Gene therapy has emerged as an efficient modality to treat human diseases.This method is based on the transfer of genetic material to tissues to induce a curative effect.Gene therapy vectors are molecular constructs used to facilitate the penetration of genomic sequences inside the cells.Viral vectots have however several limitations when administered directly to the patient.They may cause significant toxicity by activating innate immunity or by eliciting an adaptive immune response against viral proteins.In addition,targeting the vector to the desired site is an issue when given systemically.The use of cells as vehicles for gene therapy vectors has many advantages.The combination of cell-viro-gene therapy has been thought as a new and promising strategy for therapy of cancer.The targeting vector to cancer stem cells will become a new direction in the field of gene therapy.In this article,we will introduce progressions,limitations and future directions of gene therapy of liver cancer.

Objective To analyze CMV DNA stability of 30 EDTA plasma samples in the order of magnitude between 300 and 100 000 copies/mL over a 21 day period. Methods Thirty plasma samples were grouped into three categories according to the CMV DNA loads , including low CMV DNA contents , intermediate CMV DNA loads and high CMV DNA loads. Ten milliliters of whole blood was freshly collected from each patient. Plasma samples without hemolysis were divided into 1-ml aliquots. One aliquot was processed immediately (Day 0) for baseline PCR assays. The remaining aliquots were then processed after one , two, three, seven, 14 or 21 day of storage at 4℃. Results There was no significant difference between the mean of the difference time point in viral loads following storage at 4 ℃ by paired-samples t test, including Day 1 compared to Day 0 (t = 1.654, P =0.109), Day 2 compared to Day 0 (t = 1.487, P = 0.148), Day 3 compared to Day 0 (t = 1.609, P = 0.118), Day 7 compared to Day 0 (t=0.831, P=0.413), Day 14 compared to Day 0 (t=1.721, P=0.096), and Day 21 compared to Day 0 (t=0.244, P=0.810). Conclusion The concentration of CMV DNA in all samples stored at 4 ℃ for 21 days did not differ significantly from the baseline viral load ,and it was not observed the trend in continued degradation in different time point (Day 1, 2, 3, 7 and 14).

OBJECTIVE: To determine the regulation effect of bone morphogenetic protein-4 (BMP-4) on the proliferation and differentiation of rat hepatic precursor cells. METHODS: We used Noggin (200 ng/mL) as the function blocking control of BMP-4, and the hepatic precursor cells of WB-F344 were treated with recombinant BMP-4 at 50 ng/mL at different time points. The proliferation of WB-F344 cells were tested by methyl thiazolyl tetrazolium (MTT) colorimetric assay. The ultrastructural characters of differentiated WB-F344 cells regulated by BMP-4 were observed under a transmission electron microscope. RT-PCR was used to examine mRNA expression of specific molecular markers for different cellular phenotypes potentially differentiated from the WB-F344 cells. RESULTS: At different time points, the absorbance values in the BMP-4 treatment groups were higher than those in the control groups of Noggin and blank treatment (P<0.01). The WB-F344 cells treated with BMP-4 exhibited typical ultrastructural characters of well-differentiated epithelial cells. The hepatocyte mRNA markers were more significantly promoted in the differentiated WB-F344 cells in the BMP-4 treatment group than those in the other 2 control groups. CONCLUSION BMP-4 can promote the proliferation and directional differentiation towards hepatocytes of rat hepatic precursor cells of WB-F344.
Animals ; Bone Morphogenetic Protein 4 ; antagonists & inhibitors ; genetics ; physiology ; Carrier Proteins ; pharmacology ; Cell Differentiation ; Cell Line ; Cell Proliferation ; Hepatocytes ; cytology ; Rats ; Recombinant Proteins ; Stem Cells ; cytology

