Biomimetics ; Blood Cells ; metabolism ; CD55 Antigens ; blood ; CD59 Antigens ; blood ; Cell Membrane ; Humans ; Male ; Submarine Medicine

AIM: To investigate the situation of patients with mechanical ocular trauma and the possible influencing factors related to the prognosis.METHODS: Totally 260 patients(286 eyes) with mechanical ocular trauma from January 2011 to January 2016 in Xiaogan Central Hospital were selected and retrospective studied.Sociology and clinical data including gender,age,career,injury causes,injury nature,treatment time and visual acuity on admission in these patients were univariate analyzed,then the positive factors were analyzed using Logistic regression.RESULTS: The middle-aged and young males were domain in 260 patients.The causes were mainly sharp instrument injury,while perforating injury of eyeball was the majority in injury nature.Univariate analysis showed that injury causes,injury nature,treatment time and eyesight on admission were influencing factors,while multivariate analysis by Logistic regression showed that visual acuity on admission and orbital contusion were independent influencing factors for patients ocular trauma.CONCLUSION: Ocular trauma is a majoy cause of monocular blind,there are close associations between prognosis of ocular trauma and orbital contusion and visual acuity on admission.But the majority of ocular trauma is preventable and treatable,the relevant measures should be developed to prevent the ocular trauma.

AlM: To analyze the correlation between tear film changes of allergic conjunctivitis and dry eye, then provide clinical references for better treatment.
METHODS: Fifty patients with allergic conjunctivitis were taken as the observation group, the control group was selected based on 1:1 case control theory, and we chose 50 health volunteers without ocular surface diseases, xerophthalmia and systematic diseases randomly, then fluorescein ( FL) staining, break-up time ( BUT) , Schirner l test ( SⅠt) , tear meniscus high ( TMH) and slit-lamp examinations were performed in the two groups.
RESULTS: ln the observation group, FL, BUT, Slt, TMH of right eyes and left eyes were statistically significant correlated ( P < 0. 05 ). FL and BUT were statistically significant between control group and observation group (P<0. 05). TMH and Slt has positive correlation, while FL and BUT has negative correlation and the differences were statistically significant (P<0. 05). CONCLUSlON: Due to inflammatory mediators participation, allergic conjunctivitis could lead to the stability changes of tear film which cause in dry eye. The stability changes of allergic conjunctivitis correlate to the damage degree of epithelium.

Objective: To observe the change of microglia activity after fast decompressing and/or hyperbaric oxygenation (HBO)-induced central nervous system (CNS) damage, so as to study the role of microglia in CNS dysbaric injury and the effects of HBO on microglia. Methods: Rats were randomly divided into the following groups: normal control, safe decompressing, fast decompressing (FD) injured, and HBO treated groups. Rat models of dysbaric injury were established by FD; 6 h later the rat models were subjected to HBO treatment. The activated microglia were detected by FITC-linked Isolectin B4; TNF-α and TNF-α converting enzyme (TACE) positive cells were detected immunohistochernically; and neural apoptosis was detected by TUNEL assay. TNF-α contents in CNS tissue were determined by ELISA and the bioactivity of sTNF-α in cerebrospinal fluid (CSF) were determined by L929 cell cytotoxicity bioassay. Results: 1134 positive microglia appeared in rats' CNS 6 h after FD treatment, peaked after 24 h, and declined thereafter. The activated microglia had morphological changes. Cell apoptosis indices of CNS reached its peak 48 h after FD treatment. Activated microglia and apoptotic neurons had similar distribution. TNF-α was detected in the brain and spinal cord 6 h after FD, significantly increased after 24 h, and peaked after 48 h. The content of TNF-α was positively correlated with IB4 positive cells and apoptosis index (P<0.05). TNF-α bioactivity in CSF of FD group had a similar change to TNF-α content in CNS tissue. The IHC results showed that, TNF-α and TACE positive cells had the same morphology and distribution to those of IB4 positive cells. HBO treatment significantly decreased IB4 positive cells after 24 h, 48 h, and 72 h; reduced TNF-α content in CNS tissues and TNF-α cytotoxicity in CSF; and decreased the apoptosis index after 48 h and 72 h. Conclusion: Microglial cells are quickly activated after dysbaric-induced injury of CNS. The activated microglia play a role in secondary injury through increasing TNF-α and TACE expression. HBO therapy can protect the neurons through depressing the activation and proliferation of microglia and reducing secretion of neurotoxin.

