Objective:To determine the expression of matrix metalloproteinase 13 (MMP13) in patients with psoriasis, and to evaluate the effect of tazarotene and narrow-band ultraviolet B (NB-UVB) on the expression of MMP13 in mice with psoriasis-like dermatitis.Methods:Lesional skin tissues and normal skin tissues were collected from 18 patients with psoriasis vulgaris and 10 healthy controls respectively, who were enrolled from General Hospital of Tianjin Medical University between May 2019 and August 2019, and serum samples were collected from all the subjects. A total of 25 specific pathogen-free (SPF) male BALB/c mice were randomly divided into control group, imiquimod group, imiquimod+NB-UVB group, imiquimod+tazarotene group and imiquimod+tazarotene+NB-UVB group. The control group received topical vaseline cream on the back once every morning; imiquimod group and imiquimod+NB-UVB group received imiquimod cream on the back once every morning; imiquimod+tazarotene group and imiquimod+tazarotene+NB-UVB group received imiquimod cream on the back once every morning, and tazarotene cream on the back once at night; imiquimod+NB-UVB group and imiquimod+tazarotene+NB-UVB group received NB-UVB irradiation on the back every other day at noon, with the dose being 300 mJ/cm 2 in the first session and increasing by 50 mJ/cm 2 in every session. The modeling lasted 7 days. After successful modeling, blood samples were obtained from the eyeballs of the mice, and skin tissues were resected from the back of the mice after being sacrificed by cervical dislocation on day 8. Changes in the epidermal thickness and pathological manifestations were observed by hematoxylin and eosin (HE) staining, protein expression of MMP13 in skin tissues was determined by immunohistochemical study, and the serum level of MMP13 was detected by enzyme-linked immunosorbent assay. Comparisons between 2 groups were performed by using two-independent-sample t test, comparisons among several groups by using one-way analysis of variance, multiple comparisons by using least significant difference- t test, and comparisons of enumeration data by using chi-square test. Results:The skin lesions of the patients with psoriasis were strongly positive for MMP13, and the MMP13 expression levels in the epidermis and serum (84.11±17.16, 13.29±3.95 μg/L, respectively) were significantly higher in the patients with psoriasis than in the healthy controls (11.98±4.08, 7.46±1.58 μg/L, respectively, both P< 0.01) . Compared with the control group (1.26±0.04 μm, 25.40±2.34, 185.76±7.22 μg/L, respectively) , a significant increase was observed in the epidermis thickness (7.93±0.59 μm, P< 0.01) , as well as MMP13 levels in the epidermis and serum in the imiquimod group (147.14±5.53, 215.98±15.17 μg/L, respectively, both P< 0.01) . Compared with the imiquimod group, the imiquimod+tazarotene group, imiquimod+NB-UVB group, and imiquimod+tazarotene+NB-UVB group all showed significantly decreased epidermal thickness (3.56±0.37 μm, 3.83±0.39 μm, 2.14±0.34 μm, respectively, all P< 0.05) , MMP13 levels in the epidermis (120.42±3.23, 91.08±0.46, 71.12±7.11, respectively, all P< 0.05) and serum (197.39±3.92 μg/L, 196.13±11.76 μg/L, 183.21±14.99 μg/L, respectively, all P< 0.05) . Conclusions:MMP13 protein expression markedly increased in the skin lesions and sera of patients with psoriasis, and decreased in skin lesions and sera of mice with psoriasis-like dermatitis after the treatment with tazarotene and NB-UVB. MMP13 may be involved in the development of psoriasis, and tazarotene and NB-UVB may inhibit the development of psoriasis by reducing the expression of MMP13.

Objective To investigate the angiogenesis effects of Salvia miltiorrhiza in heterotopically grafted frozen-thawed mouse ovaries. Methods The ovaries thawed after cryopreservation were xenografted into the donated kidney capsules of 8- to 12-week adult male mice. The mice were divided into two groups, saline and Salvia miltiorrhiza groups, the mice either in the saline or in Salvia miltiorrhiza groups were administered I.p. Daily either saline(0.5ml) or Salvia miltiorrhiza(0.5g)respectively, from the day prior transplantation. The two groups were sacrificed 1 day,2 days and 7 days after transplantation respectively, the grafts from thawed,1 day,2 days,7 days were removed for follicle counting, immunohistochemical studying and detecting of the mRNA expression of vascular endothelial growth factor(VEGF). Results The number of follicles and survival rates in grafts after transplantation of Salvia miltiorrhiza group were more than that of saline group (P<0.05);the expression of VEGF increased after transplantation,peaked on day 7,there was no difference between the two groups (P>0.05);the apoptosis index of Salvia miltiorrhiza group was less than that of saline group (P<0.05), the mRNA expression of VEGF188 and VEGF164 of Salvia miltiorrhiza group was more than that of saline group on 48 hours after transplantation(P<0.05). Conclusion Salvia miltiorrhiza may provide benefits for folliculogensis and decreasing the apoptosis index of follicles. Nevertheless,a increase in the VEGF188 and VEGF164 isoform in the Salvia miltiorrhiza group may suggest the positive effect of exogenous Salvia miltiorrhiza therapy in the early stage of angiogenesis.

