AIM: To study the infiltration of polymorphonuclear neutrophils ( PMNs ) after conjunctival flap covering in alkali-burned cornea.
●METHODS: Rabbit cornea alkali-burned model was made, then 50 rabbits were randomly divided into the experimental group ( n=25 ) and the control group ( n=25 ) . At the same time the surgery of conjunctival flap covering was given to rabbits of the experimental group. The condition developing of alkali-burned cornea was observed by slit lamp biomicroscopy, and took photos in two groups. The infiltration of PMNs was identified by hematoxylin eosin ( HE) staining in different periods.
●RESULTS:The quantity of PMNs increased on the 3d, reached the lower level on 7d, shown a peak on the 14d, then decreased gradually. PMNs level of the experimental group was significantly lower than that in the control group, and the difference of 3, 14 and 21d was significant (P<0. 05).
●CONCLUSION: During the wound healing process, alkali - burned cornea has close relation with the infiltration of PMNs. The treatment of conjunctival flap covering for the severe alkali-burned cornea was found to have good effect.

Objective To study the relationship of lung function and the diaphragmatic nerve electrophysiological changes in elderly patients with chronic obstructive pulmonary disease(COPD).Methods The phrenic nerve motor conduction(PNC),diaphragmatic motor evoked potentials(dMEP)and diaphragmatic compound muscle action potentials(dCMAP)were detected in 50 COPD patients and 50 healthy controls aged 60 years and over,respectively. Results (1)The lung function of elderly COPD patients were significantly less than that of control group(P0.05),the lower amplitude of dCMAP than the control(P

Adult ; Aneurysm, Dissecting ; complications ; Aortic Aneurysm ; complications ; Humans ; Lower Extremity ; physiopathology ; Male

Objective Endoscopic ultrasound guided fine needle aspiration biopsy (EUS-FNA) was performed for diagnosis and treatment of the pelvic Lesions and safety of this method were evaluated.Methods EUS-FNA was performed in 52 patients with pelvic Lesions selected by EUS,CT or MRI between March 2009 and June 2012.Both 19 and 22 gauge needles were used for EUS-FNA.The tissue specimens were analyzed by cytologic or histologic examination.The pelvic cystic lesions were drained and in which purulent lesions were lavaged with Metronidazole repeatedly.Results All patients recieved fine needle biopsy of the pelvic lesions.Among the 52 lesions,there were 42 solid lesions,10 cystic lesions.In solid lesions,cytology and pathology demonstrated malignant tumors in 28 patients,3 cases of malignant stromal tumors,3 cases of Inflammatory mass cases,2 cases of lymphoma,1 case of dermoid cyst,5 cases of other.In cystic lesions,2 cases of serous cystadenoma,perirectal abscess in 8 cases.6 purulent lesions were lavaged with Metronidazole repeatedly.Diagnosis rates of samples for immunohistology remained similar between 22 gauge and 19 gauge needles (P ＞0.05).During the operation,8 cases of perirectal abscess patients have different degree of pain.There were no other complications after the procedures except one patient suffering from fever.Conclusion EUS-guided FNA is minimaly invasive,a safe and accurate method for diagnosis of pelvic lesions.

Objective To detect the expression of humar telomerase reverse transcriptase (hTERT) and telomerase activity in HeLa,MCF-7,SMMC7721 PC-3m and U2OS cell lines. Methods The techniques of immunochemistry and TRAP-ELISA were employed to detect the expression of hTERT and telomerase activity in different tumor cell lines.Results The positive rate of hTERT in HeLa cells(93.75%?3.10%)was significantly higher than that in U2OS cells(2.75%?0.96%),besides the other three cell lines showed an positive rates of hTERT in MCF-7(92.50%?2.65%),SMMC7721 (53.75%?2.22%)and PC-3m(23.50%?2.89%); Meanwhile,the telomerase activity of HeLa cells (94.58%?3.49%) was also much higher than that of U2OS(3.02%?0.43%),likewise the telomerase activities of the other three cell lines (MCF-7,SMMC7721,PC-3m) were 73.90%?4.50%,66.67%?3.35% and 50.62%?1.96%, respectively. Conclusion The expression of hTERT and telomerase activity show obvious differences among five tumor cell lines,suggesting that telomerase inhibitors cannot effect on all the tumor cell lines.

