Objective To investigate the effects of low molecular heparin(LWMH)on cytokines(TNF-?,IL-1? and IL-10)in blood plasma and bronchoalveolar lavage fluid of invasive pulmonary aspergillosis(IPA)mice.Methods The neutropenic IPA mouse models were established by administration of cyclophosphamide for immunologic function inhibition and intranasally challenge with Aspergillus fumigatus conidia(1?106 conidia/mouse).One hundred and twenty mice were randomly divided into 4 groups:normal control,IPA model,normal saline+LWMH and IPA+LWMH group.Normal saline+LWMH group and IPA+LWMH group received LWMH(subcutaneous injection,1 000 IU/kg,qd?2 d).Normal control and IPA model group received normal saline instedad of LWMH.At 4,8,12,24 and 48 h after inoculation,six mice were randomly taken from each group to be sacrificed.ELISA method was used to determine the concentrations of TNF-?,IL-1? and IL-10 in blood plasma and BALF.Results TNF-?,IL-1? and IL-10 in blood plasma and BALF increased significantly several hours after inoculation of conidia in IPA model and IPA+LWMH group.There were significant higher concentrations of TNF-? and IL-1? in blood plasma and BALF in IPA+LWMH group than in IPA model group(P

In order to provide a scientific basis for the establishment of a Daphnes Cortex medicinal material fungus library and the screening of endophytic fungi that promote the growth of Daphnes Cortex and increase the content of daphnetin, we used Illumina high-throughput testing technology to analyze 9 Daphnes Cortex samples from Gansu and Shanxi provinces. A total of 632 766 valid sequences were obtained, including 348 OTUs, 4 phyla, 20 classes, 48 orders, 108 families, 154 genera, and 208 species. The sum of the first 3 fungal genera account for more than 65% of the total abundance, with the highest reaching 98.4%. Alternaria and Phoma are the main genuses of Daphne giraldii Nitsche, and Altemaria is the dominant genus. The endophytic fungi community of Daphnes Cortex is rich in diversity, and the order of fungal diversity in different producing areas is: Gangu County > Wutai County > Tanchang County.

OBJECTIVETo observe the effect of Tanshinone II A on the expression of epidermal growth facter (EGF) and epidermal growth facter recepter (EGFR) in human hepatocellular carcinoma cell line SMMC-7721.

METHODSThe human hepatocellular carcinoma SMMC-7721 cells cultured in vitro was treated with different concentrations of Tanshinone II A. The proliferation of the cells was measured by MTT assay, and the apoptosis of the cells was investigated by flow cytometry and cytochemical staining with Hoechst 33342. The expression of EGF and EGFR was detected by immunocytochemistry method. The levels of EGF in medium were measured by radioimmunoassay.

RESULTTanshinone II A inhibited the growth of SMMC-7721 cells remarkably in a dose-dependent manner. The inhibitory rate reached the peak (72.5%) after 0.5 microg/ml Tanshinone II A was used for 48 h, which was significantly higher than that in the controls (P<0.05). FCM analysis showed that when SMMC-7721 cells were treated with 0.5 microg/ml Tanshinone II A, the apoptosis rates for 24 h, 48 h and 72 h were (4.06+/-0.27)%, (7.58+/-0.56)% and (5.23+/-0.13)%, respectively which were markedly higher than those in the controls (all P<0.01). SMMC-7721 cell apoptosis with cell shrinkage, nuclear chromatin concentration and fragmentation as well as the formation of apoptotic bodies were observed by cytochemical staining when treated with Tanshinone II A. The immunocytochemistry showed that the expressions of EGF and EGFR were down regulated while the concentration of Tanshinone II A was increasing. The high expression rates for EGF and EGFR were 10%, 20%, respectively, and the gray scale was 181.52+/-1.63, 179.37+/-1.59, which were markedly higher than those in the controls (all P<0.05). The levels of EGF in medium measured by radioimmunoassay were decreased significantly after Tanshinone II A treatment.

CONCLUSIONTanshinone II A can inhibit cell proliferation and induce apoptosis in hepatocellular carcinoma cell line SMMC-7721, which may be related to the down-regulation of EGF and EGFR protein expression.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Diterpenes, Abietane ; Down-Regulation ; drug effects ; Epidermal Growth Factor ; genetics ; metabolism ; Humans ; Liver Neoplasms ; metabolism ; pathology ; Phenanthrenes ; pharmacology ; Receptor, Epidermal Growth Factor ; genetics ; metabolism

OBJECTIVETo detect the self-assembly morphology of sodium hyaluronate injection on mica using atomic force microscopy(AFM).

