China ; Ethnic Groups ; genetics ; Female ; Humans ; Male ; Middle Aged ; Polymorphism, Genetic ; Receptors, Complement ; genetics

Background In vitro study showed that chemotaxis consist of chemotaxis factor 4(CXCR4)and stromal cells derived factor-1(SDF-1)and may play a role in the orientation and migration of retinal progenitor cells (RPCs)toward lesion.Overexpression of CXCR4 in RPCs can enhance the chemotaxis activity.Objective This work was to explore the feasibility and underlying mechanism of up-regulation of CXCR4 on RPCs induced by hypoxia.Methods RPCs were retained in an incubator with normal O2volume(16％)or hypoxia condition(10％ O2)for 12 hours and 24 hours respectively.Flow cytometer cell analysis screening(FACS)was conduced to measure the proportion of CXCR4-expressing cells,and CXCR4,HIF-1 mRNA were analyzed by reverse transcription-polymerse chain reaction(RT-PCR).The chemotical effect of 30 mg/L SDF-1 to RPCs cultured under the hypoxia condition was assessed using Boyden chamber.Results The expression level of CXCR4(CXCR4 mRNA/β-actin mRNA)inRPCs cultured by 10％ O2 for 12 and 24 hours were 0.28+0.07and 0.48+0.17 and increased by 1.75 and 3.00 fold more than that of 16％ O2 culture group(0.16+0.02)(P＜0.01).The expression level of HIF-1 mRNA(HIF-1 mRNA/β-actin mRNA)in RPCs cultured by 10％ O2 for 12 and 24 hours were 0.18 ±0.07and 0.38 ±0.13 and increased by 3.00 and 6.30 fold more than that of 16％ O2 culture group(0.06±0.01)(P＜0.01).The chemotical effect of 30 μg/L SDF-1 to RPCs increased from 13.00％ in 16％ O2 culture group to 36.00％ and 46.00％ in the cells cultured by 10％ O2for 12 and 24 hours.FACS revealed that the proportion of CXCR4+ cells in hypoxia-exposure for 12 and 24 hours were 26.90％ and 46.10％,respectively,but that in 16％ O2 culture group was 9.10％,showing a statistically significant difference(P ＜ 0.01).Conclusions RPCs induced by hypoxia can enhance the expression of CXCR4 in RPE cells and the chemotaxia to SDF-1.The overexpression of H1F-1 in RPCs may be involved in the up-regulation of CXCR4 expression.

Objective To investigate pulmonary function in terms of tidal respiration and the influencing factors on it in ＜ 34 weeks premature infants with RDS at corrected gestational age of 40 weeks.Methods A total of 49 of ＜ 34 weeks premature infants with RDS (RDS group) and 36 of ＜ 34 weeks premature infants without RDS (non-RDS group) followed throughout entire twelve months were collected fromn December 2013 to October 2015.Of them,35 RDS patients and 20 non-RDS patients had the pulmonary function examination.A total of 26 full term infants with hyperbilirubinemia (full term group) were recruited for comparison study.The differences in parameters of tidal respiration were compared among the three groups.The RDS patients were further divided into the mild RDS subgroup and severe RDS subgroup according to the severity of illness.Result ①The TPEF [(0.17 ± 0.04) s vs.(0.23 ± O.09) s],VPEF [(6.74±2.70) mLvs.(9.33±2.92) mL],TPEF/TE [(29.06±4.21)％ vs.(38.27± 7.16) ％],VPEF/VE [(32.54 ± 4.43) ％ vs.(39.64 ± 5.88) ％] in RDS group were significantly lower than those in full term group (P ＜0.05).The TPEF [(0.19 ±0.06) s vs.(0.23 ±0.09) s],TPEF/TE [(30.31 ±11.53)％ vs.(38.27±7.16)％],VPEF/VE [(34.39±8.44)％ vs.(39.64±5.88)％] in non-RDS group were significantly lower than those in full termn group (P ＜ 0.05).The TPEF,TPEF/TE,VPEF/VE in RDS group were lower than those in non-RDS group,but the differences were not significant (P ＞ 0.05).②The TPEF,VPEF,TPEF/TE,VPEF/VE in mild RDS group were higher than those in severe RDS group,but the differences were.not significant (P ＞ 0.05).③ Logistic regression analysis indicated that the gestational age was the protective factor of pulmonary function in premature infants with RDS.Conclusions Small airway resistance in ＜ 34 weeks premature infants with RDS is higher than that in full term infants.There was no significant difference in pulmonary function between RDS premature infants and non-RDS premature infants.The gestational age was the influencing factor of pulmonary function in premature infants with RDS.