OBJECTIVE: To determine the treatment effects of transplanted hepatic progenitor cells (WB-F344 cells) combined with heparin on the acute liver injury in SD rats. METHODS: A total of 2*10(7) hepatic stem cells (WB-F344) infected with GFP lentivirus and 8 μL heparin were transplanted through the spleen in SD rats with acute liver injury, which was induced by an intraperitoneal injection of CCl4. The liver and spleen tissues underwent fluorescence examination 1 day after the transplantation. The liver functions were tested, and the liver tissues were histopathologically examined on the 3rd, 7th, 14th, and 28th day of the cell transplantation. RESULTS: The transfected WB-F344 cells expressed GFP 3 days after the lentivirus infection and were found in the rat liver 1 day after the WB-F344 transplantation. The liver function and histopathological recovery of the liver tissues in the group of WB-F344 transplantation were better than those of the control group (P<0.05). CONCLUSION Transplantation of hepatic stem cells combined with heparin can promote the liver recovery in rats with acute liver injury induced by CCl4.
Animals ; Carbon Tetrachloride ; Carbon Tetrachloride Poisoning ; Heparin ; therapeutic use ; Hepatocytes ; transplantation ; Liver Failure, Acute ; chemically induced ; therapy ; Male ; Rats ; Rats, Sprague-Dawley ; Stem Cell Transplantation ; methods

Objective:To analyze the consistency and correlation between the Chinese Version of Swanson Nolan and Pelham, Version Ⅳ Scale (SNAP-Ⅳ) and the Integrated Visual and Auditory Continuous Performance Test (IVA-CPT) in the assessment of attention deficit hyperactivity disorder (ADHD), thus providing a reliable basis for the diagnosis of ADHD, and reducing the misdiagnosis rate and missed diagnosis rate.Methods:Clinical data of children to be diagnosed as ADHD in the Department of Children′s Rehabilitation, Yuying Children′s Hospital of the Second Affiliated Hospital of Wenzhou Medical University from October 2019 to July 2020 were collected.A total of 282 SNAP-Ⅳ and IVA-CPT profiles were collected, and the Kappa test and Pearson test were used to retrospectively analyzed for their consistency and correlation in the diagnosis of ADHD. Results:SNAP-Ⅳ and IVA-CPT were consistent in the diagnosis of ADHD (Kappa value=0.514, total coincidence rate=65.6%, P<0.000 1). Inattention subset scores of SNAP-Ⅳ were consistent with the assessment of ADHD by IVA-CPT (Kappa value=0.485, total coincidence rate=75.5%, P<0.000 1). Inattention subset scores of SNAP-Ⅳ were negatively correlated with the Full Scale Attention Quotient (FAQ) in IVA-CPT ( r=-0.71, P<0.000 1). Hyperactivity-impulsive subscale in the SNAP-Ⅳ and IVA-CPT were consistent in the assessment of hyperactivity-impulsive behavior (Kappa value=0.585, total coincidence rate=81.6%, P<0.000 1). Hyperactivity-impulse subset scores were negatively correlated with the Full Scale Response Control Quotient (FRCQ) in IVA-CPT ( r=-0.74, P<0.000 1). Conclusions:Both SNAP-Ⅳ and IVA-CPT have certain diagnostic potential of ADHD, showing good consistency and correlation.They can be both used to provide a more comprehensive diagnosis basis, thereby reducing the misdiagnosis rate and missed diagnosis rate of ADHD.

OBJECTIVETo understand the prevalence of thyroid diseases and its influencing factors of iodine on thyroid gland function and autoimmune among fertile women in different iodine intake areas.

METHODSCross-sectional method was used for descriptive epidemiology. 236 women aged 19 to 45 years were sampled in 2011, in Shanxi province. Questionnaire was used to include general data on place, name, age etc. Sample of water from home, one time random urine sample and venous blood were collected to test the iodine contents using arsenic and cerium catalysis spectrophotometric methods. Finally, in blood, free triiodothyronine (FT3), free thyroxine (FT4), thyrotrophin (TSH) in blood were tested under auto-CLIA and anti-thyroid peroxidase (anti-TPO), anti-thyroglobulin (anti-TG) through radio-immunological methods.

