Objective To explore the prognosis of elderly patients suffered from hospital-acquired pneumonia (HAP) by T cell subsets and clinical pulmonary infection score (CPIS).Methods A cohort of 125 elderly patients admitted in ICU & ED (Emergency Department) from Aug,2012 to Jul,2013 were enrolled for a prospective and observational study.The patients were divided into 3 groups:HAP survival group (n =50,group A),HAP death group (n =40,group B) and non-HAP group (n =35,control group).The criteria of exclusion were patients with auto-immune diseases,immunodeficiency,allergic disorders,malignancies,diabetes,trauma,surgical diseases,or patients with recent use of immunosuppressive agents or cyclooxygenase-inhibitors (Aspirin etc.).In the control group,patients with nosocomial pneumonia and other diseases afecting the CPIS were excluded.APACHE Ⅱ scores of all patients were recorded.Blood T cell subsets (including values of CD3,CD4 +,CD8 +,and CD4 +/CD8 +)were measured on the admission day,the 1st day of HAP onset and the 5th day after onset of HAP in HAP patients whereas these measurements were tested only on the admission day in controls.Meanwhile,the CPISs were recorded on the admission day,the 1st day of HAP onset and the 5th day after onset of HAP in HAP patients.Flow cytometer (FCM) was used to detect T cell subsets.Data of statistical analysis were represented as Mean ± SD.The significant differences in T cell subsets and CPIS between survival group and death group were analyzed by independent t test.The paired samples t test was employed in survival group and death group.Linear correlation analysis was made between CD4 +/CD8 + ratio and CPIS in survival and death groups,respectively.Results There were no significant differences in demographics and clinical features (including age,sex,length of stay,APACHE Ⅱ scores) of patients in survivors and non-survivors (P ＞ 0.05).The values of CDs (CD3,CD4 + and CD4 +/CD8 + ratio) between patients of control group and patients of HAP groups were not significantly different on the admission day (P ＞ 0.05).The values of CDs on the admission day were much lower than those on the 1 st day of HAP onset in both survivors and nonsurvivors (P ＜ 0.05).The values of CDs on the 5th day after onset of HAP were higher than those on the 1 st day of HAP onset in the survival group (P ＜ 0.05),while there were no significant differences in CDs between different intervals after HAP onset in the death group (P ＞ 0.05).There were no significant changes in values of CD8 + in any group (P ＞ 0.05).Both survivors and non-survivors had much higher CPIS values on the 1st day of HAP onset than those on the admission day (P ＜0.01).The survival group had higher CPIS on the 5th day after onset of HAP compared to the 1st day of HAP onset (P ＜0.01),while there was no significant change in the death group.Linear correlation analysis showed negative correlation between CD4 +/CD8 + ratio and CPIS on both the 1 st day of HAP onset (survival group:R =-0.740,P =0.004 ; death group:R =-0.613,P =0.035) and the 5th day after onset of HAP (survival group:R =-0.639,P =0.009; death group:R=-0.686,P=0.021).Conclusions The hospital-acquired pneumonia appears as an immune imbalance disorder.The difference in CDs is a promising objective tool,aiding in prediction of prognosis of HAP in the elderly,the lower the CDs,the higher severity.The CD4 + / CD8 + ratio showed a negative correlation with CPIS.Monitoring of T cell subsets and CPIS may provide clinical value for the treatment of hospital-acquired pneumonia in the elderly.

Thirty patients with malignant obstructive jaundice were treated with biliary stent implantation+brachytherapy+conformal radiotherapy (study group; n=15) or biliary stent implantation alone (control group; n=15). Total bilirubin (TBIL) levels significantly declined within 1 month in both groups. However, at 6 months, TBIL values began to increase in the control group and continuously declined in the study group. Maximum tumor diameter increased in the control group, while decreased in the study group (remission rate, 13/15 ). As for the study group, the survival rate at 0. 5, 1, and 2 years was 15/15,14/15, and 10/15, respectively, higher than the control group (15/15,5/15,and 1/15) . Combining biliary stent implantation with brachytherapy and conformal radiotherapy might be a safe and effective treatment of choice for patients with malignant obstructive jaundice.