OBJECTIVE:To study the effects of Lactobacillus fermentum(Lb-f)on inhibition of cytoxan(CTX) on HCT116 cells of human colorectal cancer in vitro and in vivo. METHODS:HCT116 cells were cultured in vitro. MTT assay was used to in-vestigate the effects of 4,2,1μg/ml CTX and combined with 10μg/ml Lb-f on the survival rate of HCT116 cells. HCT116 cell xe-nograft tumor nude mice model was induced to investigate the effects of 100,50,25 mg/kg CTX and combined with 180 mg/kg Lb-f on tumor volume,tumor weight,relative tumor inhibition rate,the sensitization effect of chemotherapy (by q value),the number of peripheral blood leucocyte and platelet. RESULTS:Compared with CTX alone,CTX combined with Lb-f had no signifi-cant effect on survival rate of HCT116,relative inhibition rate of tumor volume in nude mice, relative inhibition rate of tumor weight and q value (P>0.05),but increased the number of peripheral blood leucocyte and platelet (P<0.05). CONCLUSIONS:No synergistic effects of Lb-f is found on the inhibition of CTX on the growth of HCT116 in vitro and in vivo;Lb-f can inhibit the decrease of peripheral blood leucocyte and platelet of HCT116 cell xenograft tumor nude mice induced by CTX.

Objective Detailed descriptions of the double-labeling immunofluorescence staining for fluorescence microscopy provides an ideal sample.Methods Rat liver frozen sections were used fixative were 95％ alcohol,95％ formaldehyde and acetone,frozen sections,with anti-CSE,Ki-67 polyclonal antibody and incubated with FITC,Cy3 fluorescence-labeled secondary fluorescently labeled secondary antibody staining,observed under a fluorescence microscope.Results Acetone fixed group visible in the proliferative phase (S phase) cells showed a red fluorescent nucleus,cytoplasmic green fluorescence.Conclusion The impact of double labeling immunofluorescence the effect of sample links and many factors,including the two most important factors are 2 and coordination with the primary antibody and select the appropriate fixative.

Objective To study the role of hydrogen sulfide (H2S) in the effect of SB203580 on proliferation and apoptosis of hepatic stellate cells and the effects of H2S on expressions of collagenⅠand collagenⅢmRNA in hepatic stel-late cells. Methods There were five groups of HSC-T6 cells in this study including control group (DMEM medium contain-ing10%fetal bovine serum), dimethyl sulfoxide (DMSO) group, sodium hydrosulfide (NaHS)group,SB203580 (SB)group and SB+NaHS group. MTT method was used to detect the cell proliferation and inhibition rate of HSC-T6 cells treated by SB203580 and H2S. The apoptotic rate of HSC-T6 cells was detected by flow cytometry with annexin V-FITC/PI double staining. RT-PCR was used to detect the expressions of collagenⅠand collagenⅢmRNA in HSC-T6. Results The apop-totic rate of HSC-T6 cells was significantly higher in SB group and SB+NaHS group than that of control group, and the rate was significantly higher in SB+NaHS group than that of SB group (P＜0.05). There was no significant difference in the apop-totic rate of HSC-T6 cells between DMSO and NaHS groups than that of control group. The expressions of collagenⅠand col-lagenⅢmRNA were found in five groups of cells. There was a higher expression of collagenⅠand collagenⅢmRNA in NaHS group than that of control group (P＜0.05). The expressions of collagenⅠand collagenⅢmRNA were significantly lower in SB group and SB+NaHS group than those of control group and NaHS group (P＜0.05). Conclusion H2S activated P38MAPK signal pathway. And P38MAPK was specifically blocked by SB203580 in HSC-T6 cells, which inhibited the cell proliferation stimulated by H2S and promoted the apoptosis.