Purpose To evaluate the clinical and pathological features of extramammary Paget’ s disease ( EMPD) and to investigate the related prognostic factors. Methods The clinical and pathological data of 30 patients with EMPD were reviewed retrospectively. The HE slides in the file were rechecked. Immunohistochemical stains were performed on archival paraffin-embedded sections with En-Vision method in 20 cases. Follow-up data were recorded. The relationship between the clinicopathological parameters and prognosis was statistically analyzed. Results Among the 30 patients, there were 26 males and 4 females, aged from 53 to 88 years with median age 70. Macroscopically, typical presentations of EMPD were eczematoid lesions. Microscopically, Paget’s cells were distributed sin-gly or in groups ( as glandular or nests patterns) within the epidermis. Immunohistochemical stains showed that Paget’ s cells were pos-itive for CK7, and negative for CK5/6 and p63. Follow-up data were available in 23 cases, 8 cases recurred resection, among them, 6 patients had positive resected margin and 4 died. The recurrent rate and mortality between the groups of dermal invasion and no der-mal invasion (P<0. 01 and P<0. 05), the groups of lymphatic metastasis and no lymphatic metastasis (P<0. 01), and the groups of positive and negative resected margin (P<0. 01 and P<0. 05) were statistically different. There were no statistically different with age and extent of lesion (P>0. 05). Conclusions CK7 is a sensitive and specific marker of Paget’s cells. Dermal invasion, lymphatic metastasis and positive resected margin are poor prognostic factors in EMPD.

In this paper, a colorimetric method with high sensitivity, reproducibility and low cost for determining glutamate decarboxylase (GAD) activity was developed based on Berthelot reaction. The optimum substrate system is 0.2 mol/L MacIlvaine buffer pH4.7, containing 0.1 mmol/L pyridoxal phosphate (PLP) and 10 mmol/L L-monosodium glutamate (L-MSG). The reaction mixture consisted of 200 ?L substrate solution and 1～100 ?L enzyme preparation from lactic acid bacteria and was incubated at 30℃. The enzymatic reaction was terminated by immersion in ice-water and addition of 200 ?L 0.2 mol/L sodium borate buffer, pH9.0, then 1 mL 6% phenol solution and 400 ?L sodium hypochloride were added. Color development was carried out in boiling water for 10 min then immediately put in ice-water bath for 20 min. The optical density is read at 630 nm. ?-aminobutyric acid (GABA) produced was calculated using the standard curve. Applications of the method in GAD studies were discussed.

A lipopeptide compound was isolated from the culture of Bacillus subtilis HSO121 by methods of acidic precipitation, solvent extract, fractional precipitation, adsorption and prepared thin-layer chromatography; and its molecular structure was determined by by ninhydrin assay and IR methods following the Amino analysis, MS-MS and ESI-MS. It shows that the isolated lipopeptide consists of two homologues with molecular mass 1,022D and 1,036D and bearing a cyclic structure with the amino acid sequence Leu-Leu-Asp-Val-Leu-Leu-Glu in the peptide chain, which indicates that the isolated lipopeptide falls into the analogs of surfactin.

Objective To explore how the severity of periventricular-intraventricular hemorrhage (PIVH)impact on physique and neurodevelopment in premature infants.Methods A total of 80 preterm infants with PIVH admitted to NICU of Qingdao Women and Children's Hospital from December 2013 to June 2015 were eligible.According to the Papile classification,the premature infants were divided into 4 groups.They were grade Ⅰ PIVH group,grade Ⅱ PIVH group,grade Ⅲ PIVH group and grade Ⅳ PIVH group.The infants with grade Ⅰ PIVH and grade Ⅱ PIVH belong to the low-grade PIVH group.The infants with grade Ⅲ PIVH and grade Ⅳ belong to the severe-grade PIVH group.All of them were regularly followed up for 12 months.Neurodevelopmental outcomes of infants at 6 and 12-month correction age were assessed by using the 20 items neuromotor assessment applying for 0-1 year old and the Bayley scales of infant development-Ⅱ.The differences in physical and neurophysical development of premature infants among 4 groups were compared.Results There were no significant differences in physical growth indicators such as body weight,body length and the incidence of weight growth retardation among 4 groups (all P＞0.05).The incidence of neurobehavioral abnormalities in infants with grade Ⅲ-Ⅳ PIVH was significantly higher than that of infants with grade Ⅰ-Ⅱ PIVH at 12-month correction age (21.05％ vs 3.28％,x2 =4.284,P=0.038).Physical development index(PDI) of grade Ⅰ-Ⅱ PIVH infants was significantly higher than that of grade Ⅲ-Ⅳ PIVH infants at 6-month correction age(F=11.500,P＜0.05).At 12-month correction age,grade Ⅰ-Ⅱ PIVH infants showed a significant higher mental development index(MDI) scores and PDI scores than those of grade Ⅲ-Ⅳ PIVH infants(F=14.227,16.515,all P＜0.05).Of the 80 cases assessed,infants with grade Ⅲ-Ⅳ PIVH had significantly higher rates of cerebral palsy(21.05％ vs 1.64％,x2 =6.300,P=0.012) and developmental delay (26.32％ vs 4.92％,x2=5.185,P=0.023) compared with grade Ⅰ-Ⅱ PIVH infants.Conclusions The severe PIVH can have negative effect on the neurodevelopmental outcomes of preterm infants and might induce mental retardation,cerebral palsy and other neurodevelopmental disabilities.Therefore,the regular follow-up and early intervention in preterm infants with PIVH should be implemented to improve the quality of their lives.