Objective:To screen the new candidate molecules interacting with protein kinase Wee1B by yeast two hybrid system, and to analyze their interaction with Wee1B in the early stage of mouse fertilized eggs by bioinformatics.Methods:The plasmid pcDNA3.1/V5-His-TOPO-Wee1B wild type encoding mouse Wee1B gene was used as template to construct bait plasmid pGBKT7 Wee1B and the bait plasmid pGBKT7-Wee1B was transformed into yeast competent cells at SD/Trp (SDO),SD/Trp/X-α-Gal (SDO/X)and SD/Trp/X α Gal/AbA plates (SDO/X/A)plates to detect the toxicity and self-activation ability of yeast and its expression in yeast using Western blotting method.The yeast cells containing pGBKT7-Wee1B were fused with human ovary cDNA library, the yeast plasmid transformation of Escherichia coli positive clones were sequenced after identified by yeast transformation.BLAST analysis was carried out in GenBank,and its effect on the development of mouse fertilized eggs was deduced according to the gene annotation.Results:The double enzyme digestion analysis and sequencing analysis results showed that the pGBKT7-Wee1B bait plasmid was successfully constructed.The plasmid was transformed into the yeast,and there were no clones in the SDO/X/A plates.The pGBKT7-Wee1B and pGBKT7 empty vectors were transformed into the yeast,the bacteria were inoculated on the SDO plates,and the clones were uniformly grown on the two SDO plates.The positive clones were picked and expanded in culture,the protein was extracted and Western blotting showed that pGBKT7 Wee1B was expressed in the yeast.The bait plasmids were fused with human ovary cDNA library and the positive clones inserted into the fragment were identified by PCR. Nine proteins which interacted with Wee1B protein kinase were screened out by sequencing and blast analysis,and the proteins which could be closely related to the development of mouse oocytes and the development of fertilized eggs were analyzed by bioinformatics analysis.Conclusion:Using the yeast two hybrid system from human ovary cDNA library,nine interacting proteins with Wee1B protein kinase are screened and these screened proteins may regulate mouse oocyte maturation and early embryo development through interacting with Wee1B.

Objective To explore the effect of paracrine extracts derived from human adipose stem cells on white matter injury of neonatal rats and to compare the difference of therapeutive effect between the cerebellum medulla oblongata pool injection and the jugular vein injection.Method A total of 73 three-day-old SD rats were chosen to establish the model of white mater injury.After 24 hours,the 73 rats were randomized into the experimental group (n =46) and the control group (n =27).Then the experimental group was reclassified into ventricular group (n =23) and intravenous group (n =23).In the ventricular group,the paracrine extracts of human adipose stem cells was injected locally into the cerebellum medulla oblongata pool injection,while the extracts was injected into the jugular vein in the intravenous group.The control group was reclassified ventricular control group (n =15) and intravenous control group (n =12),and equivoluminal saline was injected the same way as the experimental group.Frozen sections of the brain tissue from 3 rats of each experimental group one day after injection were stained with fluorescein-conjugated streptavidin to study the distribution of the extracts.The brain tissue of 3 rats from each subgroup 3 days after injection were stained with hematoxylin eosin (HE) to observe the pathomorphological changes.While 7 days later,myelin basic protein (MBP) of white matter which was obtained from 7 rats of each group was detected by immunofluorescence staining.28 days after injection,the remaining rats were assessed by neurobehavior tests.For the rats that died during the experiment,the same number of the rats would be substituted in this study.Result The paracrine extracts were found to transfer to the brain lesion area,and the amount of the extracts was more in the ventricular group.The results of the HE staining showed that the white matter injury was more severe in the ventricular control group,and extensive area of infarction were found in this group.White matter injury was mild in the experimental group,and the structure of the corpus callosum was more complete in the ventricular group.MBP semi-quantitative scores of the ventricular group (0.7 ± 0.3) and intravenous group (1.7 ± 0.3) were lower than those of ventricular control group (3.4 ± 0.4)and intravenous control group(3.3 ±0.3).And the MBP scores of ventricular group was significantly lower than that of intravenous group (P ＜ 0.05).The scores of the neurobehavioral tests of the experimental group were significantly higher than those of the control group,while the scores of the ventricular group were significantly higher than those of the intravenous group (P ＜ 0.05).Conclusion The paracrine extracts derived from adipose stem cells could improve the prognosis of white matter injury,and cerebellum medulla oblongata pool injection showed better curative effect than the jugular vein injection.

