Objective: To explore the effect of artesunate (Art) on Akt/GSK-3β/β-catenin signal pathway.Methods: Art at different concentrations (0, 12.5, 25, 50 μg·ml-1) was used to treat human hepatic stellate cells (LX-2), and CCK-8 assay was used to detect the cell proliferation to determine the optimal concentration.Art inhibitor (MK-2206) at different concentrations (0~8 μmol·L-1) was given to LX-2 cells, and a Western Blot method was applied to determine the optimal inhibition concentration.Art, MK-2206 and MK-2206+Art were respectively given to LX-2 cells, and a Western Blot method was used to detect the levels of Akt, p-Akt, GSK-3β, p-GSK-3β and β-catenin proteins.Results: CCK-8 assay was used to detect the cell survival rate, and the survival rate was 80% after the 24-hour treatment with 25 μg·ml-1 Art.The results of Western Blot showed that MK-2206 at 6 μmol·L-1 could effectively inhibit the expression of p-Akt.Compared with those of the control group, the levels of Akt, p-Akt, p-GSK-3β and β-catenin protein were significantly different (P＜0.05) in Art (25 μg·ml-1) group, MK-2206 (6 μmol·L-1) group and MK-2206 (6 μmol·L-1) + Art (25 μg·ml-1) group.The expression of GSK-3β and Akt in MK-2206+Art group had no significant difference when compared with that in Art group and MK-2206 group (P＞0.05), while the levels of p-Akt, p-GSK-3β and β-catenin were significantly reduced (P＜0.01).Conclusion: Art exhibits the influence on the relative factors in Wnt/β-catenin signal pathway by Akt/β-catenin, subsequently inhibits the cell proliferation and alleviates the liver fibrosis process.

Objective:To explore the effect of P-glycoprotein ( P-gp) activity on its mediated imatinib mesylate accumulation and intracellular drug membrane permeability. Methods: 1199G/wt ABCB1 and 1199A/mut recombinant plasmids were transferred into HEK293 cells, respectively, and the expression levels of mRNA in different cell models were investigated by RT-PCR. Cell Counting Kit-8(CCK-8) assay was used to detect the drug toxicity in different cell models, HPLC was applied to determine the drug concentra-tion in different cell models and evaluate the intracellular accumulation, and transmembrane resistance experiment was employed to de-tect the transmembrane permeability and evaluate the effect of P-gp activity on drug transportation. Results: Cytotoxicity test showed that the drug concentration in the transferred cells was lower than that in the control group, which proved that P-gp had the function of mediating drug out of cells. HPLC and transepithelial electrical resistance experiment showed that compared with the wild type of AB-CB1 (1199G) cells, mutation of ABCB1 1199A cells had stronger effect on P-gp mediated mesylate imatinib accumulation and drug membrane permeability. Conclusion:The experiment manifested that ABCB1 (1199G/A) site mutation can change the coding protein P-gp activity and the polymorphisms will lead to the increase of mesylate imatinib clearance rate and the decrease of effective drug con-centration in target cells. Meanwhile, the clarification of ABCB1 genetic types in clinics can guide the individualized medication of imatinib mesylate.

Objective To observe the clinical effect of gonadotropin releasing hormone analogues in treatment of the girls with idiopathic central precocious puberty(ICPP). Methods Twelve girls with ICPP received therapy with GnRH-A.Before and after the end of treatment for 6 months,secondary sexual characteristics and growth velocity were observed,pelvic ultrasonic examination,X-ray bone age(BA),serum level of E_2 and LHRH stimulating test were detected,and the predicted adult height(PAH) was analysed. Results After treatment,the size of breasts,uterus and ovaries were significantly reduced.The basic and peak levels of serum LH and FSH by LHRH stimulation were also significantly reduced.The ratio of BA/CA(chronology age)was decreased.The PAH was increased. Conclusions GnRH-A can effectively depress the sexual characteristics,reduce the maturation of BA,and also improve the PAH in girls with ICPP.

