same hospital during the study period served as controls. All premature infants were given early comprehensive interventions until age of 12 months, including nutritional support and parental guidance of children's feeding and development, and all normal infants received periodical health checkup according to the basic public health service program. All infants received periodical measurements of height, weight and head circumference, and the 12-month intellectual and motor development ability was measured using the Bayley Scales of Infant Development revised in Chinese cities was used to assess. Infants' physical growth, mental development index (MDI) and motor development index (PDI) were compared among groups. Results: There were 130 premature infants with gestational ages of 28 to 36 weeks and birth weight of 1 200 to 3 440 g, including 79 male infants, and there were 80 infants in the low-risk group and 50 infants in the high-risk group. The full-term infants had a gestational age of 37 to 42 weeks, and birth weights of 2 500 to 4 000 g, including 162 male infants. There were significant differences in height (Wald χ2=28.664, P<0.001) and head circumference growth (Wald χ2=19.312, P=0.013) among the three groups as revealed by the generalized estimating equation; however, no significant differences were seen in the 12-month weight (F=0.639, P=0.528), height (F=1.051, P=0.350) or head circumference (F=0.318, P=0.728) among the three groups. The percentages of abnormal MDI were 2.00%, 0 and 1.31% among the high-risk premature infants, low-risk premature infants and full-term infants at ages of 12 months (χ2=1.319, P=0.517), while the percentages of abnormal PDI were 20.00%, 7.50% and 5.56% among the three groups at ages of 12 months (χ2=12.818, P=0.002). Conclusions Following implementation of early comprehensive interventions, the premature infants have favorable physical growth and comparable MDI with full-term infants; however, a high percentage of abnormal PDI is seen in high-risk premature infants. An improvement in the motor development among high-risk premature infants is recommended to be emphasized during the management of premature infants.

This study started from the effect of baicalin (BC), the main active component of the labiaceae plant Scutellaria baicalensis, on collagen-induced arthritis (CIA) in rats, to explore the mechanism of glucose metabolism reprogramming in fibroblast like synoviocytes (FLSs), a key effector cell of synovial inflammation in rheumatoid arthritis (RA). First of all, CIA rats and tumor necrosis factor-α (TNF-α)-induced RASFs in vitro and in vivo models were established, the arthritis index (AI) score and histopathological changes of CIA rats after BC administration were observed, and the levels of inflammatory factors in serum and cell supernatant were quantified by ELISA, immunocytochemistry and Western blot were used to detect the expression of G-protein-coupled receptor 81 (GPR81) and pyruvate dehydrogenase kinase 1 (PDK1) proteins. In addition, the kit was used to measure the levels of key products and enzyme activities in glucose metabolism reprogramming. The results showed that BC (50, 100 and 200 mg·kg^-1) could alleviate the symptoms of arthritis in CIA rats in a dose-dependent manner, inhibit synovial hyperplasia, alleviate the infiltration of inflammatory cells, down-regulate the levels of pro-inflammatory factors TNF-α and interleukin (IL)-1β, and up-regulate the levels of anti-inflammatory factor IL-10 in CIA rats. At the same time, the secretion levels of lactate, pyruvate, acetyl-CoA, citrate and the activity of lactate dehydrogenase B (LDH-B) were decreased, and the expressions of GRP81 and PDK1 were down-regulated, suggesting that BC mediated the reprogramming process of glucose metabolism. However, when GPR81 inhibitor 3-OBA inhibited lactate uptake, the activity of LDH-B was significantly increased, suggesting that BC inhibited the expression of PDK1, a key enzyme in the reprogramming metabolism from glycolysis to oxidative phosphorylation. All animal experiments in this study were conducted in accordance with the ethical standards of the Laboratory Animal Care Center of Anhui University of Chinese Medicine (approval number: AHUCM-rats-2021049). These studies revealed that baicalin mediated metabolic reprogramming of RASFs from glycolysis to oxidative phosphorylation by inhibiting PDK1 protein expression, and alleviated joint inflammation in CIA rats.

