Objective To construct the eukaryotic expression vector pEGFP-N1/IL-37b and analyze the expression of IL-37 gene in THP-1 cells. Methods Total RNA was extracted from human peripheral blood mononuclear cells ( PB-MCs) and the coding region of IL-37b gene was amplified by RT-qPCR. Then, the gene was cloned into pEGFP-N1 eu-karyotic expression vector. After transfected the recombinant plasmid into THP-1 cells, the expression of IL-37 was detec-ted by RT-qPCR and Western blot. Results Double restriction enzyme digestion and gene sequencing showed that IL-37b gene was correctly inserted into the eukaryotic expression vector pEGFP-N1. RT-qPCR and Western blot showed that the IL-37 expression level was increased significantly (P<0. 01) after transfection in THP-1 cells. Conclusions We successful-ly constructed a novel anti-inflammatory cytokine IL-37 eukaryotic expression vector pEGFP-N1/IL-37b, which lays a foun-dation for further study on IL-37 functions and its association with related diseases.

Objective To investigate the expression of antinuclear antibodies(ANA) ,anti‐cyclic citrullinated peptide antibody (anti‐CCP) in elderly patients with rheumatoid arthritis(RA) ,and analyze its relation with depression .Methods ANA ,anti‐CCP were detected in 58 elderly RA patients ,50 non‐RA and 50 healthy people .Self‐rating Depression Scale(SDS) was used to assess the incidence of depression .Results The positive rate of anti‐CCP in RA group was significantly higher than that in control group and non‐RA group(P<0 .05) ,while there was no significantly difference in positive rate of ANA compared between RA group and non‐RA group(P>0 .05) .The positive rates of anti‐CCP and ANA in active stage were significantly higher than those in stable stage(P<0 .05) .The positive rates of anti‐CCP and ANA in RA complicated with depression were significantly higher than those in RA without depression(P<0 .05) .Multiple linear regression analysis showed that positive rates of anti‐CCP and ANA were all in‐dependent influence factors of depression in elderly RA patients(P<0 .05) .Conclusion Anti‐CCP and ANA in elderly RA patients over express ,and the positive rates are positively related with disease activity and depression .

Objective To describe the pathological unit and octagonal en bloc resection for the treatment of ossification ligamentum flavum(OLF)in thoracic spine with spondylotic myelopathy.Methods Ninety-five patients from January 2002 to January 2007 were diagnosed as thoracic OLF,61 males and 34 females with an average age of 53.9 years(range,31-78 years).There were upper thoracic spine OLF in 32 cases,middle thoracic spine OLF in 24 cases and lower thoracic spine OLF in 39 cases.Single-segment OLF was found in 53 cases,double segments OLF was found in 38 cases and three segments OLF was found in 4 cases.CT scan multiplanar co-localized reconstruction was employed to detect the structure of spine with OLF.The Japanese Orthopaedic Association(JOA)lower limb motor function score,sphincter function score and motor function improvement rate were used to evaluate the outcomes.Results CT scan was engaged to observe 141 OLF pathological unite.The OLF pathology unit was defined as all the spine structures between the extension lines of the lower margin of the OLF two adjacent pedicles.Each OLF associates with an OLF pathology unit.The mean follow up duration was 38.3 months(range,24-60 months).Among 86 patients with sensations disturbance before operation,67 totally recovered and 19 relieved after operation.Trunk restrictions in 69 cases before operation were completely recovered after operation.Postoperative JOA sphincter function score was 2.651±0.334,comparing with preoperation score(2.262±0.561),and the difference was statistically significant.Postoperative JOA motor function score was 3.694±0.429,which was significantly increased than preoperative score 1.539±0.873,and motor function recovery rate was 87.57%.There was excellent in 71 cases,good in 17 cases and fair in 5 cases.The excellent and good rate was 94.74%.Conclusion The octagonal en block resection is relative safe for treatment thoracic OLF with myelopathy.Pathological unit of OLF in thoracic spine is more accurate to summarize the pathological contents and features of the OLF and its adjacent structure.

Three butylphthalide derivatives were isolated from the Rhizome of Ligusticum chuanxiong using a series of isolation and purification approaches including macroporous resin, ODS-A column, Sephadex LH-20 and preparative HPLC. These structures were elucidated based on extensive spectroscopic data (UV, IR, HR-ESI-MS and NMR) and identified as (3Z,3aE)-(6R,7R,2'S)-6-hydroxy-7-(2-carboxyl-2-hydroxyethylthio)-3-(2-hydroxybutylidene)-4,5,6,7-tetrahydro-phthalide (1), (3Z,3aZ)-3-butylidene-6,7-dihydroxy-4,5,6,7-tetrahydro-phthalide 7-O-α-D-glucopyranosyl-(1→2)-β-D-fructo-furanoside (2) and 3-(3-β-D-glucopyranosyloxy-butylidene)-7-hydroxy-phthalide (3).