Objective To prepare loratadine nanoparticle suspension and investigate the stability of it. Methods Loratadine nanoparticle suspension was prepared by anti-tumor agent precipitation method,the nanosuspension was characterized by particle size analyzer and transmission electron microscopy, the optimal prescription was screened, and the stability of nanosuspension was investigated by HPLC. Results The optimal prescription stablizer was SDS and organic phase was ethanol. The drug loading radio was 1:2 and the proportion of organic phase to water was 5:10 and the time of high shear was 5 min. Loratadine suspension was pale blue with a uniform emulsion.The nanoparticles were spherical,with an average particle size of 112.8 nm,the PDI of 0.095 and the Zeta potential of -38.6 mV.The suspension had the best physical and chemical stability at room temperature. Conclusion The preparation method of loratadine suspension with good stability is simple, and it’s expected to become the new nano-drug delivery system of loratadine.

OBJECTIVE: To summarize the clinical repairment experience of postoperative skin defects in head and neck cutaneous tumor. METHOD: From July 2002 to September 2006, 24 patients with head and neck cutaneous tumor were treated in our department. Every specimen and resection margin of all cases were monitored by intraoperative frozen section. Extents of operative skin defects were from 1.0 cm x 1.5 cm to 3.0 cm x 3.5 cm, all of which were repaired by rhomboid flap. RESULT: All the cases were primarily repaired, and followed up 6 month to 3 years. The repaired skins had the normal colour, without obviously scars or secondary deformations. CONCLUSION The rhomboid flap is reasonably designed, conveniently procured and manipulated. It is a better method to immediately repair the head and neck skin defects after radical excision of tumor.
Adult ; Aged ; Female ; Head and Neck Neoplasms ; surgery ; Humans ; Male ; Middle Aged ; Postoperative Period ; Reconstructive Surgical Procedures ; methods ; Skin Neoplasms ; surgery ; Skin Transplantation ; methods ; Surgical Flaps

To establish the amplification-refractory mutation system quantitative real-time PCR (ARMS-qPCR) method based on qPCR technique for detecting the c.2T>C mutation of SLC25A13 gene and validate its diagnostic performance. According to the principle of ARMS-qPCR primer design, the specific primers were designed for the conserved sequence of SLC25A13. The c.2T>C mutation ARMS-qPCR detection assay of SLC25A13 gene and the corresponding Sanger sequencing system were established through the use of the synthetic plasmids of homozygous mutation and 200 human peripheral blood specimens which were verified by Sanger sequencing as templates, and the diagnostic efficacy of the qPCR assay was validated by using nucleic acid extracted from another 200 human peripheral blood specimens and the results obtained were compared with the Sanger sequencing results as the gold standard, and the consistency of the two detection methods was analyzed. The results showed that the qPCR assay could accurately identify artificial plasmids carrying different mutations of SLC25A13 gene, and distinguish between wild type SLC25A13 gene and the c.2T>C mutation. This method was used to detect the mutation status of SLC25A13 c.2T>C in human peripheral blood, and the detection results were 100% consistent with the Sanger sequencing results. Among the 200 blood samples, 8 samples (4%) carried the c.2T>C mutation of SLC25A13 gene and 192 samples (96%) did not carry it. In conclusion, the ARMS-qPCR test established in this study can quickly, simply and accurately detect the c.2T>C mutation of SLC25A13 gene, which is helpful for the diagnosis of citrin deficiency (CD).

