Purpose To investigate the diagnostic value of p16/Ki-67 dual stain cytology for detection of cervical precancerous lesions as a novel option for cervical lesions screening.Methods A total of 295 cases diagnosed as atypical squamous cells of undetermined significance (ASC-US) and low-grade squamous intraepithelial lesion (LSIL) from thinprep cytologic test (TCT) were selected.Double staining of p16/Ki-67 cytology,vaginal biopsy,biopsy and pathological examination were applicated,p16/Ki-67 dual stain cytology was compared with that of biopsy and pathological examination.At the same time,The sensitivity and specificity of p16/Ki-67 dual stain cytology between ASC-US and LSIL was analyzed.Results The positive rate of p16/Ki-67 dual stain cytology were 37.42％ and 36.36％ in ASC-US and LSIL,respectively.The positive rate of cervical intraepithelial neoplasia 2/3 (CIN2/3) were 25.77％ and 25.76％.The sensitivity and specificity of the p16/Ki-67 test for detecting CIN2/3 was 83.33％ and 78.51％.The sensitivity and the specificity of the p16/Ki-67 test for detecting CIN2/3 was 85.30％ and 80.61％ in LSIL group.Conclusion p16/Ki-67 double stain cytology detection can improve the sensitivity of CIN2/3 and the specificity of human papilloma virus (HPV).p16/Ki-67 double stain detection can effectively triage the high grade cervical lesions in TCT and improve the accuracy of cervical cancer screening.

Objective To investigate the effect of DRAM1 on the acute ischemic injury of H9C2 cardiomyocytes. Methods H9C2 cardiomyocytes were treated with Oxygen-glucose deprivation (OGD) after DRAM1 adenovirus transfection. MTT assay was performed to detect cell viability and Annex V/PI staining was used to analyze the cell apoptosis. Western blot was performed to measure the expression of P62. Results DRAM1 overexpression increased the H9C2 cardiomycytes viability after OGD treatment for 12 hours. DRAM1 overexpression was attenuated, while siRNA-AD-DRAM1 exacerbated the apoptosis rate of H9C2 cardiomycytes after OGD treatment for 12 hours by Annex V/PI staining. P62 protein expression was increased in the H9C2 cardiomycytes after OGD treatment for 12 hours, which was reversed by DRAM1 overexpression. Conclusion DRAM1 may protect H9C2 cardiomycytes against OGD injury due to the improvement of the autophagy flux.

Objective To observe the expression of ITF in colon of mice infected by mouse cytomegalovirus (MCMV) and the in-volvement of ganciclovir(GCV) .Methods Forty-eight BALB/c young mice were randomly divided into blank group ,virus group and GCV group ,each with 8 mice .Mice in virus group and GCV group received injection of 100 μL MCMV virus suspension (TCID50105 .31 /mL) ,and GCV group was given intraperitoneal injection of GCV once a day at the dose of 50 mg/kg from day 0 (24 hours after vaccination of virus ) ,for 14 days .At the same time the virus group and blank group were given the same dose of normal saline as controls .Murine cytomegalovirus loads in livers and colons were measured by PCR .The expression levels of ITF in mRNA in colon were detected by RT-PCR .Results After MCMV injection ,mice in virus group manifested aggressively apparent poor appetite ,less activity ,fur laxly ,unresponsiveness to stimuli ,growth retardation ,body weight not increased .All liver and colon tissue MCMV-DNA PCR electrophoresis of virus group had positive strip ,while the blank group did not .GCV group also showed less bright positive strip when compared with the virus group .Expression level of ITF mRNA was significant higher in GCV group than virus group ,there was statistically significant difference(P<0 .05) .Expression of ITF mRNA in virus group were higher than that in blank group ,there was statistical difference(P<0 .05) .Conclusion ITF is regarded as a fast reaction peptide in the course of mucosa impairments ,so ITF plays a protective role in delayed healing process after acute MCMV infection .

OBJECTIVE: To explore the effect of the composition ratio on substitution of sulfate group in sulfated exopolysaccharide (EPS) from and how sulfate modification affects the anti-tumor activity of EPS. METHODS: We used a chlorosulfonic acid-pyridine method to modify EPS and analyzed the effect of esterification ratio on the degree of sulfate substitution using barium chloride turbidimetry. The sulfate groups binding with EPS were analyzed with infrared spectrum analysis. CCK-8 assay was used to evaluate the inhibitory effect of EPS sulfate (SEPS) on the proliferation of human colon cancer HCT 116 cells, and annexin V-FITC/PI double staining was used to assess the pro-apoptotic effect of SEPS in the cells. RESULTS: The esterifying agent and EPS at the composition ratios of 1:1 and 2:1 resulted in sulfate substitution of 0.98% (SEPS-1) and 1.18% (SEPS-2), respectively, and the substitution was improved by increasing the ratio of the esterifying agent ( < 0.05). Infrared spectrum analysis showed that the S=O stretching vibration absorption peak of -OSO appeared near 1249 cm, indicating that the sulfate group combined with EPS to form sulfate. CCK-8 assay showed that SEPS-1 produced stronger inhibitory effects on the proliferation of HCT 116 cells than EPS within the concentration range of 0.02-0.10 mg/L ( < 0.05). At the concentrations of 0.04-0.08 mg/L, SEPS-2 showed a lower anti-tumor activity than SEPS-1 ( < 0.05). SEPS-1 also showed stronger pro-apoptotic effect than EPS, and as its concentration increased, SEPS-1 dose-dependently increased the ratio of early apoptotic cells and necrotic cells; the cells treated with 0.06, 0.08 and 0.10 mg/mL SEPS-1 showed early apoptotic rates of 6.38%, 11.8% and 12.5%, and late apoptotic and necrotic rates of 5.26%, 8.04% and 6.80%, respectively. CONCLUSIONS The composition ratio of the esterifying agent has a direct impact on the degree of substitution of EPS, which can be improved by increasing the ratio of the esterifying agent. Sulfate modification of EPS can enhance its antitumor activity, which, however, is not directly related with the degree of substitution.

