AIM: To investigate therapeutic efficiency of amniotic extraction on dry eye in rabbit model induced by topical benzalkonium chloride (BAC).
METHODS: Totally 26 rabbits (26 right eyes) with dry eye model were studied and divided into two groups:group A (control group with PBS eye drops, n = 13) and group B ( amniotic extraction group, n = 13). Another two rabbits were chosen as normal control. The SchirmerⅠ tests ( S Ⅰ t) and corneal fluorescein staining ( FL) were made, and the tear total protein content, amylase activity, lactoferrin, lysozyme contents, goblet cell density were performed in two groups before treatment and 1, 2, 4 and 8 wk after treatment.
RESULTS: There were significant differences in SIT, FL scores, lysozyme activity and goblet cell density among different groups at different time points (P<0. 05). But, there was no significant differences in SⅠt, FL scores, lysozyme activity and goblet cell density between two groups before treatment (P>0. 05). After 8wks' treatment with PBS, the mean differences of the group A showed great changes in SⅠt, lysozyme and goblet cell density compared with those before treatment ( P < 0. 05); but there was no significant differences in FL scores compared with those before treatment (P>0. 05). As for group B, 8wks after treatment, there were statistical changes in SⅠt, FL, lysozyme (P<0. 05); but there was no significant differences in goblet cell density compared with those before treatment ( P > 0. 05). It was evident that statistical differences were observed in S Ⅰ t, FL scores, lysozyme activity and goblet cell density between two groups at each time point (P<0. 05). However, there were no significant differences in total protein, lactoferrin, amylase activity at different time points (P>0. 05). Meanwhile there was no significant differences in total protein, lactoferrin, amylase activity between two groups before treatment ( P > 0. 05 ). But there were significant differences in total protein, lactoferrin, amylase activity between two groups after 4 and 8 wks'treatment (P<0. 05).
CONCLUSION: Amniotic extraction has significant therapeutic effect on the dry eye in rabbit model.

Argonaute 1 (AGO1) is a core component of the RNA-induced silencing complex (RISC) which plays a crucial role in small RNA-mediated gene silencing. AGO1 gene has been characterized in various plants, such as Arabidopsis and rice. However, there is no information about AGO1 in the medicinal plant species, Panax ginseng. Using the rapid amplification of cDNA ends technology (RACE), we cloned full-length PgAGO1 cDNA from Panax ginseng. It is 3 776 bp in length, including 204 bp of 5' UTR, 254 bp of 3' UTR, and 3 318 bp of ORF encoding 1106 amino acids. The molecular weight (MW) and theroretical isoelectric point (pI) of the deduced PgAGO1 protein is 122.22 kDa and 9.71, respectively. PgAGO1 shares 91.72% similarity with Arabidopsis AtAGO1 and contains three consered domains, including DUF1785, PAZ and Piwi, suggesting it is an authentic AGO. PgAGO1 was expressed in all of the tissues analyzed with the highest level in flowers and the lowest level in roots. The results provide useful information for further elucidating the function of AGO1 in Panax ginseng.
Amino Acid Sequence ; Argonaute Proteins ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Plant ; genetics ; Genes, Plant ; Molecular Sequence Data ; Panax ; genetics ; Plants, Medicinal ; genetics ; Sequence Alignment

OBJECTIVETo explore the mechanisms of electroacupuncture (EA) on postmenopausal osteoporosis (PMO).

METHODSSixty female SD rats aged 6 months were selected, resected double ovaries and fed for 90 days in order to make the model of experimental osteoporosis, and then, they were randomly divided into a model control group (without any therapy), a routine acupoints applying group, which were treated with EA at "Pishu" (BL 20), "Weishu" (BL 21), "Shenshu" (BL 23) and "Qihaishu" (BL 24), and an EA with tonification method (EAT) group at "Guanyuan" (CV 4) and "Zusanli" (ST 36), and an EA with dispersing blood stasis method (EAD) group at "Shenshu" (BL 23), "Geshu" (BL 17) and "Dazhu" (BL 11), and an EA with tonification and dispersing blood stasis method (EATD) group at "Guanyuan" (CV 4), "Zusanli" (ST 36), "Shenshu" (BL 23), "Geshu" (BL 17) and "Dazhu" (BL 11), 12 rats in each group. EA therapy was performed once a day, 30 min each time. Then these rats rested for 1 day after consecutive treatment for 6 days. The treatment lasted for 12 weeks. Enzyme linked immunosorbent assay (ELISA) was used to examine the level of TNF-alpha, and the level of E2, osteocalin (BGP) and carboxyterminal propeptide of type I procollagen (PICP), carboxyterminal telopeptide of type I lollagen (ICTP) by radioimmunoassay, and alkaline phosphatase by p-nitrophenylphosphate method, and Ca, P, Deoxypyridinoline (DPD) and creatinine (Cr) in urine were detected by automatic biochemistry analyzer.