Background The incidence of dry eye is increasing among young adults because of wide usage of video display terminal.But the early diagnosis of dry eye still presents challenge to medical practitioners.The accurate diagnosis and treatment of the dry eye,therefore,is a topic of high interest to researchers.Previous examination outcome of dry eye is interferred primarily due to invasive procedure.It is very important to search an examination approach.Objective This study was to use Keratograph 5M,a non-invasive ocular surface analyzer to evaluate the influence of watching video display terminal on ocular surface and tear film.Methods Eighty-one eyes of 81 health volunteers among 18-30 years were enrolled in Affiliated Eye Hospital of Nanchang University from March 1,2015 to November 10,2015 under the informed consent,including 39 males and 42 females.The subjects watched the computer for continuously 3 hours under the nature light,and ocular surface related examinations were performed and compared before and after video display terminal exposure,including non-invasive tear film break-up time (NITBUT),tear meniscus height,conjunctival hyperemia scoring,limbal congestion scoring,corneal fluorescein staining scoring,meibomian gland imaging and lipid layer analysis.Results The number of eyes with visual fatigue,dryness,pain,blurring and conjunctival congestion was significantly increased after 3-hour video display terminal exposure in comparison with before (all at P＜0.01).The initial NITBUT and mean NITBUT were (6.086± 3.701) s and (9.103 ± 4.680) s,and tear meniscus height was (0.190 ± 0.032) mm after trail,which were significantly lower than (11.445 ±4.964) s,(14.626 ±4.467) s and (0.212 ±0.040) mm of before trail,respectively;The conjunctical hyperemia scoring and limbal congestion scoring were 0.869 ±0.311 and 0.572 ±0.276 after trial,which were significantly higher than 0.780 ± 0.306 and 0.509 ± 0.266 before trail,showing significant differences before and after exposure of video display terminal (all at P＜0.01).The intraocular pressure and the eye number of different scores of corneal fluorescence staining,abnormal meibomian gland and different morphological lipid layer of tear were unchanged before and after exposure of video display terminal.Conclusions Long-term exposure of video display terminal results in significant and temporary adverse influence on tear film and ocular surface.Keratograph 5M non-invasive ocular sudace analyzer can objectively assess overall ocular surface conditions.

Objective To evaluate the change of left atrial function after mitral valvuloplasty in patients with mitral valve prolapse by measuring left atrial strain using two-dimensional speckle tracking echocardiography(2D-STE).Methods Study population consisted of 25 patients with mitral valve prolapse and 25 healthy subjects.High frame rate two-dimensional images were recorded from the apical four chamber view and two chamber view.Images of patients with mitral valve prolapse were respectively recorded at pre-operation,3 days and 3 months post-operation.Left atrial longitudinal strain was measured in two views using two-dimensional strain soft ware.Results ①Compared with the control group,global peak atrial longitudinal strain (PALS),peak atrial contraction strain (PACS) and atrial longitudinal strain during early diastole(ALSED) decreased (P ＜0.01) in patients with mitral valve prolapse,but global timeto-peak of peak atrial longitudinal strain(TPLS) increased(P ＜0.01).Regurgitant jet area/left atrium area (RJA/LAA) in patients with mitral valve prolapse correlated negatively with global PALS(r =-0.620,P ＜ 0.01).② Compared with preoperative group,global PALS,PACS,ALSED and TPLS decreased significantly at 3 days after surgery(P ＜0.01).Global PALS,ALSED(P ＜0.01) and PACS(P ＜0.05)increased at 3 months after surgery,but global TPLS did not differ significantly.Conclusions 2D-STE could accurately evaluate the change of left atrial function after mitral valvuloplasty.