METHODSAtomic force microscopy with nanometer resolution was used to observe the self-assembly morphology of different concentrations of sodium hyaluronate injection on mica at room temperature.

RESULTSThe self-assembly morphology of 0.001, 0.01, and 0.1 mg/ml sodium hyaluronate injection on mica featured piebald, reticular and dendritic structures, respectively. At 1 and 5 mg/ml, sodium hyaluronate injection displayed bacilliform and spherical structures on mica, respectively; the diameter and height of the particles of 5 mg/ml sodium hyaluronate was 197.97±78.48 nm and 30.79±18.67 nm, significantly greater than those of 0.1 mg/ml sodium hyaluronate injection (49.52±11.93 nm and 5.37±1.59 nm, respectively, P<0.05).

CONCLUSIONThe self-assembly morphology of sodium hyaluronate injection on mica varies with its concentration. The piebald and reticular structure may facilitate the function of sodium hyaluronate, and the dendritic feature resembles the representative model of diffusion-limited aggregation (DLA).

Aluminum Silicates ; chemistry ; Hyaluronic Acid ; administration & dosage ; chemical synthesis ; chemistry ; Microscopy, Atomic Force ; Nanostructures ; Surface Properties

Objective To investigate the induction of microRNA (microRNA-106a,miR-106a)on the peritoneal metastasis of human gastric cancer cell BGC-823 by regulating matrix metalloproteinase inhibitor 2 (tissue inhibitor of metalloproteinases 2,TIMP2).Methods Human gastric cancer cell line BGC-823 was cultured to the logarithmic growth phase. The cells were divided into three groups: BGC-823, BGC-823/anti-miR-106a (antagomir)and BGC-823/negative control.Real-time PCR was used to identify the effect of antagomir.Transwell assay was used to detect the cell migratory and invasive abilities of these three groups in vitro .With small incision, the cells were injected into the abdominal cavity of nude mice to prepare a xenograft model.The animals were divided into two groups:miR-antagomir and miR-NC.The tumor growth in the nude mice was generally observed and estimated.Immunohistochemistry and Western blot methods were used to detect the expression of metastasis-associated protein TIMP2 on the several abdominal organs.Results The expression level of miR-106a was down- regulated in BGC-823/anti-miR-106a group,with the fold change of 0.05±0.01,which was significantly different from that in NC group (t=-18.001,P＜0.001).In vitro exogenously silencing of miR-106a gene,the numbers of invasive and migratory cells in BGC-823/anti-miR-106a group were both significantly lower than those in BGC-823 and BGC-823/negative control groups (P＜0.001).In vivo xenograft model showed that the down-regulation of miR-106a weakened the peritoneal metastasis ability of BGC-823 cells in nude mice abdominal cavity,which was reflected by the decrease of tumor number and tumor size.With the inhibition of miR-106a,the expression of TIMP2 in miR-antagomir group was significantly higher than that in miR-NC group (P＜0.05).Conclusion BGC-823 cell has the tumorigenicity in nude mice.Silencing of miR-106a inhibits gastric cancer cell metastasis, which suggests that it has the oncogenic function.MiR-106a may induce the strengthened peritoneal metastasis ability of BGC-823 cell through acting on TIMP2.