Objective To explore the changes in inflammatory reactions in tail-suspension mice infected by Klebsiella pneumoniae from spaceflight.Methods Tail suspension was used to simulate the physiological effects of microgravity.C57BL/6 mice were randomly divided into control (Con),control+K.pneumoniae T16-169 (Con+T16-169),tail suspension (TS) and tail suspension+K.pneumoniae T16-169 (TS+T16-169) groups.The level of inflammatory cytokines TNF-α,IL-6 and IL-1β mRNA in lung tissue and the plasma cytokine concentration were detected by RT-qPCR and xMAP technology,and HE staining was used to represent the morphological changes in lung tissue.Results Compared with the control group,the expression of inflammatory cytokines in lung tissue and plasma concentrations of all experimental groups were increased,and the difference in TS+T16-169 group was the most significant (P<0.01 or P<0.001).HE staining showed that the lung tissues in Con+T16-169 and TS+T16-169 groups were damaged in different degrees,and the damage of TS+T16-169 group was the most serious.Conclusion The K.pneumoniae from spaceflight significantly increases the expression of inflammatory cytokines in lung tissue and plasma concentrations after infecting tail-suspension mice,and induces more serious damages to the lung tissue,which suggests that inflammatory reactions can be increased in tail suspension mice infected by K.pneumoniae from spaceflight.

OBJECTIVETo investigate the effect of Chinese herbs for nourishing Shen-yin and removing Xiang-fire (NYRF) on estrogen receptor (ER) expression in uterus and ovary of rats contaminated with nonylphenol (NP) or its bisphenol A (BPA) mixture, for exploring the action mechanism of NYRF in antagonizing the estrogen-mimetic activity of environmental endocrine disruptors (EEDs).

METHODSEEDs contaminated female SD rats, 3-week old, were divided into two groups, the treated group fed with NYRF and the control group with corn oil during the same period of contaminating for 15 days. The wet weight (WW) and organ coefficient (OC) of uterus in rats, as well as the ER protein and mRNA expressions in rat's uterus and ovary were detected and compared.

RESULTSAs compared with normal range, WW and OC increased significantly in the contaminated rats of the control group, with significantly down-regulated ER protein expression in uterus, and expressions of ER alpha and ER beta gene and protein in ovary (P<0.05). While in the treated group, the above-mentioned abnormalities of various indicators were markedly reversed to a certain extent (P<0.05).

CONCLUSIONEEDs show estrogenic-mimetic action on productive organs, which could be antagonized by NYRF, resulting in the down-regulated mRNA and protein expressions of ER in reproductive organs, so as to reduce the sensibility of reproductive organs to EEDs, which is probably one of the acting mechanisms of NYRF.

Animals ; Drugs, Chinese Herbal ; pharmacology ; Endocrine Disruptors ; toxicity ; Female ; Ovary ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley ; Receptors, Estrogen ; metabolism ; Uterus ; drug effects ; metabolism

Generation 4 polyamidoamine (PAMAM) dendrimer was PEGylated with polyethylene glycol (PEG) at an average molecular weight 5 000 via amide bond. PAMAM and PEGylated PAMAM (PAMAM-PEG) dendrimer were used as drug nanocarriers. Methotrexate (MTX), an antineoplastic agent, was selected as a model drug. PAMAM/MTX and PAMAM-PEG/MTX complexes were prepared. The pharmacokinetic characters and anti-tumor activity of the PAMAM-PEG/MTX complex were studied as compared with MTX injection and PAMAM/MTX complex by intravenous injection in rats and S180 tumor bearing mice, separately. The plasma samples from normal rats were analyzed by HPLC method, and concentration-time data were analyzed using a non-compartmental analysis. Their anti-tumor effects in vivo were evaluated against S180 solid tumors in mice by measuring average tumor weight and calculating the inhibitory rate of tumor on day 17 after successive injections. The results showed that both plasma half-life and mean retention time (MRT) of the complexes were longer than that of MTX injection (P<0.01), while the area under the plasma concentration vs time curve (AUC) of PAMAM-PEG/MTX was the largest as compared with that of free drug and PAMAM/MTX complex (P<0.01). The inhibitory rate of tumor of PAMAM-PEG/MTX complex enhanced 2.1 and 1.8 times over that of free drug and PAMAM/MTX complex, respectively, indicating that PAMAM-PEG/MTX exhibited the highest antitumor activity. In summary, PEGylated PAMAM could be useful as a potential drug delivery carrier.
Animals ; Antimetabolites, Antineoplastic ; blood ; pharmacokinetics ; pharmacology ; Area Under Curve ; Cell Line, Tumor ; Dendrimers ; chemical synthesis ; pharmacokinetics ; Drug Carriers ; Female ; Male ; Methotrexate ; blood ; pharmacokinetics ; pharmacology ; Mice ; Neoplasm Transplantation ; Nylons ; chemical synthesis ; pharmacokinetics ; Polyethylene Glycols ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Sarcoma 180 ; pathology ; Tumor Burden ; drug effects