Objective To evaluate the application value of real-time three-dimensional echocardiography (RT-3D TEE) in nonvalvular atrial fibrillation patients after transcatheter left atrial appendage closure (LAAC) with the Amplatzer Cardiac Plug (ACP).Methods The two-dimensional transesophageal echocardiography (2D TEE) and RT-3D TEE were performed in selected patients to measure the diameter of left atrial appendage ostium and landing zone,to observe left atrial appendage morphology and lobular distribution.These were also performed to guide the whole process of transcatheter LAAC with ACP,which included the atrial septal puncture,sheathing canal cruise,occluder device implantation,and verifying the stability of occluder device and releasing the device.It involved observing ACP occluder morphology,location,stability,surrounding residual shunt,whether influencing the mitral valve and left superior pulmonary vein flow,and pericardial effusion.Results A total of 15 patients (100％) successfully underwent LAAC with the ACP.The maximum diameter of left atrial appendage ostium by 2D-TEE measurement during operation was (20.5 ± 2.9)mm,located at 135°.The diameter of left atrial appendage landing zone was (17.1 ± 2.8) mm,located at 45°;(18.0 ± 4.0) mm,located at 90°;(22.1 ± 4.7)mm,located at 135°,respectively.The left atrial appendage morphology:2 had one leaf and 13 had two leaves or more,including 2 cases of bifoliate short neck shape.In 15 patients,6 cases of cauliflower type,2 cases of wind sock type,3 cases of chicken wing type,2 cases of cactus type and 2 cases of complex type.The proximal left atrial appendage morphology:3 cases of boot type,2 cases of wide mouth type,6 cases of narrow mouth type,2 cases of straight tube type,and 2 cases of bifoliate short neck type.There was no obvious residual shunt in all the patients at immediately post-operation.Conclusions In the transcatheter LAAC with the ACP,RT-3D TEE has important application value in the preoperative selection of patients,the choice of appropriate occluder,guidance of full-process monitoring during operation,the postoperative effect evaluation and so on.

This study was aimed to establish rat bladder tumor animal models to investigate the in viva antitumor effect of polyanhydride-pirarubicin (PAD-THP), a long-lasting anti-cancer implant, in the bladder tumor of animal models. The model of bladder cancer was set up with N-butly-N-(4 hydroxybutyl) nitrosamine (BBN) feeding into rats. The PAD-THP long-acting anti-cancer implants containing the drugs and the same dose of the THP naked drug were placed under the bladder mucosa of bladder tumor model in vivo. The pirarubicin plasma concentration was measured with high performance liquid chromatography (HPLC) detection in vivo. The effective drug concentration and lasting period were observed and compared in the animal bodies. The tumor sizes were measured before and after the treatment. The in viva antitumor effects were analyzed and compared. The results showed that more significant antitumor effect of PAD-THP implants on the local drug release characteristics were presented compared with that of the same dose of THP bare drug group and there were significant differences (P<0. 05) between the two methods. All the results indicated that the PAD-THP anti-cancer implants in the postoperative local treatment of bladder tumors would show prosperous in the future for clinical application.
Animals ; Antineoplastic Agents ; administration & dosage ; Butylhydroxybutylnitrosamine ; Delayed-Action Preparations ; administration & dosage ; Disease Models, Animal ; Doxorubicin ; administration & dosage ; analogs & derivatives ; Female ; Implants, Experimental ; Polyanhydrides ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Urinary Bladder Neoplasms ; chemically induced ; drug therapy ; pathology

OBJECTIVETo explore the feasibility of IGF2 imprinting system in target gene therapy for tumors.

METHODSThe mouse H19 enhancer, DMD and promoter H19 were amplified by PCR from mouse genomic DNA and then cloned into the plasmid pDC312. The EGFP and DT-A fragments were amplified by PCR and cloned into the recombinant plasmid, and then the shuttle plasmid were transfected into HEK293 cells together with the adenoviral vector Ad5, namely, Ad-EGFP and Ad-DT-A. Adenovirus hexon gene expression was applied to confirm the presence of adenovirus infections. The effect of the IGF2 imprinting system was tested by fluorescence microscopy. RT-PCR and Western blotting after transfection of the recombinant adenoviral vectors into cancer cells were used to show loss of IGF2 imprinting (LOI) and maintenance of IGF2 imprinting (MOI), respectively. The anti-tumor effect was assessed by MTT and flow cytometry after the HCT-8 (LOI). Human breast cancer cell line MCF-7 (MOI) and human normal gastric epithelial GES-1 (MOI) cell line were transfected with Ad-DT-A in vitro. The anti-tumor effect was detected by injecting the Ad-DT-A in nude mice carrying HCT-8 tumors.