Objective:To discuss the effects of Buzhong Yiqi Decoction medicated serum combined with cisplatin on the expression of Nrf2 and its effects of downstream antioxidant proteins in human pulmonary adenocarcinoma A549/DDP cells.Methods:Totally 60 rats were divided into blank group, low-dosage group, medium-dosage group, and high-dosage group using a random number table method, with 15 rats in each group. The low-, medium-, and high-dosage groups were orally administered with 3.29, 6.57, and 13.13 g/kg Buzhong Yiqi Decoction for gavage, once a day, for 7 days. 1 hour after the last administration, blood was collected from the abdominal aorta to prepare the serum containing Buzhong Yiqi Decoction. A549/DDP cells were divided into 5 groups: blank group (rat serum), model group (rat serum+cisplatin), low-dosage group (low-dosage Buzhong Yiqi Decoction medicated serum+cisplatin), medium-dosage group (medium-dosage Buzhong Yiqi Decoction medicated serum +cisplatin), and high-dosage group (high-dosage Buzhong Yiqi Decoction medicated serum+cisplatin). And 24 h after the Buzhong Yiqi Decoction intervention, CCK-8 method was used to detect the IC 50 of cisplatin in each group. The fluorescence expression intensity of p-Nrf2 in A549/DDP cells of each group was determined by confocal fluorescence localization method. The mRNA relative expression of Nrf2 in A549/DDP cells of each group was detected by RT-qPCR. The relative expressions of Nrf2, p-Nrf2, GPX4, PRX1 and SOD in A549/DDP cells of each group were detected by Western blot. Results:Compared with the model group, the IC 50 of A549/DDP cells to cisplatin decreased in the low-, medium-, and high-dosage groups ( P<0.01); the fluorescence intensity of p-Nrf2 decreased ( P<0.01); the level of Nrf2 mRNA decreased ( P<0.01); the protein expressions of Nrf2, p-Nrf2, SOD, PRX1, and GPX4 were down-regulated ( P<0.01), with the effect being more significant in the medium- and high-dosage groups ( P<0.05). Conclusion:Buzhong Yiqi Decoction medicated serum can improve cisplatin resistance in human pulmonary adenocarcinoma cancer by inhibiting the active expression of Nrf2 in A549/DDP cells and down-regulating the target genes of SOD, PRX1 and GPX4 in vivo.

Objective To investigate the effects of high-risk human papillomavirus 16 E6 protein(HPV16 E6 protein)on invasion and migration of cervical cancer SiHa cells via regulating the expression of expression miR-23a.Methods Tissue samples from 100 patients with cervical cancer HPV-negative,100 HPV-positive patients,and 100 paracancerous normal tissues were collected;cervical cancer SiHa cells were divided into blank group,E6 overexpression group,negative transfection group,and E6 + miR-23a mimics group.The expression of miR-23a and HPV16 E6 mRNA were detected by qRT-PCR;MTT assay was used to detect the cell proliferation inhibition rate;flow cytometry to detect the apoptosis;Transwell chamber assay to detect cell invasion,and scratch test to detect the ability of cell migration.The expression of HPV16 E6,apoptosis related proteins(Caspase-3,Bax,Bcl-2),and migration related proteins(MMP-2,MMP-9)was detected by WB.Results The expression level of miR-23a was decreased in cervical cancer tissues,and that was lower in HPV positive cervical cancer tissues.Overexpression of E6 decreased the expression level of miR-23a,cell proliferation inhibition rate,apoptosis rate,Caspase-3 and Bax protein expression,and increased the expression of Bcl-2 protein,scratch healing rate,inva-sion cell number,MMP-2,MMP-9 protein expression(P<0.05);miR-23a mimics reversed the effects of E6 overexpression on the above indicators.Conclusion HPV16 E6 promotes the invasion and migration of cervical cancer cells,which may be related to the regulation of miR-23a expression.

·AIM:To investigate and compare the application of two screening models in the detection of retinopathy of prematurity (ROP). ·METHODS: The clinical data of 600 premature infants (1200 eyes) who underwent screening of eye diseases in the Department of Ophthalmology during the period from January 2016 to January 2017 were analyzed retrospectively. The fundus lesions were examined with binocular indirect ophthalmoscope (BIO) and the third generation of wide-angle digital retinal imaging system (RetCam Ⅲ). The examination results and adverse events during operation were statistically analyzed. ·RESULTS:In 1200 eyes of 600 patients,the probabilities of ROP detected by BIO and RetCam Ⅲ were 10.92% and 10.75%, respectively (P>0.05). With BIO as the golden standard, the accuracy, sensitivity, specificity, positive predictive value and negative predictive value of RetCam Ⅲ in examining ROP were 98.67%, 93.13%, 99.35%, 94.57% and 99.16%, respectively. There was no significant difference in the stage of ROP detected by BIO or RetCam Ⅲ (P>0.05). The probabilities of non-ROP lesions examined by BIO and RetCam Ⅲ were 4.83% and 4.58%, respectively (P>0.05). With BIO as the golden standard, the accuracy, sensitivity, specificity, positive predictive value and negative predictive value of RetCam Ⅲ in examining fundus non-ROP diseases were 99.67%, 94.83%, 99.91%, 98.21% and 99.74%, respectively. During the screening of BIO and RetCam Ⅲ,there were 17 cases (2.83%) and 7 cases(1.17%) with adverse events, respectively (P<0.05).·CONCLUSION: The examination results of RetCam Ⅲ are basically the same as those of BIO for ROP and non-ROP diseases. However,RetCam Ⅲ has more advantages in reducing adverse events during operation.