Objective:To summarize the characteristics of genetic variation and prenatal diagnosis in pedigrees with X-linked adrenoleukodystrophy (X-ALD) and elucidate the value of prenatal diagnosis in preventing the birth of children with X-ALD.Methods:Twenty pedigrees, clinically diagnosed with X-ALD in Peking University First Hospital from November 2012 and March 2019, were included in this retrospective study. Genomic DNA was extracted from peripheral blood and amniotic fluid or chorionic villi samples of probands and their families for detecting variants in ATP-binding cassette subfamily D member 1 ( ABCD1) gene using polymerase chain reaction (PCR)-Sanger sequencing. Linkage analysis was also performed on five microsatellite markers near ABCD1 gene to exclude maternal contamination. Characteristics of ABCD1 gene variants and prenatal diagnosis of X-ALD pedigrees were summarized by descriptive statistics. Results:Twenty ABCD1 gene variants were identified in the 20 pedigrees. The variants in three probands that were not detected by next-generation sequencing were identified by PCR-Sanger sequencing. Among the mothers of the 20 probands, 17 carried ABCD1 variants and three did not. We performed 24 prenatal diagnoses on 20 pregnancies (24 fetuses) and identified eight fetuses with variants who were finally terminated. The 16 cases without variants were born alive. The validation results obtained after termination or delivery were consistent with those performed prenatally. Conclusions:No hotspot variants in ABCD1 gene are detected in these X-ALD patients and most variants are maternally inherited. PCR-Sanger sequencing is an effective method for detecting ABCD1 variants. Prenatal diagnosis for mothers who had a body with X-ALD could prevent another one from birth.

Objective:To investigate the molecular interaction between non-structural protein 3 serine protease of hepatitis C virus(HCV)and wild type P53,and to lay the basis for elucidating the mechanism of oncogenesis of hepatocellular carcinoma(HCC)after infection of HCV.Methods:The recombinant plasmids,pGAD424-NS3,pGAD424-NS315aa- and pGAD424-NS330aa-,were constructed and the interaction between NS3 serine protease and its cofactor NS4A,the interaction between wild type P53 and NS3 serine protease and its N-truncated mutants were dectected qualitatively and quantitatively in yeast two-hybrid system.Results:The results indicated that interaction existed not only between full-length NS3 serine protease and P53,but also between N-truncated mutants of NS3 serine protease and P53.Furthermore,the difference between enzyme activity unit(IU)of β-gal induced by these interactions was not significant(P>0.05).Conclusions:NS3 serine protease of hepatitis C virus and its N-truncated mutants can interact with wild type P53,and the region of NS3 serine protease involved in the interaction may be located in its C-terminal,but not in its N-terminal.

Objective　To observe the effects of residual perfluorocarbon liquid(PFCL) droplets in the anterior chamber in patients after vitreoretinal(VR) surgery for complicated retinal detachments.Methods　The study group consisted of nine patients(nine eyes)aged 16～64 in whom droplets of perfluorodeclin liquid(PFDL) appeared in the anterior chamber subsequent to VR surgery for complicated retinal detachments involving scleral bucking, vitrectomy and/or lensectomy, the peeling of pre-retinal membrane, perfluorodecalin injection, and perfluorodeclin liquid-silicone oil or fluid-air exchange. Mean follow-up was 10.3 months.Results　Three to 30 droplets of perfluorodeclin liquid were found in the anterior chambers in 9 eyes. There was no corneal complication or inflammatory reactions or blood vessel invasion into the corneal stroma or increased intraocular pressure.Conclusion　Residual perfluorodeclin liquid droplets in the anterior chamber are well tolerated and do not induce corneal damage or ocular inflammation.

Objective:To investigate the effects of sensitized sera on bone marrow transplantation and clarify the role of antibody in the process of rejection.Methods:Two hundred microlitres sensitized sera or non-sensitized sera were injected into normal BALB/c one day before transplantation.Ten millions (1?107) bone marrow cells from C57BL/6 were transfused to the recipients after lethal irradiation.The donor-reactive antibodies in recipients before transplantation were measured by complement-dependent cytotoxicity (CDC) method.Moreover,the survival analysis and engraftment evaluation of the recipients were carried post transplantation.Results:The CDC results showed that donor-reactive antibodies existed in the recipients which had received sensitized sera transfusion.Eighty percent (80%) of the recipients received sensitized sera transfusion died of bone marrow failure about 10 days post transplantation,while the recipients received non-sensitized sera transfusion were long-term alive.Furthermore,the hematopoietic recovery and percentage of donor chimera analysis declined along with time in the sensitized sera transfusion recipients,and there were significant differences compared with those in the non-sensitized sera transfusion recipients (P