RESULTSThe expression of EGFP protein, DT-A mRNA and DT-A protein were seen to be positive only in the HCT-8 tumor cell line. Infection with Ad-DT-A resulted in obviously growth inhibition in HCT-8 cells (75.4 ± 6.4)% compared with that in the control group, and increased the percentage of apoptosis in the HCT-8 cells (20.8 ± 5.9)%. The anti-tumor effect was further confirmed by injecting the recombinant adenoviruses in HCT-8 tumor-bearing nude mice, and the results showed that the Ad-DT-A inhibited the tumor growth, with on inhibition rate of 36.4%.

CONCLUSIONSThe recombinant adenoviruses carrying IGF2 imprinting system and DT-A gene have been successfully constructed, while Ad-DT-A can effectively kill the tumor cells showing loss of IGF2 imprinting. It might play an important role in future target gene therapy against malignant tumors based on loss of IGF2 imprinting.

Adenoviridae ; genetics ; Animals ; Apoptosis ; Breast Neoplasms ; genetics ; pathology ; Colonic Neoplasms ; genetics ; pathology ; therapy ; Diphtheria Toxin ; biosynthesis ; genetics ; Female ; Genetic Therapy ; methods ; Genetic Vectors ; Genomic Imprinting ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Humans ; Insulin-Like Growth Factor II ; genetics ; metabolism ; MCF-7 Cells ; Mice ; Mice, Nude ; Neoplasm Transplantation ; Peptide Fragments ; biosynthesis ; genetics ; Plasmids ; RNA, Messenger ; metabolism ; Random Allocation ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Transfection

Objective To identify the differences inneuropsychological characteristics between amnestic(AMCI)and vascular mild cognitive impairment(VMCI).Methods Totally 297 old community residents with mild cognitive impairment(MCI)were divided into amnestic MCI(AMCI)and vascular MCI(VMCI)subgroup from Guangzhou MCI prevalence survey.The elderly with MCI were interviewed and tested with the Chinese version of Montreal Cognitive Assessment(MoCA),the Mini-Mental state examination(MMSE),Auditory Verbal Learning Test(AVLT),the Clinical Dementia Rating scale(CDR),Functional Activity Questionnaire(FAQ),the Modified Hachinski Ischemic Scale(M-HIS),Center for Epidemiologic Studies(CES-DC)to evaluate neuropsychological characteristics.Results AMCI versus VMCI group showed that the total scores of MoCA were(9.63±5.17 vs.9.98±6.02),total scores of MMSE were(16.90±4.84 vs.16.90±6.19),AVLT immediate memory was(2.35±1.39 vs.2.91±1.84),AVLT delayed recall was(2.23±2.09 vs.2.47±2.20),AVLT delayed recognition was(7.33±3.98 vs.6.85±4.02)and total scores of CDR(0.5 vs.0.5),with no differences between the 2 groups(all P>0.05).Based on MoCA survey,AMCI versus VMCI group showed statistically significant differences(all P<0.05)in parameters of visual space and execution(0.71±1.02 vs.0.92±1.26),language function(0.34±0.56 vs.0.50±0.80)and abstract thinking(0.25±0.49 vs.0.15±0.43),but based on MMSE survey,no difference was found in the various cognitive domains between the two groups.The AMCI versus VMCI group showed statistically significant differences(all P<0.05)in parameters of CES-DC scale(1.75±4.27 vs.2.76±6.72),FAQ scale(4.42±4.66 vs.8.71±7.03),M-HIS scale(0.40±0.64 vs.7.59±3.53).Conclusions There is no significant difference in general cognitive impairment between AMCI and VMCI,but the visual space and execution,language function are more impaired in AMCI than VMCI,and the abstract thinking,social function are more impaired with more depressive symptoms in VMCI than in AMCI.