To establish the amplification-refractory mutation system quantitative real-time PCR (ARMS-qPCR) method based on qPCR technique for detecting the c.2T>C mutation of SLC25A13 gene and validate its diagnostic performance. According to the principle of ARMS-qPCR primer design, the specific primers were designed for the conserved sequence of SLC25A13. The c.2T>C mutation ARMS-qPCR detection assay of SLC25A13 gene and the corresponding Sanger sequencing system were established through the use of the synthetic plasmids of homozygous mutation and 200 human peripheral blood specimens which were verified by Sanger sequencing as templates, and the diagnostic efficacy of the qPCR assay was validated by using nucleic acid extracted from another 200 human peripheral blood specimens and the results obtained were compared with the Sanger sequencing results as the gold standard, and the consistency of the two detection methods was analyzed. The results showed that the qPCR assay could accurately identify artificial plasmids carrying different mutations of SLC25A13 gene, and distinguish between wild type SLC25A13 gene and the c.2T>C mutation. This method was used to detect the mutation status of SLC25A13 c.2T>C in human peripheral blood, and the detection results were 100% consistent with the Sanger sequencing results. Among the 200 blood samples, 8 samples (4%) carried the c.2T>C mutation of SLC25A13 gene and 192 samples (96%) did not carry it. In conclusion, the ARMS-qPCR test established in this study can quickly, simply and accurately detect the c.2T>C mutation of SLC25A13 gene, which is helpful for the diagnosis of citrin deficiency (CD).

Hepatic fibrosis develops as a wound-healing scar in response to acute and chronic liver inflammation and can lead to cirrhosis in patients with chronic hepatitis B and C. The condition arises due to increased synthesis and reduced degradation of extracellular matrix (ECM) and is a common pathological sequela of chronic liver disease. Excessive deposition of ECM in the liver causes liver dysfunction, ascites, and eventually upper gastrointestinal bleeding as well as a series of complications. However, fibrosis can be reversed before developing into cirrhosis and has thus been the subject of extensive researches particularly at the gene level. Currently, therapeutic genes are imported into the damaged liver to delay or prevent the development of liver fibrosis by regulating the expression of exogenous genes. One technique of gene delivery uses ultrasound targeting of microbubbles combined with therapeutic genes where the time and intensity of the ultrasound can control the release process. Ultrasound irradiation of microbubbles in the vicinity of cells changes the permeability of the cell membrane by its cavitation effect and enhances gene transfection. In this paper, recent progress in the field is reviewed with emphasis on the following aspects: the types of ultrasound microbubbles, the construction of an ultrasound-mediated gene delivery system, the mechanism of ultrasound microbubble-mediated gene transfer and the application of ultrasound microbubbles in the treatment of liver fibrosis.

Artificial blood vessels, serving as crucial vascular substitutes, have been widely utilized in vascular interventional therapies and revascularization surgeries. Small-diameter artificial blood vessels (diameter < 6 mm) pose challenges for long-term implantation due to their small diameter, slow flow velocity, low blood pressure, and complex blood flow environment. Bacterial nanocellulose (BNC), a natural polymer material, enhances the regenerative and repair effects of small-diameter artificial blood vessels through composite modification and surface modification. This article reviewed the research progress in the preparation of small-diameter artificial blood vessels using BNC and discussed the advantages and potential application prospects of BNC artificial blood vessels.

Objective To construct and identify an overexpressing recombinant adenovirus vector carrying the mouse nucleotide-binding oligomerization domain-like receptor protein 6 (NLRP6) gene (NM_133 946.2). Methods The coding sequence of the mouse NLRP6 gene was synthesized, digested with enzymes, and inserted into the adenovirus vector (GL2003). The transformed cells were then cultured in strain DH-5α, transformants were selected, and after identification by polymerase chain reaction (PCR), samples were sent for sequencing and comparison. The expression of NLRP6 protein in the recombinant plasmid was validated using the Western blot. The recombinant adenovirus vector Ad-NLRP6 was obtained using the Admax packaging system, then transduced into HEK293 cells, amplified, purified, and the viral titer was determined afterwards. Results The successful integration of NLRP6 into the expression plasmid was confirmed through double enzyme digestion and DNA sequencing. Concurrent detection of the FLAG-tagged protein indicated a significant increase in the expression level of NLRP6. Recombinant adenovirus carrying NLRP6 were packaged and produced, followed by infecting HEK293 cells with the virus. The cellular expression of green fluorescent protein in Ad-NLRP6 indicated successful infection. The virus was collected, amplified, purified, and the viral titer was determined to be 4×10¹⁰ PFU/mL. Conclusion The overexpression recombinant adenovirus vector carrying the mouse NLRP6 gene has been successfully constructed, providing an effective tool for further exploring the molecular function of NLRP6.