BACKGROUND:Microencapsulated cels are commonly used as a tool to overcome immune rejection after subarachnoid transplantation. However, the effect of microencapsulation on the secretion of human pheochromocytoma cels is unclear.
OBJECTIVE:To observe the growth and secretion of primarily microencapsulated cultured human pheochromocytoma cels in artificial cerebrospinal fluid.
METHODS: The human pheochromocytoma tissues were digested successively to isolate human pheochromocytoma cels that were then cultured in artificial cerebrospinal fluid. Primary cels were covered with alginate-polylysine-alginate microcapsules, and then the cel morphology was observed with inverted phase contrast microscope. Levels of met-enkephalin and norepinephrine in cel culture medium were detected by enzyme-labeled immunosorbent assay (ELISA). We used cel counting kit-8 colorimetric assay to obtain the growth curve of human pheochromocytoma cels in artificial cerebrospinal fluid.
RESULTS AND CONCLUSION:Microcapsulated human pheochromocytoma cels were in suspension and the process outgrowth increased slowly. Compared with non-microcapsulated cels, the proliferation rate of microcapsulated cels increased significantly. ELISA results revealed a significant increase in the levels of met-enkephalin and norepinephrine secreted from the microencapsulated cels compared to the non-microcapsule group. There was a wide variation in contents of met-enkephalin and norepinephrine from different tumors. These findings indicate that microencapsulated human pheochromocytoma cels can survive wel and have good secretion function in artificial cerebrospinal fluid, and human pheochromocytoma cels from different tumor tissues have stable secretory function.

Objective To investigate the role of tissue-resident memory T lymphocytes in the pathogenesis of psoriasis.Methods Clinical information was collected from 32 patients with progressive plaque psoriasis.Tissue specimens were obtained from both lesional and nonlesional psoriatic skin of all the patients,as well as from faded lesions in 9 of these patients.Tissue specimens from the normal skin of 10 healthy individuals served as the controls.Immunohistochemical staining was performed to detect the two characteristic surface markers CD69 and CDI03 on tissue-resident memory T lymphocytes and to analyze the status of these T lymphocytes at different stages of psoriasis.The results of immunohistochemical staining were compared by t test.Results The mean number of CD69+CD103+ T lymphocytes per high-power field was significantly higher in lesional skin than in nonlesional skin of the 32 patients (11.34 ± 7.60 vs.2.72 ± 4.20,t =8.46,P ＜ 0.01),but similar between psoriatic lesions in the 9 patients before and after subsidence (14.33 ± 2.21 vs.12.00 ± 4.58,t =1.98,P =0.08).There was no significant difference in the mean number of CD69+CD103+ T lymphocytes between nonlesional psoriatic skin and normal control skin (2.72 ± 4.20 vs.1.70 ± 2.98,t =0.71,P ＞ 0.05).Conclusion Tissue-resident memory T lymphocytes may play a role in the formation and recurrence of psoriatic lesions in patients.

