Establishment and Characterization of a New Intrahepatic Cholangiocarcinoma Cell Line, ICC-X2.

Background: Intrahepatic cholangiocarcinoma (ICC) is an aggressive malignant tumor of the biliary tract that is prone to recurrence and metastasis and is characterized by poor sensitivity to chemotherapy and overall prognosis. For these reasons, there is an urgent need to understand its pathological mechanisms and develop effective treatments. To address this challenge, the establishment of suitable preclinical models is critical. Methods: Fresh ICC tissue samples were used for primary culture and subculture. The cell line was evaluated by cell proliferation assays, clonal formation assays, karyotype analysis, and short tandem repeat (STR) analysis. Drug resistances against oxaliplatin, paclitaxel, gemcitabine and 5-fluorouracil (5-FU) were evaluated by CCK-8 assay. Subcutaneous injection of 1 × 106 cells to three BALB/c nude mice was conducted for xenograft studies. The hematoxylin and eosin (H&E) staining was used to detect the pathological status of the cell line. The expression of biomarkers CK7, CK19, Ki-67, E-cadherin and vimentin was determined by immunocytochemistry assay. Results: A new ICC cell line named ICC-X2 was successfully established. Like ICC-X3 established using the same patient's metastatic tumor, the cell line has been continuously cultured in vitro for more than a year and has been passaged more than 100 times. ICC-X2 retained the typical biliary epithelial morphology. The population doubling time of ICC-X2 is 48 h. The cells demonstrated an abnormal nearly tetraploid karyotype. The STR analysis confirmed that ICC-X2 was highly consistent with the primary tumor tissue and not cross-contaminated by existing cell lines. ICC-X2 cells positively expressed CK7, CK19, E-cadherin, and vimentin, and the positive expression of Ki-67 in ICC-X2 cells was 40%. The ICC-X2 cells exhibited a strong clonogenic ability. The drug sensitivity test indicated that ICC-X2 was sensitive to oxaliplatin and paclitaxel, but naturally resistant to gemcitabine and 5-FU. ICC-X2 was rapidly able to form transplanted tumors in vivo after subcutaneous inoculation in nude mice. Conclusions: ICC-X2 is an excellent experimental model that can be used for studying the occurrence, development, and metastasis of ICC and investigating the mechanism of tumor drug resistance.

Quantitative assessment and influence factors of facial wrinkle situation in male construction workers in Beijing.

OBJECTIVE: To investigate current situation of facial wrinkles of male construction workers in Beijing area and to discuss the correlative factors. MATERIALS AND METHODS: A total of 149 male construction works and 63 male non-construction workers in Beijing were required to complete a questionnaire on their exposure to sunlight, dust, noise, and heat in their workplace environment. Their facial wrinkle scores were measured by VISIA Complexion Analysis System. The two-sample t test, chi-square test, and multiple linear regression were used for statistical analysis RESULTS: The exposure to sunlight, dust, noise, and heat of construction workers was significantly higher than that of non-construction workers (P < .01). The wrinkle score of construction workers between 20 and 29 years old was significantly higher than that of non-construction workers (t = 4.077, P < .01). The facial wrinkle score of construction workers(r = 0.657, P < .01) and non-construction workers (r = 0.681, P < .01) was both positively correlated with age. The wrinkle score of construction workers was related to age, sunlight, and noise(P < .01). CONCLUSION: The wrinkle score of male construction workers between 20 and 29 years old is significantly higher than that of non-construction workers in Beijing. Age, sunlight, and noise were the main influencing factors of wrinkle.

[Oridonin inhibits proliferation of Jurkat cells via the down-regulation of Brg1].

OBJECTIVE: To investigate the effect of oridonin on the human acute lymphocytic leukemia cell line Jurkat and its mechanism. METHODS: Jurkat cells were cultured in vitro and treated with various concentrations (0, 1.25, 2.5, 5, and 10 µmol/L) of oridonin for different lengths of time (24, 48, and 72 hours). The proliferation of Jurkat cells was analyzed by MTT assay. The changes in nuclear morphology were evaluated by fluorescence microscopy at 12 hours after treatment with various concentrations of oridonin. The expression levels of Brg1, P53, and C-myc were determined by semi-quantitative Western blot in Jurkat cells treated with various concentrations of oridonin for 24 hours or 5 µmol/L oridonin for various lengths of time (0, 2, 6, 12, and 24 hours). The expression levels of P53 and C-myc and proliferation of Jurkat cells were evaluated after Brg1 expression was knocked down by Brg1-specific siRNA. RESULTS: Compared with the control group, the proliferation of oridonin-treated Jurkat cells was significantly inhibited in a concentration- and time-dependent manner (P<0.05). According to the florescence microscopic analysis, oridonin treatment led to nuclear pyknosis in Jurkat cells. Compared with the control group, Jurkat cells treated with 5 µmol/L oridonin had reduced expression of Brg1 and C-myc but elevated expression of P53. Brg1 knock-down led to a significant reduction in proliferation of Jurkat cells (P<0.05), up-regulated expression of P53, and down-regulated expression of C-myc. CONCLUSIONS: Oridonin can inhibit the proliferation of Jurkat cells, probably via the Brg1 signaling pathway.