RESULTS1)The urine iodine's medians were 486.9 µg/L for fertile women in high iodine areas, and 192.6 µg/L in low iodine areas, with difference on urine iodine level statistically significant (Z = -10.676, P = 0.000). 2) Levels of blood FT3 and FT4 in women from high iodine areas were obviously lower than those from proper iodine areas(t = -2.884, P = 0.004; t = -2.862, P = 0.005), but the level of TSH in high iodine areas was higher than that of proper iodine areas(t = 2.332, P = 0.021). 3) In both areas, the rate of the thyroid dysfunction with positive antibodies was obviously higher than those with negative antibodies (χ² = 20.941, P = 0.000;χ² = 5.596, P = 0.018), while the rate of the thyroid dysfunction with positive antibodies and the level of TSH in the blood for high iodine women higher than those in women with proper iodine level(χ² = 5.708, P = 0.37;t = -2.177, P = 0.031). 4)The morbidity rate of inferior clinical hyperthyroidism for women in high iodine areas was obviously higher than those in proper iodine areas(χ² = 9.542, P = 0.003), while the morbidity rate of inferior clinical hypothyroidism for women with positive antibodies in two areas obviously higher than those with negative antibodies (χ² = 17.264, P = 0.000; χ² = 6.002, P = 0.044).

CONCLUSIONMorbidity rate of inferior clinical hypothyroidism for women in high iodine areas was obviously higher than those in proper iodine areas, suggesting that there were potential risks of hypothyroidism for overdose iodine intake which causing the existence of positive thyroid antibodies. Monitoring programs on iodine nutrition and thyroid function among women living in high iodine areas should be strengthened.

Adult ; China ; epidemiology ; Female ; Humans ; Iodides ; administration & dosage ; Iodine ; urine ; Middle Aged ; Nutritional Status ; Prevalence ; Thyroid Diseases ; epidemiology

Corynebacterium glutamicum, an important microorganism to produce amino acids and organic acids, has been widely applied in food and medicine fields. Therefore, using editing tools to study the function of unknown genes in C. glutamicum has great significance for systematic development of industrial strain with efficient and novel production capability. Recently, gene editing has been greatly developed. Traditional gene editing based on homologous recombination and gene editing mediated by nuclease are successfully applied in C. glutamicum. Among these, the CRISPR system has been developed to be a main tool used for gene knockout of C. glutamicum due to its advantages of efficiency, simplicity and good target specificity. However, more efficient and reliable knockout system is still urgently demanded, to help develop high-performing strains in industrial application.
CRISPR-Cas Systems ; Clustered Regularly Interspaced Short Palindromic Repeats ; Corynebacterium glutamicum ; genetics ; Gene Editing ; trends ; Glutamic Acid ; Industrial Microbiology ; trends

Steroid saponins are secondary metabolites with multiple medicinal values that are found in large quantities in natural medicines, especially Vernonia amygdalina, a famous nature medicine for the treatment of tonsillitis, diabetes, pneumonia. The current study was designed to combine molecular networking (MN) with diagnostic ions for rapid identification of Δ^7,9(11) stigmastane-type saponins which were the α-glucosidase inhibitory active substances in V. amygdalina. First, the α-glucosidase inhibitory activities of five Δ^7,9(11) stigmastane-type steroid saponins that were previously isolated were screened, which indicated that the Δ^7,9(11) stigmastane-type steroid saponin was one of the active constituents responsible for ameliorating diabetes. Furthermore, a strategy was proposed to identify stigmastane-type steroid saponins and verify the plausibility of derived fragmentation pathways by applying MN, MolNetEnhancer and unsupervised substructure annotation (MS2LDA). Based on this strategy, other seven Δ^7,9(11) stigmastane-type steroid saponins were identified from this plant. Our research provide scientific evidence for the antidiabetic potential of the steroid saponin-rich extract of V. amygdalina leaf.
alpha-Glucosidases/metabolism* ; Vernonia/chemistry* ; Plant Extracts/chemistry* ; Plant Leaves/chemistry* ; Saponins/chemistry* ; Steroids/chemistry* ; Diabetes Mellitus