【Objective】 To research the effect of the Fc, Fab and F(ab′)2 fragments of immunoglobulin G, the main components of Human Immunoglobulin(pH4) for Intravenous Injection(IVIG), on the phagocytic function of macrophages derived from THP-1 cells. 【Methods】 First of all, IVIG was digested with papain and pepsin to obtain Fc, Fab and F(ab′)2, and these components were then identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Afterwards, propylene glycol monomethyl ether acetate (PMA) was used to induce THP-1 cells to differentiate into M0 macrophages. Finally, the sensitized erythrocytes were labeled with carboxy fluorescein succinimidyl ester (CFSE), and the effect of the above components on the phagocytic ability of M0 macrophages to engulf sensitized erythrocytes was detected by flow cytometry. 【Results】 The identification results of SDS-PAGE showed that the prepared IgG fragments met the requirements of subsequent experiments. Flow cytometry performs showed that the phagocytosis model of M0 macrophages had been successfully established. When the concentration of Fc increased from 0.1μg/ mL to 10μg/ mL, the phagocytosis rate of erythrocytes sensitized by M0 macrophages decreased from (24.21±0.58) % to (12.27±0.19) %. When the concentration of IVIG protein increased from 0.1 μg/ml to 10 μg/ml, the phagocytosis rate decreased from (20.57±0.39) % to (0.20±0.03) %. Meanwhile, at the same protein concentration (10 μg/ml), the inhibitory effect of Fc on phagocytosis was only half that of IVIG. In addition, Fab, F(ab′)2, and human serum albumin could not inhibit phagocytosis of M0 macrophages. 【Conclusion】 IVIG can effectively inhibit the phagocytosis of THP-1 derived M0 macrophages, which is mainly dependent on the Fc, but not related to the Fab of IgG and F (ab′)2.

OBJECTIVETo investigate the protective effect of monoside against triptolide-induced liver injury and explore its molecular mechanism.

METHODSBALB/C mice treated with gastric lavage with triptolide and monoside, either alone or in combination, were examined for changes of hepatic biochemical parameters using the serological method. The growth inhibition rate of HepG2 cells treated with triptolide or monoside or both was assessed with MTT assay, and the cell morphological changes were observed using laser confocal microscopy; the expressions of the target proteins in the antioxidative stress pathway were detected using flow cytometry and Western blotting.

RESULTSIn BALB/C mice, gastric lavage of triptolide induced obvious hepatic damage. In HepG2 cells, treatment with triptolide significantly inhibited the cell growth, resulting in a cell viability as low as 72.83% at 24 h; triptolide also induced obvious cell apoptosis and cell nucleus deformation, causing an apoptosis rate of 43.1% in the cells at 24 h. Triptolide significantly reduced the expressions of Nrf2 and HO-1 proteins related with the oxidative stress pathway. Combined treatment with morroniside obviously reversed these changes, resulting in significantly decreased hepatic biochemical parameters and the liver index in BALB/C mice and in significantly lowered cell apoptosis rate, improved cell morphology, and increased Nrf2 and HO-1 protein expressions in HepG2 cells.

CONCLUSIONSMonoside protects against triptolide-induced liver injury possibly by relieving oxidative stress.

OBJECTIVE: To investigate the protective effect of arctiin with anti-inflammatory bioactivity against triptolide-induced nephrotoxicity in rats and explore the underlying mechanism. METHODS: Forty SD rats were divided into 4 groups for gastric lavage of normal saline, arctiin (500 mg/kg), triptolide (500 μg/kg), or both arctiin (500 mg/kg) and triptolide (500 μg/kg). Blood samples were collected for analysis of biochemical renal parameters, and the renal tissues were harvested for determining the kidney index and for pathological evaluation with HE staining. In the RESULTS: In SD rats, arctiin significantly antagonized triptolide-induced elevation of BUN, Scr and kidney index ( CONCLUSIONS Arctiin can protect the kidney from triptolide-induced damages in rats possibly through the anti-inflammatory pathway.
Animals ; Anti-Inflammatory Agents ; Diterpenes/toxicity* ; Epoxy Compounds/toxicity* ; Furans ; Glucosides ; Kidney/drug effects* ; Phenanthrenes/toxicity* ; Rats ; Rats, Sprague-Dawley

Overexpression of exogenous lineage-determining factors succeeds in directly reprogramming fibroblasts to various cell types. Several studies have reported reprogramming of fibroblasts into induced cardiac progenitor cells (iCPCs). CRISPR/Cas9-mediated gene activation is a potential approach for cellular reprogramming due to its high precision and multiplexing capacity. Here we show lineage reprogramming to iCPCs through a dead Cas9 (dCas9)-based transcription activation system. Targeted and robust activation of endogenous cardiac factors, including GATA4, HAND2, MEF2C and TBX5 (G, H, M and T; GHMT), can reprogram human fibroblasts toward iCPCs. The iCPCs show potentials to differentiate into cardiomyocytes, smooth muscle cells and endothelial cells . Addition of MEIS1 to GHMT induces cell cycle arrest in G2/M and facilitates cardiac reprogramming. Lineage reprogramming of human fibroblasts into iCPCs provides a promising cellular resource for disease modeling, drug discovery and individualized cardiac cell therapy.