Objective To explore the current status of traditional Chinese medicine(TCM)research on alopecia areata(AA)and arogenetic alopecia(AGA),analyzing the clinical syndrome and treatment characteristics,and providing reference for TCM clinical diagnosis and treatment.Methods Search for research literature on traditional Chinese medicine treatment of hair loss in CNKI,Wanfang,and VIP separately.Removing duplicates through Notexpress and using CiteSpace and VOSviewer software for bibliometric analysis.Furthermore,focusing on the RCT research of traditional Chinese medicine internal treatment for AA and AGA,the data mining methods were used to analyze the pathogenesis,treatment methods,and prescriptions of traditional Chinese medicine.Results There are a total of 2954 literature on the treatment of hair loss by traditional Chinese medicine,with clinical research being the main research type.Among them,the main pathogenesis of AA is liver and kidney deficiency,qi and blood deficiency,etc.Commonly used herbs includes Polygonum multiflorum,Rehmannia glutinosa,Chuanxiong,Poria cocos,and Ligustrum lucidum,while the pathogenesis of AGA is mainly dampness heat accumulation,yin deficiency dampness heat,etc.,with commonly used herbs such as Platycladus orientalis,Alisma orientalis,Poria cocos,hawthorn,and Coix seed.Meanwhile,AA and AGA both pay more attention to the use of tonifying drugs in their treatment.Conclusion There is a growing trend in literature on the treatment of hair loss with TCM,and there are differences in the pathogenesis and treatment methods between AA and AGA in traditional Chinese medicine.It is worth further reference in the prescription and medication of clinical treatment of hair loss.

PURPOSE: The purpose of this study was to investigate the function of Zinc finger protein 488 (ZNF488) in nasopharyngeal carcinoma (NPC). MATERIALS AND METHODS: The endogenous expression of ZNF488 in NPC tissues, normal nasopharyngeal epithelium tissues and NPC cell lines were detected by quantitative reverse transcription polymerase chain reaction. ZNF488 over-expressing and knock-down NPC cell line models were established through retroviral vector pMSCV mediated over-expression and small interfering RNA (siRNA) mediated knock-down. The invasion and migration capacities were evaluated by wound healing and transwell invasion assays in ZNF488 over-expressing and control cell lines. Soft-agar colony formation and a xenograft experiment were performed to study tumorigenic ability in vitro and in vivo. Immunofluorescence and western blotting analysis were used to examine protein changes followed by ZNF488 over-expression. Microarray analysis was performed to explore gene expression profilings, while luciferase reporter assay to evaluate the transcriptive activity of Tcf/Lef. RESULTS: ZNF488 was over-expressed in NPC tissues compared with normal tissues, especially higher in 5-8F and S18, which are well-established high metastatic NPC clones. Functional studies indicate that over-expression of ZNF488 provokes invasion, whereas knock-down of ZNF488 alleviates invasive capability. Moreover, over-expression of ZNF488 promotes NPC tumor growth both in vitro and in vivo. Our data further show that over-expression of ZNF488 induces epithelial mesenchymal transition (EMT) by activating the WNT/beta-catenin signaling pathway. CONCLUSION: Our data strongly suggest that ZNF488 acts as an oncogene, promoting invasion and tumorigenesis by activating the Wnt/beta-catenin pathway to induce EMT in NPC.
Blotting, Western ; Carcinogenesis* ; Cell Line ; Clone Cells ; Epithelial-Mesenchymal Transition* ; Epithelium ; Fluorescent Antibody Technique ; Gene Expression Profiling ; Heterografts ; Luciferases ; Microarray Analysis ; Oncogenes ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Small Interfering ; Wnt Signaling Pathway* ; Wound Healing ; Zidovudine ; Zinc Fingers