Objective To investigate the changes of expression of Fas-associated proteins named Fas-associated death domain protein(FADD)and death-associated protein(Daxx)in the ischemic penumbra following transient focal cerebra ischemia in rats.Methods ①Adult male Sprague-Dawley rats were randomly divided into the sham-operated group and the cerebral ischemia model group.Rats underwent right middle cerebral artery occlusion(MCAO)for 2 h and reperfusion for 1,3,6,12 and 24 h using an intraluminal suture technique.The expression of FADD and Daxx mRNA and protein were measured with methods of immunohistochemistry.Western blot and reverse transcription-polymerase chain reaction(RT- PCR)respectively were used in the ischemic penumbra of rats.②Double-label fluorescence confocal laser scanning microscopy(CLSM)was performed to monitor FADD and Daxx intracellular location before and after ischemia.Results RT-PCR,Immunohistochemistry,Western blot experiments indicated that a very low level of FADD mRNA and protein were detected in the cerebral cortex of sham rats.The expression level both of FADD mRNA and protein increased significantly at 3 h after reperfusion,peaked at 12 h,then declined markedly at 24 h in the ischemic penumbra of model rats.RT-PCR,Immunohistochemistry indicated that a relatively high level of Daxx mRNA was detected in the cerebral cotex of sham rats.The expression level of Daxx mRNA increased significantly at 3 h after reperfusion and persisted to 24 h at a high level,whose protein had a same change of expression level in the ischemic penumbra of model rats. Immunofluorescence double-staining laser scanning by CLSM showed that the immunoreactivity of FADD was located in cytoplasm,and the intracellular translocation of the immunoreactivity of Daxx from nucleus to cytoplasm was monitored by measuring the green fluorescence after ischemia.Conclusion The transient upregulation of FADD and the persistant high level of expression of Daxx may contribute to neuronal apoptosis following cerebral ischemia/reperfusion.

Child ; Erythroblastosis, Fetal ; therapy ; Humans ; Infant, Newborn ; Male ; Rh Isoimmunization ; immunology

Objective To discuss the effects of multimodal combination dialysis on Klotho protein, fibroblast growth factor-23 (FGF-23) and brain natriuretic peptide (BNP) in patients with maintenance hemodialysis (MHD). Methods A randomized controlled trial (RCT) was conducted. 120 patients who was diagnosed with chronic renal failure (CRF) uremia receiving MHD over 3 months admitted to Blood Purification Centre of Department of Nephrology of the Second People's Hospital of Guiyang from December 2015 to December 2016 were enrolled, who were randomly divided into hemodialysis (HD) group (HD for 8 times a month), HD + hemofiltration (HF) group (HD for 8 times a month + HF once a month), and HD + HF + hemoperfusion (HP) group (HD for 8 times a month + HF for 4 times a month + HP once a month), with 40 patients in each group. Before and after treatment for 6 months and 12 months, blood was taken from venous circuit tube, the serum Klotho protein and FGF-23 levels were determined by enzyme linked immunosorbent assay (ELISA), and the serum BNP level was determined by electrochemiluminescence. Results 120 patients with MHD were enrolled in the final analysis without withdrawal. There were no significant differences in the levels of Klotho protein, FGF-23, or BNP before enrollment among the three groups (all P > 0.05). Compared with those before enrollment, the levels of serum Klotho protein after enrollment in three groups showed a sustained upward tendency, which were higher in HD + HF + HP group than in HD + HF group and HD group (μg/L: 2.59±0.61, 1.63±0.35, 1.13±0.26 at 6 months, F = 119.374, P = 0.000; 6.98±1.21, 3.57±1.03, 2.12±0.43 at 12 months, F = 275.675, P = 0.000); the levels of FGF-23 showed a sustained downward tendency, which were lower in HD + HF + HP group than in HD + HF group and HD group (ng/L: 69.22±38.26, 132.28±61.18, 178.50±74.64 at 6 months, F = 33.509, P = 0.000; 32.81±17.32, 87.93±43.27, 146.33±69.28 at 12 months, F = 55.466, P = 0.000);the BNP showed a similar tendency as FGF-23 (ng/L: 4083.39±2864.53, 7245.69±4643.81, 7969.12±5360.85 at 6 months, F = 8.758, P = 0.000; 1521.86±894.63, 4554.32±1969.84, 5013.89±2033.64 at 12 months, F = 49.003, P = 0.000). Conclusion Multimodal combination dialysis can increase the Klotho protein level, and decrease the levels of FGF-23 and BNP in MHD patients with CRF uremia.