Objective:To investigate the biological characteristics of proliferation, apoptosis, migration and invasion of radiation-induced polyploid cervical cancer HeLa cells, and to analyze the potential facilitation of polyploid HeLa cells in cervical cancer recurrence after radiotherapy.Methods:HeLa cells were irradiated by 6 MV-X ray with 7 Gy and 14 Gy, the cells were cultured until the third day, and then they were recorded as 7 Gy group and 14 Gy group respectively. The unirradiated HeLa cells were recognized as the control group. The cell morphology was checked under optical microscope. Flow cytometry was used to determine cell ploidy. MTT assay was applied to detect cell proliferation. Flow cytometry by AnnexinⅤ-FITC/PI double labeling was used to detect apoptosis. The ability of migration and invasion was detected by Transwell assay. The expression levels of STAT3 signal pathway-related proteins were analyzed by Western blotting.Results:Compared with the control group, the volume of HeLa cells in 7 Gy group and 14 Gy group increased significantly. The percentages of polyploid HeLa cell subsets in the control group, 7 Gy group and 14 Gy group were (6.33±1.26) %, (21.13±0.50) % and (46.07±1.68) % respectively, with a statistically significant difference ( F=780.47, P<0.001) . The absorbance values in the control group, 7 Gy group and 14 Gy group of polyploidy HeLa cells were 0.21±0.01, 0.23±0.02, 0.16±0.01 at 24 h, 0.37±0.03, 0.38±0.06, 0.21±0.00 at 48 h, 0.66±0.02, 0.55±0.01, 0.28±0.01 at 72 h, and there were statistically significant differences ( F=31.62, P=0.001; F=20.10, P=0.002; F=708.52, P<0.001) . Further pairwise comparison showed that the proliferation abilities of polyploidy HeLa cells of the 14 Gy group at 24, 48 and 72 h were significantly lower than those of the control group and the 7 Gy group (all P<0.05) . The proportions of apoptotic cell subset in the control group, 7 Gy group and 14 Gy group were (3.67±1.16) %, (3.07±0.81) %, (3.83±0.91) %, the proportions of early apoptotic subset were (2.33±0.35) %, (2.13±0.61) %, (2.23±0.32) %, and the proportions of late apoptotic subset were (1.33±0.81) %, (0.93±0.31) %, (1.60±0.60) % respectively. There were no statistically significant differences ( F=0.52, P=0.620; F=0.15, P=0.864; F=0.92, P=0.450) . The migrated numbers of cells in the control group, 7 Gy group and 14 Gy group were 297.40±26.53, 121.33±15.16, 18.40±4.79, and the invaded numbers were 195.67±20.26, 63.60±6.91, 9.47±3.23 respectively, with statistically significant differences ( F=647.28, P<0.001; F=213.94, P<0.001) . Compared with the control group, the migration and invasion abilities of polyploid HeLa cells in the 7 Gy and the 14 Gy groups were significantly decreased, and the migration and invasion abilities of polyploid HeLa cells in the 14 Gy group were significantly lower than those in the 7 Gy group (all P<0.001) . The expression levels of P-STAT3 (Tyr 705) and Bcl-2 in radiation-induced polyploidy HeLa cells were higher than those in the control group, and the expression levels were further increased with the increase of radiation dose. Compared with the control group, the expression levels of Survivin and Mcl-1 in polyploid HeLa cells in the 14 Gy group were up-regulated (both P<0.05) . There was no significant difference in Bcl-xL expression among the three groups ( F=0.52, P=0.618) . Conclusion:The proliferation, migration and invasion abilities of polyploid HeLa cells are reduced by radiation, and the proportion of apoptotic subset is not significantly changed, but the activation of STAT3 signaling pathway is accompanied by up-regulation of downstream anti-apoptotic related proteins, which is favorable for the polyploid tumor cells to be the potential risk factor of recurrent cervical cancer after radiotherapy.

During the screening of a traditional Chinese folk herb library against HepG2 and Hep3B cell lines, the EtOAc extract from the Tibetan medicine, Caragana tibetica (CT-EtOAc) exhibited potential anti-hepatocellular carcinoma (anti-HCC) activity. HPLC-based activity profiling was performed for targeted identification of anti-HCC activity from CT-EtOAc by MS-directed purification method. CT-EtOAc was separated by time-based fractionation for further anti-HCC bioassay by a semipreparative HPLC column (150 mm × 10 mm i.d., 5 μm) with a single injection of 5 mg. Bioassay-guided and ESIMS-directed large scale purification was performed with a single injection of 400 mg of CT-EtOAc by peak-based fractionation. A 1.4-mm heavy wall micro NMR tube with z-gradient was used to measure one and two dimensional NMR spectra for the minor or trace amounts of components of the extract. Two active compounds could be elucidated as naringenin chalcone (CT-1) and 3-hydroxy-8, 9-dimethoxypterocarpan (CT-2) relevant to anti-HCC effects for the EtOAc extract of C. tibetica rapidly and unambiguously by this protocol.
Acetates ; pharmacology ; Antineoplastic Agents ; chemistry ; pharmacology ; Caragana ; chemistry ; Carcinoma, Hepatocellular ; drug therapy ; Cell Line, Tumor ; Chalcones ; pharmacology ; Chromatography, High Pressure Liquid ; Hep G2 Cells ; Humans ; Liver Neoplasms ; drug therapy ; Medicine, Tibetan Traditional ; Plant Extracts ; chemistry ; pharmacology ; Plant Roots ; chemistry