RESULTSCompared with model control group, the level of DPD/Cr, Ca/Cr, and the serum contents of ALP, BGP, TNF-alpha and ICTP in all EA groups decreased significantly (all P < 0.01), the level of E2 and PICP in serum increased significantly (all P < 0.01). Compared with EATD group, the level of DPD/Cr and Ca/Cr in the other three EA groups had no statistical differences (all P > 0.05), but the level of ALP, BGP, TNF-alpha and ICTP in serum increased significantly (all P < 0.01), the level of E2 and PICP in serum decreased significantly (all P < 0.01).

CONCLUSIONAcupuncture can reduce significantly the level of DPD/Cr, Ca/Cr, and the serum contents of ALP, BGP, TNF-alpha and ICTP, increase significantly the level of E2 and PICP in PMO model rats and EA with tonification and dispersing blood stasis method is superior significantly to the other routine acupuncture methods.

Animals ; Bone and Bones ; metabolism ; Collagen ; metabolism ; Disease Models, Animal ; Electroacupuncture ; Female ; Humans ; Osteoporosis, Postmenopausal ; metabolism ; therapy ; Ovariectomy ; Rats ; Rats, Sprague-Dawley ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo explore the effects of survivin antisense RNA on taxol-induced apoptosis in leukemia cell line HL-60.

METHODSA survivin antisense eukaryotic vector pcDNA3-SVVas was transferred into HL-60 cells by electroporation. The live fraction was determined by trypan blue dye exclusion assay. Cell counting and MTT assay were performed to evaluate the sensibility of the transfected cells to taxol. Apoptosis was detected by DNA gel electrophoresis and nuclear staining.

RESULTSTwo positive cell clones, HL-60 SVVas and HL-60 neo were obtained. Compared to HL-60 and HL-60 neo cells, HL-60 SVVas cells growth was significantly reduced (P < 0.05). By MTT assay, the IC(50) of taxol to HL-60 SVVas, HL-60 neo and HL-60 cells were (14.4 +/- 1.87) ng/ml, (31.9 +/- 6.38) ng/ml and (32.0 +/- 3.52) ng/ml, respectively, the difference was significant by statistic analysis (P < 0.01). Agarose gel electrophoresis of genomic DNA from HL-60 SVVas showed typical DNA ladder, but DNA from HL-60 neo and HL-60 did not. Nuclei become condense in HL-60 SVVas cells.

CONCLUSIONSurvivin antisense RNA could enhance taxol-induced apoptosis in leukemia cell line HL-60. This may lay an experimental foundation for further research of gene therapy in leukemia.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Drug Synergism ; HL-60 Cells ; Humans ; Inhibitor of Apoptosis Proteins ; Microtubule-Associated Proteins ; genetics ; Neoplasm Proteins ; genetics ; Paclitaxel ; pharmacology ; RNA, Antisense ; genetics ; pharmacology ; Transfection

Objective To analyze the epidemiological characteristics of imported cases of dengue fever in Guangdong province (GD) and in I-long Kong (HK) during 2004-2006 to provide evidence for further cooperation in the prevention and control programs on dengue fever in the two places.Methods Descriptive statistical analysis was performed on data obtained from dengue fever surveillance and reporting network in GD and from Centre for Health Protection,Department of Health,HK.Results Both from GD and HK 44 and 93 imported cases of Dengue fever were reported during 2004-2006.Most patients from GD acquired their infection from Singapore (13 cases),Indonesia (9 cases) or Cambodia (6 cases) while patients in HK mainly were imported from Indonesia (31 cases),the Philippines (16 cases) and Thailand (15 cases).The peak seasons of the two places were both from July to September.During the non-peak season period,the number of cases in Hong Kong was higher than that in Guangdong.Male/ female ratio was 1.2 : 1 in GD and 1.1 : 1 in HK.Age of patients in GD appeared to range from 6-80 years,with 63.6% (28/44) of them aged 20-39 years.40.9% (18/44) of the cases were engaged in business,services,housework or unemployed.Those cases in HK were between 10-72 years of age,with 63.6%(28/44) of them aged 20-39 years while 47.3%(44/93) of the patients were with the occupation of business,services and industry.More cases in GD had a onset of disease before entering the border (27 : 17) than the cases in HK (35 : 57).The average time interval between onset and diagnosis were 7 and 9 days for GD and HK respectively.Conclusion Frequent travel between Southeast Asia in summer among the working class appeared to be the main factor,causing imported cases of dengue fever in GDand HK.It is crucial to provide health education targeted at these high risk groups in order to prevent importation of dengue fever in the two areas.