Objective To investigate the relationship of CD4+ and CD8+ T cells with melanocytes in skin of patients with psoriasis,and to study their clinical significance.Methods Tissue specimens were obtained from both lesional and nonlesional skin of 29 patients with progressive psoriasis and 5 patients with regressive psoriasis,as well as from normal skin of 6 healthy individuals.Immunohistochemical staining was performed to determine the quantity and distribution of CD4+ T and CD8+ T cells,as well as the quantity of melanocytes and proportion of cells containing pigment granules in the basal layer of these specimens.Statistical analysis was carried out with the software SPSS 18.0 by one-way analysis of variance (ANOVA),least significant difference (LSD) test and Pearson correlation analysis.Results In patients with psoriasis,the mean number of CD4+ T cells per high-power (× 200) field was significantly larger in lesional skin than in nonlesional skin (epidermis:5.29 ± 4.66 vs.0,P＜ 0.05;dermis:77.50 ± 43.66 vs.9.67 ± 7.73,P＜ 0.05),so was the mean number of CD8+ T cells per high-power (× 200) field (epidermis:7.83 ± 6.27 vs.0.71 ± 1.20,P＜ 0.05;dermis:46.08 ± 34.26 vs.5.54 ± 4.43,P ＜ 0.05).A significant increase was also observed in the number of CD4+ and CD8+ T cells in lesional skin of patients with psoriasis compared with the normal control skin (both P ＜ 0.05).The lesional skin of patients with psoriasis also showed significandy increased number of melanocytes (103.45 ± 16.96),but decreased proportion of pigment granule-containing cells (7.45％ ± 3.86％) in the basal layer compared with nonlesional skin (43.62 ± 14.20,P＜ 0.05;43.10％ ± 14.91％,P＜ 0.05) and normal control skin (43.33 ± 14.02,P＜ 0.05;54.17％ ± 29.40％,P ＜ 0.05).There were no significant differences in either the mean number of CD4+ T cells,CD8+ T cells and melanocytes or the proportion of pigment granule-containing cells between nonlesional psoriatic skin and normal control skin (all P ＞ 0.05).The mean number of melanocytes was significantly higher in regressive psoriatic lesions than in white patches arising in subsided psoriatic lesions (P ＜ 0.05) and normal control skin (P ＜ 0.05),but similar between white patches and normal control skin (P ＞ 0.05),while the proportion of pigment granule-containing cells was insignificantly lower in regressive psoriatic lesions than in white patches (P ＞ 0.05),and significantly lower in regressive psoriatic lesions and white patches than in normal control skin (both P ＜ 0.05).Neither the number of CD4+ T cells nor that of CD8 + T cells was correlated with the number of melanocytes or the proportion of pigment granule-containing cells in progressive psoriatic lesions (both P ＞ 0.05),while the number of both CD4 + T cells and CD8 + T cells was positively correlated with that of melanocytes (r =0.46 and 0.56,respectively,both P ＜ 0.05),but uncorrelated with the proportion of pigment granule-containing cells in nonlesional psoriatic skin (both P ＞ 0.05).Conclusions In progressive psoriatic lesions,there is a significant increase in the number of CD4+ and CD8 + T cells as well as melanocytes in the basal layer,but a significant decrease in the proportion of pigment granule-containing cells.After subsidence of psoriatic lesions,both the number of melanocytes and proportion of pigment granule-containing cells gradually reach the levels in normal skin of healthy individuals.