Objective:To explore the effects of serum cystatin C (Cys C) and uric acid (UA) concentrations before treatment on the prognosis of small cell lung cancer (SCLC) patients.Methods:A total of 196 patients diagnosed with SCLC in Affiliated Hospital of Qingdao University from April 2015 to December 2018 were selected, and hematological indicators such as serum Cys C and UA before treatment were collected. The receiver operating characteristic (ROC) curve was used to determine the optimal cut-off values of Cys C and UA. The Kaplan-Meier method was used for survival analysis. Cox proportional hazards model was used for univariate and multivariate analysis.Results:The optimal cut-off values of serum Cys C and UA before treatment were 0.775 mg/L and 296.45 μmol/L, respectively. Survival analysis showed that with the optimal cut-off value, the median progression-free survival (PFS) of patients with high concentrations of serum Cys C and UA (5.49 months vs. 8.57 months, χ2=35.943, P<0.001; 6.67 months vs. 8.20 months, χ2=8.047, P=0.005) and overall survival (OS) (13.37 months vs. 23.95 months, χ2=21.355, P<0.001; 14.13 months vs. 20.97 months, χ2=11.333, P=0.001) were shorter than those of patients with low concentrations. Univariate analysis showed that factors related to PFS were smoking history ( HR=0.707, 95% CI: 0.518-0.965, P=0.029), staging ( HR=1.776, 95% CI: 1.329-2.373, P<0.001), first-line medication ( HR=1.596, 95% CI: 1.072-2.376, P=0.021), chest radiotherapy ( HR=2.407, 95% CI: 1.803-3.214, P<0.001), Cys C ( HR=3.602, 95% CI: 1.716-7.561, P=0.001), UA ( HR=1.002, 95% CI: 1.000-1.003, P=0.036), and alkaline phosphatase ( HR=1.010, 95% CI: 1.004-1.016, P=0.001); factors related to OS included smoking history ( HR=0.577, 95% CI: 0.382-0.870, P=0.009), staging ( HR=1.846, 95% CI: 1.295-2.630, P=0.001), chest radiotherapy ( HR=2.041, 95% CI: 1.426-2.921, P<0.001), Cys C ( HR=9.506, 95% CI: 3.278-27.564, P<0.001) and UA ( HR=1.003, 95% CI: 1.001-1.005, P=0.006). Multivariate analysis showed that chest radiotherapy ( HR=2.553, 95% CI: 1.774-3.672, P<0.001), Cys C ( HR=4.538, 95% CI: 1.875-10.982, P=0.001) and alkaline phosphatase ( HR=1.011, 95% CI: 1.005-1.018, P=0.001) were independent prognostic factors for PFS; Cys C ( HR=9.028, 95% CI: 2.680-30.413, P<0.001) was an independent prognostic factor for OS. Conclusion:Both serum Cys C and UA concentrations before treatment in SCLC patients have a certain relationship with the prognosis of the patients. Those with elevated concentrations have shorter PFS and OS and poor prognosis. The high concentration of serum Cys C before treatment may indicate a rapid progression of the disease and a short survival time. It is necessary to pay attention to disease progression and recurrence.

Objective To find methods to isolate and purify plasminogen activator (Pla) from artificial culture of Yersinia pestis. Methods Ultrasonication and urea extracting combined by ammonium sulfate salting-out were tried to extract Pla. High performance liquid chromatography(HPLC) was used to purify Pla. The first step was ion exchange and the second was gel filtration, Preparative electrophoresis was used to purify Pla, too. The enzyme activity of the isolated or purificated Pla was detected. Results Both 50% - 60% saturated ammonium sulfate deposition of supernatant of plague bacilli ultrasonication and 0 - 10% saturated ammonium sulfate deposition of supernatant of plague bacilli powder soaked by urea had three bands(Mr about 31×103, 35×103 and 37×103) and lysis rings were 6.5 and 7.2 mm in diameter respectively when the enzyme activity was detected. Pla purified by HPLC was mainly composed of three bands(Mr about 31×103, 35×103 and 37×103), occupying more than 80% of total protein weight and lysis ring was 5.0 mm in diameter. Pla purified by preparative electrophoresis mainly consisted of three bands(Mr about 31×103, 35×103 and 37×103) with other proteins of low concentration nearby, no lysis ring was detected. Conclusions Pla is collected by the methods of ultrasonication and urea extracting. Priliminary purification of Pla can be achieved by HPLC and preparative electrophoresis.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Female ; Fluorescence ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Mycobacterium tuberculosis ; isolation & purification ; Paraffin Embedding ; Polymerase Chain Reaction ; methods ; Retrospective Studies ; Tuberculosis, Pulmonary ; diagnosis ; microbiology ; Young Adult

OBJECTIVETo explore the effects of stromal interaction molecule 1 (STIM1) on the expression of apoptosis-related proteins in prostate cancer PC-3 cells.