BACKGROUNDBerberine is one of the main constituents of Coptidis rhizoma (CR) and Cortex phellodendri. In this study, we investigated the beneficial effects of berberine on renal function and its possible mechanisms in rats with diabetic nephropathy (DN).

METHODSMale Wistar rats were divided into three groups: normal, diabetic model, and berberine treatment groups. Rats in the diabetic model and berberine treatment groups were induced to diabetes by intraperitonal injection with streptozotocin (STZ). Glomerular area, glomerular volume, fasting blood glucose (FBG), blood urea nitrogen (BUN), serum creatinine (Cr) and urine protein for 24 hours (UP24h) were measured using commercially available kits. Meanwhile, the activity of superoxide dismutase (SOD), content of malondialdehyde (MDA) in serum, activity of aldose reductase (AR) and the expression of AR mRNA and protein in kidney were detected by different methods.

RESULTSThe results showed that oral administration of berberine (200 mg x kg(-1) x d(-1)) significantly ameliorated the ratio of kidney weight to body weight. Glomerular area, glomerular volume, FBG, BUN, Cr and UP24h were significantly decreased in the berberine treatment group compared with the diabetic model group (P < 0.05). Berberine treatment significantly increased serum SOD activity and decreased the content of MDA compared with diabetic model group (P < 0.05). AR activity as well as the expression of AR mRNA and protein in the kidney was markedly decreased in the berberine treatment group compared with diabetic model group (P < 0.05).

CONCLUSIONThese results suggested that berberine could ameliorate renal dysfunction in DN rats through controlling blood glucose, reduction of oxidative stress and inhibition of the activation of the polyol pathway.

Aldehyde Reductase ; antagonists & inhibitors ; Animals ; Berberine ; pharmacology ; therapeutic use ; Diabetes Mellitus, Experimental ; complications ; Diabetic Nephropathies ; drug therapy ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Wistar ; Streptozocin

BackgroundChronic kidney disease (CKD) is closely related to the cardiovascular events in vascular calcification (VC). However, little has known about the characteristics of kidney injury caused by VC. Fibroblast growth factor 21 (FGF21) is an endocrine factor, which takes part in various metabolic actions with the potential to alleviate metabolic disorder diseases. Even FGF21 has been regarded as a biomarker in CKD, the role of FGF21 in CKD remains unclear. Therefore, in this study, we evaluate the FGF21 on the kidney injury in VC rats.

MethodsThe male Sprague-Dawley rats were divided into three groups: (1) control group, (2) Vitamin D3 plus nicotine (VDN)-induced VC group, (3) FGF21-treated VDN group. After 4 weeks, the rats were killed and the blood was collected for serum creatinine, urea nitrogen, calcium, and phosphate measurement. Moreover, the renal tissues were homogenized for alkaline phosphatases (ALPs) activity and calcium content. The levels of FGF21 protein were measured by radioimmunoassay. The levels of β-Klotho and FGF receptor 1 (FGFR1) protein were measured by enzyme-linked immunosorbent assay (ELISA). The structural damage and calcifications in aortas were stained by Alizarin-red S. Moreover, the structure of kidney was observed by hematoxylin and eosin staining.

ResultsThe renal function impairment caused by VDN modeling was ameliorated by FGF21 treatment, inhibited the elevated serum creatinine and urea level by 20.5% (34.750 ± 4.334 μmol/L vs. 27.630 ± 2.387 μmol/L) and 4.0% (7.038 ± 0.590 mmol/L vs. 6.763 ± 0.374 mmol/L; P < 0.01), respectively, together with the structural damages of glomerular atrophy and renal interstitial fibrosis. FGF21 treatment downregulated the ALP activity, calcium content in the kidney of VC rats by 42.1% (P < 0.01) and 11.7% (P < 0.05) as well as ameliorated the aortic injury and calcification as compared with VDN treatment alone group, indicating an ameliorative effect on VC. ELISA assays showed that the expression of β-Klotho, a component of FGF21 receptor system, was increased in VDN-treated VC rats by 37.4% (6.588 ± 0.957 pg/mg vs. 9.054 ± 0.963 pg/mg; P < 0.01), indicating an FGF21-resistant state. Moreover, FGF21 treatment downregulated the level of β-Klotho in renal tissue by 16.7% (9.054 ± 0.963 pg/mg vs. 7.544 ± 1.362 pg/mg; P < 0.05). However, the level of FGFR1, the receptor of FGF21, kept unchanged under VDN and VDN plus FGF21 administration (0.191 ± 0.0376 ng/mg vs. 0.189 ± 0.032 ng/mg vs. 0.181 ± 0.034 ng/mg; P > 0.05).