AIMTo study the chemical constituents of the water-extracts of Selaginella tamariscina (Beauv.) Spring.

METHODSVarious chromatographic techniques were used to separate and purify the constituents. Their physico-chemical properties and spectral data were used to elucidate the structures.

RESULTSNine compounds were isolated and identified as (2R,3S)-dihydro-2- (3',5'-dimethoxy-4'-hydroxyphenyl)-7-methoxy-5-acetyl-benzofuran (1), 3-hydroxy-phenpropionic acid-(2'-methoxy-4'-carboxy-phenol) ester (tamariscina ester A, 2), sygringaresinol (3), 1-(4'-hydroxyl-3'-methoxyphenyl)glycerol (4), ferulic acid (5), caffeic acid (6), vanillic acid (7), syringic acid (8), and umbelliferone (9).

CONCLUSIONCompound 1 and 2 are new compounds, and the others were isolated from Selaginella for the first time.

Benzofurans ; chemistry ; isolation & purification ; Caffeic Acids ; chemistry ; isolation & purification ; Coumaric Acids ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Phenylpropionates ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Selaginellaceae ; chemistry ; Vanillic Acid ; chemistry ; isolation & purification

OBJECTIVETo investigate the interaction of deficiency in thrombosis-related gene in a mouse model.

METHODSTo generate mice carrying mutations in alpha-galactosidase A (Gla) and factor V Leiden (Fvl) and analyze the phenotypes, namely, tissue fibrin deposition and thrombus formation in organs.

RESULTSFibrin deposition in organs of mice carrying both mutations in Gla and Fvl was significantly increased compared with that in mice with single mutaton: [Gla(-/0) Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs.[Gla(-/0)Fv(+/+)]=(0.28+/-0.03)% vs.(0.07+/-0.007)%, P<0.01; [Gla(-/0)Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs.[Gla(+/0)Fv(Q/Q)+Gla(+/+)Fv(Q/Q)]=(0.28+/-0.03)% vs.(0.11+/-0.02)%, P< 0.01. Meanwhile, the number of thrombi on organ sections of mice carrying both mutations in Gla and Fvl was significantly increased compared with the single mutation carrier: [Gla(-/0)Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs.[Gla(-/0)Fv(+/+)]=1.9+/-0.7 vs. 0.0+/-0.0, P<0.05; [Gla(-/0)Fv(Q/Q)+Gla(-/-)Fv(Q/Q)] vs. [Gla(+/0)Fv(Q/Q)+Gla(+/+)Fv(Q/Q)]=1.9+/-0.7 vs. 0.3+/-0.1, P<0.05.

CONCLUSIONThese observations demonstrated that there was synergistic effect in Gla and Fvl deficiency in mice. It suggested that there could be a combination of GLA deficiency and FVL or other thrombosis-related gene defect in patients with genetic severe early-onset thrombosis.

Animals ; Factor V ; genetics ; Fibrin ; metabolism ; Immunohistochemistry ; Mice ; Mutation ; Thrombosis ; genetics ; metabolism ; alpha-Galactosidase ; genetics

OBJECTIVETo study the relationship between the infection of human papillomavirus (HPV) and external genital squamous cell carcinoma in situ.

METHODSHPV DNA was detected with the consensus primer by polymerase chain reaction (PCR) and its type was determined by restriction fragment length polymorphism (RFLP).

RESULTSHPV DNA was detected and HPV16 was the most frequently identified type in 29 (56.9%) of 51 cases of external genital squamous cell carcinoma in situ. HPV DNA was positive in 22 (55%) of 40 cases of bowenoid papulosis, among which 20 were of HPV16 type, one of HPV31 type, and one of HPV6 + 16 type. HPV DNA was positive in all 5 cases (100%) of Bowen's disease, among which 4 were HPV16 type and one of HPV6 + 16 type. HPV DNA was positive in 2 (33.3%) of 6 Queyrat erythroplasia cases and all were of HPV16 type.

CONCLUSIONHPV16 infection is strongly associated with the external genital squamous cell carcinoma in situ including bowenoid papulosis, Bowen's disease, and Queyrat erythroplasia.

Adult ; Aged ; Bowen's Disease ; virology ; Carcinoma in Situ ; virology ; Carcinoma, Squamous Cell ; virology ; DNA Probes, HPV ; DNA, Viral ; analysis ; Erythroplasia ; virology ; Female ; Genital Neoplasms, Female ; virology ; Genital Neoplasms, Male ; virology ; Humans ; Male ; Middle Aged ; Papillomaviridae ; isolation & purification ; Papillomavirus Infections ; virology ; Penile Neoplasms ; virology ; Polymorphism, Restriction Fragment Length ; Tumor Virus Infections ; virology ; Vulvar Neoplasms ; virology

OBJECTIVETo compare the effectiveness of immunofluorescence and polymerase chain reaction (PCR) in detecting human papilloma virus (HPV) in condyloma acuminata (CA).