[Treatment of Facial Acne Vulgaris by Chinese Medicine Combined Western Medicine].

Objective To comparatively observe clinical efficacies of Fusidic Acid Cream (FAC) , Longzhu Ointment (LO) , and their combination of minocycline hydrochloride for treating facial acne vulgaris. Methods Totally 186 patients with acne vulgaris were randomly assigned to the FAC group (103 cases) and the LO group (83 cases). Each group was further divided into two subgroups ac- cording to the severity of acne: single treatment group and united treatment group. Patients with mild ac- ne vulgaris in the FAC group received FAC alone (39 cases) , and those with severe acne vulgaris in the FAC group received FAC and minocycline hydrochloride (64 cases). Patients with mild acne vulgaris in the LO group received LO alone (27 cases) , and those with severe acne vulgaris in the LO group received LO and minocycline hydrochloride. The therapeutic course for all was 4 weeks, with one return vis- it once per week. Grading of skin lesions was assessed by global acne grading system (GAGS). Clinical improvement was evaluated. Skin spots, red areas, and other data were statistically analyzed by VISIA skin analyzer. Results GAGS score was statistically different between before and after treatment in the FAC group and the LO group (P <0. 05). The total effective rate was 64. 1% (25)39) in single treatment group of the FAC group and 66. 7% (18/27) in single treatment group of the LO group, but with no statisti- cal difference between the two groups (Χ² =0. 09, P >0. 05). The total effective rate was 70. 3% (45/64) in united treatment group of the FAC group and 62. 5% (35/56) in united treatment group of the LO group, but with no statistical difference between the two groups (Χ² =0. 04, P >0. 05). Results of VISIA showed, compared with before treatment, statistical difference existed in red area of single treatment group of the FAC group and the LO group (P <0. 05). Statistical difference existed in ultraviolet rays, red area, sclererythrin of united treatment group of the FAC group and the LO group (P <0. 05). Conclusions FAC and LO could effectively control the inflammation of acne. LO had a rapid onset. Combined with minocy- cline hydrochloride, FAC could significantly reduce the secretion of fats, and LO could defense against ultraviolet more significantly.

Mutation detection of type II hair cortex keratin gene KRT86 in a Chinese Han family with congenital monilethrix.

BACKGROUND: Monilethrix is an autosomal dominant hair disorder characterized clinically by alopecia and follicular papules. In this study, we collected a Han monilethrix family to detect the mutations in patients and investigated the correlation between the genotype and phenotype of monilethrix. METHODS: In this study, we identified a Chinese family with monilethrix through light microscopic and scanning electron microscopic (SEM) examination. Genomic DNA from peripheral blood samples was prepared. DNA samples from controls and monilethrix patients were subject to polymerase chain reaction (PCR) amplification. Two pairs of primers were used to amplify the seventh exon of KRT86. Mutation screening of the PCR products was detected using direct sequencing. RESULTS: Light microscopic examination showed a regular alternate enlargement and narrow area. SEM examination showed that part of the cuticle of the nodules shed and disappeared gradually in the narrow area with granular protrusions on the surface similar to the erosion-like structure. Parallel longitudinal ridge and groovepattern appeared, and the ridges varied in width, like dead wood. A heterozygous transversion mutation c.1204G > A (p.E402K) in the seventh exon of KRT86 was identified in both patients. CONCLUSIONS: The mutation of extron 7 of KRT86 identified plays a major role in the pathogenesis of this pedigree with monilethrix, and is a mutation hot spot of KRT86. Further research is needed to explore the relationship between the phenotype and the mutation of the type II hair keratin gene KRT86 of monilethrix.

[Determination of vitexin in plasma by HPLC-MS/MS method and its pharmacokinetics in rats].

OBJECTIVE: To establish a HPLC-MS/MS method for the determination of vitexin in rat plasma and its pharmacokinetics. METHODS: The HPLC-MS/MS method used Capcell Pak C18 column (50 mm x 2.0 mm I. D., 5 microm). The mobile phase was methanol and water (95:5, V/V, containing 0.1% formic acid) at a flow rate of 0.2 mL/min. Electrospray ionization (ESI) in negative ion mode and multiple reaction monitoring (MRM) was used for the quantification of vitexin with a monitored transitions m/z 431-->311 for vitexin and m/z 269-->225 for internal standard (I. S., emodin). RESULTS: Linear calibration curves were obtained over the concentration range of 0.5-2000 ng/mL (r = 0.9960) with the lowest limit of quantification (LLOQ) of 0.5 ng/mL. The recovery was in the range of 76.1%-89.0%. The relative standard deviations for the intra-day and inter-day validation were less than 11%. CONCLUSION: The method is simple, accurate, fast, sensitive and suitable for the pharmacokinetic study of vitexin in rats.