ConclusionsIn the present study, FGF21 was observed to ameliorate the kidney injury in VDN-induced VC rats. FGF21 might be a potential therapeutic factor in CKD by cutting off the vicious circle between VC and kidney injury.

OBJECTIVE: To investigate the clinical manifestations and laboratory features of B-ALL patients with EP300-ZNF384 fusion gene positive, so as to improve the understanding of this subtype disease. METHODS: The clinical data of 3 B-ALL patients with EP300-ZNF384 fusion gene positive admitted in Department of Hematology, the first medical center of Chinese PLA general hospital from February 2017 to February 2018 were collected and analyzed retrospectively. The clinical and laboratory characteristics as well as the therapentic outcome in B-ALL patients with EP300-ZNF384 fusion gene positive were analyzed. RESULTS: The fusion gene of EP300-ZNF384 was detected in 8.1%(3/37) of B-ALL patients. All cases showed the normal karyotype and aberrant CD13 and/or CD33 expression for immunophenotype. 3 patients were sensitive to traditional chemotherapy. CONCLUSION The B-ALL with EEP300-ZNF384 fusion gene positive may be a subgroup of B-ALL with a uniqe clinical characteristis and laboatorial features. EP300-ZNF384 positive patients show a good response to conventional chemotherapy, suggesting a favorable prognosis.

This study explored the mechanism of Sanhuang Decoction(SHD) in treating dextran sulfate sodium(DSS)-induced ulcerative colitis(UC) in mice with Candida albicans(Ca) colonization via high-throughput transcriptome sequencing. Specifically, the animal model was established by oral administration of 3.0% DSS for 7 days followed by intragastrical administration of Ca suspension at 1.0 × 10~8 cells for 4 days and then the mice were treated with SHD enema for 7 days. Afterwards, the general signs were observed and the disease activity index(DAI) was recorded every day. After mice were sacrificed, colon length and colon mucosa damage index(CMDI) were determined and the histomorphology was observed with the HE staining method. The fungal loads of feces were detected with the plate method. Anti-saccharomyces cerevisiae antibody(ASCA) and β-1,3-glucan in serum, and TNF-α, IL-1β, and IL-6 in serum and colon were detected by ELISA. High-throughput RNA sequencing method was adopted to identify transcriptome of colon tissues from the control, model and SHD(15.0 g·kg~(-1)) groups. Differentially expressed genes(DEGs) among groups were screened and the GO and KEGG pathway enrichment analysis of the DEGs was performed. The expression levels of NLRP3, ASC, caspase-1, and IL-1β genes related to the NOD-like receptor signaling pathway which involved 9 DEGs, were examined by qRT-PCR and Western blot. The results demonstrated that SHD improved the general signs, decreased DAI and Ca loads of feaces, alleviated colon edema, erosion, and shortening, and lowered the content of β-1,3-glucan in serum and TNF-α, IL-1β, and IL-6 in serum and colon tissues of mice. Transcriptome sequencing revealed 383 DEGs between SHD and model groups, which were mainly involved in the biological processes of immune system, response to bacterium, and innate immune response. They were mainly enriched in the NOD-like signaling pathway, cytokine-cytokine interaction pathway, and retinol metabolism pathway. Moreover, SHD down-regulated the mRNA and protein levels of NLRP3, caspase-1, and IL-1β. In a word, SHD ameliorates DSS-induced UC in mice colonized with Ca, which probably relates to its regulation of NOD-like receptor signaling pathway.
Animals ; Candida albicans/genetics* ; Colitis, Ulcerative/genetics* ; Colon ; Dextran Sulfate/toxicity* ; Disease Models, Animal ; Drugs, Chinese Herbal ; High-Throughput Nucleotide Sequencing ; Mice ; Transcriptome