METHODSHPVs in CA tissues from 60 patients were detected by immunofluorescence and PCR, respectively. Different subtypes of HPVs were also identified with restriction fragment length polymorphism (RFLP).

RESULTSThe positive detective rates of immunofluorescence and PCR were 56.67% (34/60) and 96.67% (58/ 60), respectively (P < 0.01). RFLP results showed HPV6 and HPV11 were the main subtypes in the detected virus, which accounted for 98.28%.

CONCLUSIONThe sensibility of PCR is superior to that of immunofluorescence.

Condylomata Acuminata ; virology ; Female ; Human papillomavirus 11 ; genetics ; isolation & purification ; Human papillomavirus 6 ; genetics ; isolation & purification ; Humans ; Male ; Microscopy, Fluorescence ; Polymerase Chain Reaction

AIMTo study the chemical constituents of the pine needles of Pinus massoniana lamb..

METHODSVarious chromatographic techniques were used to separate and purify. Their physico-chemical properties and spectral data (UV, IR, MS, 1H-1 H COSY, HMQC, DEPT, HMBC and ORD ect.) were measured for structure elucication.

RESULTSThree compounds were isolated from the n-BuOH fraction of water-extracts. Their structures were identified as massonianoside A (4), massonianoside A: (7S, 8R)-3, 4, 9'-trihydroxyl-3-methyoxyl-7, 8-dihydrobenzofunan-1'-propanolneoligan-9-O-alpha-L-rhamnopyranoside, massonianoside C (5), (7S, 8R)-9,9'-dihydroxyl-3,3'-dimethyoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-alpha-L-rhamnopyranoside and cedrusin-4-O-beta-glucoside (6), (7S, 8R)-3',9,9'-trihydroxyl-3-methoxyl-7,8-dihydrobenzofunan-1'- propanolneoligan-4-O-beta-D-glucopyranoside.

CONCLUSIONCompound 4 and 5 are new compounds.

Benzofurans ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Molecular Structure ; Pinus ; chemistry ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry

AIMTo study the chemical constituents from Corallodiscus flabellata.

METHODSThe compounds were isolated and purified by macroporous adsorption resin, silica gel column chromatography and identified on the basis of their physiochemical and spectral data.

RESULTSThree phenylethanoid glycosides (I-III) were obtained from the n-BuOH fraction of water-extracts. Their structures were elucidated as 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl (1-->3)]-beta-D-glucopyranoside (I), 3,4-dihydroxyphenylethanol-8-O-[4-O-trans-caffeoyl-beta-D-apiofuranosyl (1-->3)-beta-D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (II) and 3,4-dihydroxyphenylethanol-8-O-[beta-D-apiofuranosyl(1-->3)-beta- D-glucopyranosyl-(1-->6)]-beta-D-glucopyranoside (III).

CONCLUSIONCompounds I, II and III are new compounds.

Caffeic Acids ; chemistry ; isolation & purification ; Disaccharides ; chemistry ; isolation & purification ; Glycosides ; chemistry ; isolation & purification ; Magnoliopsida ; chemistry ; Molecular Structure ; Plants, Medicinal ; chemistry

AIMTo study the chemical constituents of Selaginella tamariscina (Beauv.) Spring.

METHODSVarious chromatographic techniques were used to separate and purify the chemical constituents. Their physico-chemical properties and spectral data were used to elucidate the structures.

RESULTSFour compounds were isolated from the n-BuOH fraction of the water-extracts. Their structures were identified as 1-hydroxy-2-[2-hydroxy-3-methoxy-5-(1-hydroxyethyl)-phenyl]-3-(4-hydroxy-3,5-dimethoxy)-propane-1-O-beta-D-glucopyranoside (tamariscinoside B, I), adenosine (II), guanosine (III), arbutin (IV).

CONCLUSIONTamariscinoside B (I) is a new compound, while the others were isolated from Selaginella for the first time.

Adenosine ; chemistry ; isolation & purification ; Arbutin ; chemistry ; isolation & purification ; Glucosides ; chemistry ; isolation & purification ; Guanosine ; chemistry ; isolation & purification ; Molecular Conformation ; Molecular Structure ; Plants, Medicinal ; chemistry ; Selaginellaceae ; chemistry