Objective To explore the effects of amniotic lacrimal duct stenting on the prevention of dry eye in castrated rabbits.Methods Thirtysix healthy male rabbits were selected,the third eyelid were cut off and antiinfection treatment were given,which were randomly divided into 3 groups (12 cases in each group),the castrated male rabbits models were made.Among them,group A was negative control group,group B was dry eye model group,group C was group of lacrimal amniotic membrane group.At 2 weeks before implantation of amniotic lacrimal duct stent,2 weeks,4 weeks and 6 weeks after implantation,the fluorescent (FL) examination,Western blot,Schirmer I examination,immunofluorescence staining and corneal confocal microscopy were performed.Results The levels of tear secretion and FL in the three groups among different time points were significantly different (F=7.126,P =0.009;F =9.658,P =0.016),and there were significant differences among three groups (F =12.582,P =0.005;F =13.187,P =0.013).The tendency of tear secretion and FL in the three groups were also significantly changed (F =8.531,P =0.007;F =10.652,P =0.019).The epithelial basal cells at 6 weeks after implantation in three groups were 3811 ±414,3820 ± 314,2789 ± 353,and the density of inflammatory cells was 266 ±28,266 ± 29,67 ± 13,there were significant differences among three groups (F =13.442,P =0.012;F =9.231,P =0.021).The K1 6 staining in the duct epithelium were negative,and the expression of α-SMA in the lacrimal duct tissue of group A,B and C was not changed at all time points after implantation of amniotic lacrimal stent,and there was no significant difference (F =14.681,P =0.002).Conclusion The amniotic lacrimal stent implantation has certain effect on the prevention of dry eye in rabbit.

Autophagy is implicated as an indispensable mechanism affecting multiple aspects of cell physiology,including sur-vival,differentiation and development. Emerging evidences also suggest autophagy is developing as a therapeutic strategy for various diseases,such as autoimmune diseases, inflammatory diseases, cancer, cardiovascular disease and neurological diseases. In this pa-per,we focus on the autophagy mechanisms in the occurrence and development of many diseases,and related interventions.This review provides new ideas and methods for prevention and treatment of diseases in clinics.

Objective To investigate the choroidal thickness (CT) in patient (＞ 50 years old) with central serous chorioretinopathy (CSC) compared to age-matched healthy controls.Methods Together 25 (＞ 50 years old) patients with CSC from January 2016 to January 2017 were enrolled in these study.Of 25 patients,20 patients were affected unilaterally and 5 patients were affected bilaterally.These patients were divided into affected eye group (30 eyes) and unaffected fellow eye group (19 eyes).Addition 26 age-matched healthy subjects were recruited as control group (26 eyes).Enhanced depth imaging technique of optical coherence tomography (EDI-OCT) was used to measure the choriodial thickness at 5 sites,including subfovea and 500 μm,1500 μm temporal and nasal to the fovea.Results The mean age of patients with CSC was (60.08 ± 7.68) years.The subfoveal choroid thickness of both affected eye group and unaffected fellow eye group was significantly thicker than that of the control group [(414.17 ±85.88)pan vs.(352.89 ±97.22) μn vs.(280.69 ±90.99) pau] (both P ＜0.05).In the patients with monocular affected eyes,the subfoveal choroid thickness of the affected eyes was thicker than that of the fellow eyes [(414.07 ± 85.88) μm vs.(352.89 ± 97.22) μm] (P ＜ 0.05).There were significant differences in choriodal thickness at other sites in the three groups (all P ＜ O.05).Conclusion The choriodal thickness in patients with CSC at over 50 years of age is significantly thicker than that in the agematched healthy subjects,with subfoveal choroid thickness ＞ 400 μm,which is helpful for the differential diagnosis of elderly patients with macular serous retinal detachment.