Adenosine receptors (AR) play an important role in the regulation processes for body temperature and vigilance states. During our previous studies, we noticed that aminophylline (a non-selective, blood-brain-barrier penetrably AR antagonist) could attenuate the effects of YZG-330 [(2R,3S,4R,5R)-2-(hydroxymethyl-5-(6-(((R)-1-phenylpropyl)amino)-9H-purin-9-yl)tetrahydrofuran-3, 4-diol] on lowering the body temperature. Hereby, we focused ourselves on the character of thermal regulation effect of YZG-330 in mice and tried to specify the receptor subtype via giving typical adenosine receptor antagonists. The results showed that both of the magnitude and lasting time of the effect that YZG-330 played on decreasing body temperature are in a dose-dependent manner: within the next 3 hour after intragastric administration (ig) of 0.25, 1 or 4 mg . kg-1 YZG-330, the extreme values on body temperature decreasing were (1.2 ± 0.3) °C, (3.6 ± 0.4) °C (P<0.001) and (7.4±0.5) °C (P<0.001), separately; whereas the duration that body temperature below 34 °C were 0, (10±5) and (153±4) min, separately. Adenosine A1 receptor (A1R) antagonist (DPCPX) could effectively reverse YZG-330's effect on decreasing body temperature, with intraperitoneal administration of DPCPX (5 mg . kg-1) 20 min prior than YZG-330 (4 mg.kg-1, ig), the extreme value on body temperature decreasing was (3.5 ± 0.7) °C (P<0.001), the duration that body temperature below 34 °C was (8±6) min (P<0.001). However, adenosine A2a receptor antagonist, SCH-58261, did not show any influence on the effects of YZG-330 at all. Combined with the fact that 8-SPT (a non-selective, blood-brain-barrier impenetrably AR antagonist) did not reverse the effect of YZG-330, we come to the conclusion that central-adenosine A, receptor plays a significant role on the thermal regulation effect of YZG-330.
Adenosine ; analogs & derivatives ; pharmacology ; Adenosine A1 Receptor Antagonists ; pharmacology ; Animals ; Body Temperature Regulation ; drug effects ; Mice ; Pyrimidines ; pharmacology ; Receptor, Adenosine A1 ; physiology ; Triazoles ; pharmacology ; Xanthines ; pharmacology

Adult ; Asian Continental Ancestry Group ; genetics ; Female ; Humans ; Male ; Middle Aged ; Peptidyl-Dipeptidase A ; genetics ; Polymorphism, Genetic

OBJECTIVETo study the effect of gene radiotherapy combining injection of recombinant plasmid pNEgr-mIL-12 with local X-irradiation on cancer growth and to elucidate the mechanisms of tumor inhibition.

METHODSAlkaline lysis was used to extract the plasmid and polyethylene glycol 8000 (PEG 8000) was applied for further purification of plasmids. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of IL-12 protein. C57BL/6J mice were subcutaneously inoculated with B16 melanoma cells and the plasmid was injected directly into the tumor. Gene-radiotherapy combining pNEgr-mIL-12 recombinant plasmid with X-irradiation was given three times to C57BL/6J mice bearing B16 melanoma. Changes in immunologic parameters of tumor-bearing mice were detected with relevant immunologic assays.

RESULTSResults showed a significant decrease in tumor growth rate (P<0.05-0.001) after 3 times of gene-radiotherapy with IL-12 and X-irradiation. Immunologic studies showed a significant increase in CTL and NK cytolytic activity (P<0.05-0.001) and an up-regulated secretion of IFN-gamma and TNF-alpha (P<0.01-0.001). Moreover, the expression of mIL-12 in B16 melanoma cells of the treated tumor-bearing mice was found to be higher than that of control.

CONCLUSIONpNEgr-mIL-12 plasmid combined with X-irradiation can increase tumor control and the mechanism of increased tumor inhibition is related to the enhancement of anticancer immunity in tumor-bearing mice.

Animals ; Combined Modality Therapy ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Therapy ; Interferon-gamma ; immunology ; Interleukin-12 ; biosynthesis ; genetics ; therapeutic use ; Killer Cells, Natural ; immunology ; Macrophages ; immunology ; Melanoma, Experimental ; immunology ; radiotherapy ; therapy ; Mice ; Mice, Inbred C57BL ; Plasmids ; Spleen ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Necrosis Factor-alpha ; immunology ; X-Rays