Objective To observed the impact of selenium supplementation therapy on the thyroid perioxidase antibody(TPO-Ab) levels and serum oxidative stress[malondialdehyde(MDA, glutathione peroxidase(GPx), and superoxide dismutase(SOD)] in patients with Hashimoto′s thyroiditis. Methods 79 patients with Hashimoto′s thyroiditis were randomly divided into trial group(n=44) and placebo group(n=35) .The double-blind treatment was for 24 weeks. The thyroid hormone levels, serum TPO-Ab levels, and oxidative stress indexes(MDA, GPx, and SOD) of both groups were detected before and after treatment. Results (1)There was no change of thyroid hormone levels either before or after treatments of both groups(P>0.05). (2)TPO-Ab of the trial group decreased significantly after the treatment(P<0.05). While the placebo group has little change. Group with TPO-Ab≤200 IU/ml and the course≤1 year manifested the most obvious declines by 29.98% and 26.63% respectively. (3)The oxidative stress level of trial group significantly decreased after 24 weeks. There was significantly positive correlation between the oxidative stress indexes and TPO-Ab. However the placebo group was with little change. Conclusion Selenium supplementation may reduce the level of TPO-Ab titers and oxidative stress in patients with Hashimoto′s thyroiditis, especially for those with lower antibody titers and short course.

OBJECTIVE: To explore the detection of autoantibodies in serum of patients with coal workers' pneumoconiosis( CWP).METHODS: Eight hundred and three cases of stage Ⅰ CWP patients were selected as study subjects by random sampling method.Anti-nuclear antibody and anti-neutrophil cytoplasmic antibody( ANCA) in serum were detected by indirect immunofluorescence assay; myeloperoxidased efficiency( MPO) antibody,anti-mitochondrial M2 antibody( AMA-M2) and anti-cyclic citrullinated peptide( CCP) antibody were detected by enzyme-linked immunosorbent assay;rheumatoid factor( RF) was detected by enhanced immunoturbidimetry of latexa.Group analysis was conducted according to age,lung function,length of dust exposure and the nature of dust exposure collection.RESULTS: In the serum of 803 CWP patients,the positive rate of anti-nuclear antibody,AMA-M2,RF,anti-CCP antibody,ANCA and MPO antibody were 9.7%,7.5%,7.3%,4.0%,2.6% and 0.8% respectively; the karyotype distribution of 78 cases of anti-nuclear antibody positive specimens was spotted( 43.6%), cytoplasmic( 20.5%), homogenous( 7.7%) and nucleolus( 5.1%),with a titer of 1:100.The positive rate of anti-nuclear antibody in the > 70.0 years group was higher than that of ≤60.0 and ≤70.0 years group( P < 0.017); the positive rate of anti-nuclear antibody in the abnormal lung function group was lower than that of the normal group( P < 0.01); the positive rate of anti-CCP antibody in the dust exposure length > 30.0 years group was higher than that of ≤30.0 years group( P < 0.017); the positive rate of anti-CCP antibody in silica-exposed group was lower than that in the coal-exposed group( P < 0.01).CONCLUSION: The positive rate of antinuclear antibody,AMA-M2,RF and anti-CCP antibody in CWP patients were high.The positive rate of anti-nuclear antibody is associated with age and lung function.The positive rate of anti-CCP antibody is related to the duration and nature of dust exposure.

OBJECTIVE: To explore the changes of serum levels and significance of surfactant protein( SP) A and SP-D in patients with stage Ⅰ coal workers' pneumoconiosis( CWP). METHODS: A random sampling method was used to select 88 cases of stage Ⅰ CWP patients as the CWP group,50 cases of healthy underground miners with similar dust exposure history as the dust exposure group and 38 cases of ground workers without dust exposure history as the control group. The serum levels of SP-A and SP-D in 3 groups were detected by enzyme-linked immunosorbent assays. RESULTS: The levels of serum SP-A and SP-D in the CWP group and the dust exposure group were higher than that of the control group( P <0. 05). The serum level of SP-D in the CWP group was higher than that of the dust exposure group( P < 0. 01). The serum level of SP-D in the smoking CWP subgroup was lower than that of the non-smoking CWP subgroup( P < 0. 05).CONCLUSION: The abnormal serum levels of SP-A and SP-D were related to the development of stage Ⅰ CWP. Smoking might affect the serum level of SP-D in stage Ⅰ CWP patients.