To explore the role of the Chk2 protein expression and DNA double strand breaks (DSBs) repair in low dose hyper-radiosensitivity (HRS)/increased radioresistance (IRR) of non-small cell lung cancer, A549 cells were subjected to irradiation at the dosage ranging from 0.05-2 Gy. Clonogenic survival was measured by using fluorescence-activated cell sorting (FACS) plating technique. Percentage of cells in M-phase after low doses of X-irradiation was evaluated by phospho-histone H3-FITC/PI and Western blotting was used to detect protein expression of Chk2 and phospo-Chk2. DNA DSBs repair efficiency was also measured by induction and persistence of γ-H2AX. The results showed that the killing ability of irradiation with A549 cells increased at low conditioning dose below 0.3 Gy. Within the dose of 0.3 to 0.5 Gy, A549 cells showed a certain extent of radiation resistance. And when the dose was more than 0.5 Gy, survival fraction exhibited a negative correlation with the dosage. There was no difference between the 0.1 or 0.2 Gy dosage groups and the un-irradiated group in terms of the percentage of cells in M phase. But in the high dosage group (0.3-1.0 Gy), the percentage of cells in M phase was decreased markedly. In addition, the percentage of cells in M phase began to decrease two hours after irradiation. One hour after irradiation, there was no conspicuous activation of Chk2 kinase in 0.1 or 0.2 Gy group, but when the irradiation dose reached 0.3 Gy or higher, Chk2 kinase started to be activated and the activation level showed no significant difference among high dosage groups (0.4, 0.5, 1.0 Gy). Within 1 to 6 h, the DNA DSBs repair efficiency was decreased at 0.2 Gy but increased at 0.5 Gy and 1.0 Gy, which was in line with Chk2 activation. We are led to conclude that the mechanism of HRS/IRR in A549 cell line was probably due to early G(2)/M checkpoint arrest and enhanced DNA DSBs repair. In this regard, Chk2 activation plays a key role in G(2)/M checkpoint activation.

Objective To investigate the operation key points, instrument improvement and shortterm effects in total en bloc spondylectomy (TES) via a single posterior approach for thoracic and lumbar tumors. Methods A series of modified instruments have been designed for the TES, including threadwire saw (T-saw) with a diameter of 0.81 mm, director and clamping for the saw, L shape and furcation osteotomes.The corpectomy of original TES which was defined as "one step dissection" from anteriorly to posteriorly, was modified into "two step dissection" which means that corpectomy was performed with saw cutting anteriorposteriorly and the L shape cutting posterior-anteriorly. In the cases with difficulty in pediculotomy using a T-saw, furcation osteotome was used for pediculotomy. Ten patients with thoracic or lumbar tumors were treated with the modified TES. There were 1 case of bone giant cell tumor, 1 case of bone neurilemmoma and 8 cases of metastatic tumors. All patients suffered moderate-severe pain and neurological deficit. Results The average follow-up period was 8.1(3.3-18.1) months. The average operating time was 7.8 h(6.0-10.3 h),and average blood loss was 2100 ml (1200-3500 ml). No disruption of dural mater, the leakage of cerebrospinal fluid, iatrogenic spinal cord injury and major vessel damage occurred. Two patients who underwent pleura disruption happened during the operation were treated with intrathoracic drain remedy. Among 7 cases with thoracic tumors, significant improvement in neurological function were achieved in 5 patients with the improvement of one grade in ASIA classification, while no change was found in 2 cases. In 3 cases with lumbar tumor, lumbar nerve root pain relieved and the muscle strength had recovered to grade 4 at least postoperatively. Conclusion Significant improvement has been achieved in the maneuverability and safety of the modified surgical techniques in TES with a single posterior approach for thoracic and lumbar tumors.

The purpose of the study was to observe the effect of rapamycin (RAPA) on the differentiation and maturation of rat bone marrow-derived dendritic cells (BMDCs) in vitro. BMDCs from Wistar rats were cultured with granulocyte-macrophage colony-stimulating factor plus interleukin-4 in the presence or absence of RAPA (20 ng/mL), and stimulated with lipopolysaccharide (LPS) for 24 h before cells and supernatants were collected. Surface phenotype of BMDCs was flow-cytometrically detected to determine the expression of maturation markers, MHC class II and CD86. Supernatants were analyzed for the production of IL-12 and IFN-gamma cytokines by using ELISA. BMDCs were co-cultured with T cells from Lewis rats and mixed lymphocyte reaction was assessed by MTT method. The morphology of BMDCs stimulated with LPS remained immature after RAPA pretreatment. RAPA significantly decreased the CD86 expression, impaired the IL-12 and IFN-gamma production of BMDCs stimulated with LPS, and inhibited the proliferation of allogeneic T cells. In conclusion, RAPA can inhibit the maturation of BMDCs stimulated with LPS in terms of the morphology, surface phenotype, cytokine production, and ability of BMDCs to stimulate the proliferation of allogeneic T cells in vitro.