METHODSWe transfected the lentivirus vector STIM1-pGCSIL-GFP carrying STIM shRNA into human hormone-independent prostate cancer PC-3 cells, and 3 days later observed the transfection efficiency by fluorescence microscopy. At 7 days after transfection, we determined the expression of STIM1 in the PC-3 cells by RT-PCR and Western blot and those of apoptosis-related proteins Bcl-2, Bax, survivin and activated Caspase-3 by Western blot.

RESULTSAt 3 days, inverted microscopy revealed a transfection efficiency of > 80%. At 7 days, the STIM1 expression was significantly inhibited at both mRNA and protein levels. The Bcl-2/Bax rate was remarkably decreased as compared with that of the control group (0. 31 vs 1.24 ) , and the survivin expression was markedly reduced, 0. 14 times that of the relative expression in the control. However, the Caspase-3 cleavage was significantly activated, 1.52 times that of the control (P <0.05).

CONCLUSIONSTIM1 can be regarded as an oncogene in prostate cancer PC-3 cells. Inhibition of its expression can induce PC-3 cell apoptosis by reducing the Bcl-2/Bax rate, decreasing the survivin expression, and activating the Caspase-3 pathway.

Apoptosis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Humans ; Inhibitor of Apoptosis Proteins ; metabolism ; Male ; Membrane Proteins ; genetics ; Neoplasm Proteins ; genetics ; Prostatic Neoplasms ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Small Interfering ; genetics ; Stromal Interaction Molecule 1 ; Transfection ; bcl-2-Associated X Protein ; metabolism

Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on cytokine expression in splenic dendritic cell (DCs). Method DCs isolated from the spleens of male Wistar rats were seed-ed on 96-well (1×10s cells/well) cell culture plates, and the cells were stimulated with HMGB1 for various length of time or in different concentrations. (1) The time-dependent response between HMGB1 and tumor necrosis factor-α(TNF-α) as well as interleukin-12 (IL-12) gene/protein expressions: 24 wells of DCs were dividedly into six groups including 24 h-normal controls (n=4), 48 h-normal controls (n=4), 72 h-normal controls (n=4), and 24 h-HMGB1 treated group (n=4), 48 h-HMGB1 treated group (n=4) as well as 72 h-HMGB1 treated group (n=4), respectively. Among three HMGB1-treated groups, DCs were stiraulated by 1 μg/mL HMGB1. DCs were denatured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvested to determine Il-12 as well as TNF-α protein levels. (2) The dose-dependent response between HMGB1 and TNF-α as well as IL-12 gene/protein expressions: 16 wells of DCs were dividedly into four groups in-cludingnormal controls (n=4), 0.1 μg/mL HMGB1 treated group (n=4), 1 μg/mL HMGB1 treated group (n =4), and 10 μg/mL HMGB1 treated group (n=4), respectively. After stimulated for 48 h, DCs were dena-tured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvest-ed to determine IL-12 as well as TNF-α protein levels. Total RNA was extracted from cells using the single-step technique of acid guanidinium thiocyanate-chloroform extraction according to the manufacturer' s instruction (Promega, Madison, WI). mRNA for TNF-α and IL-12 were quantified by SYBR Green two-step, real-time re-verse transeription-polymerase chain reaction taking glyceraldehyde-3-phesphate dehydrogenase (GAPDH) as an internal standard. Levels of IL-12 and TNF-α in cell culture supernatants were determined with ELISA, strictly fol-lowing the protocols provided by the manufacturer. Data were analyzed with a one-way ANOVA. A P-values <0.05 were considered statistically significant. Results After stimulation with 1 μg/mL HMGB1, IL-12 and TNF-α protein and gene expressions in rat splenic DCs were markedly up-regulated at 24 h to 72 h (P<0.05 or P<0.01), and the expression levels of IL-12 and TNF-α peaked at 48 h (P<0.01). When DCs were cultured in the presence of 0.1 μg/mL, 1 μg/mL, and 10 μg/ml HMGBI for 48 h, expressions of IL-12 and TNF-α were also significantly up-regulated (P<0.01), and values of these cytokines were highest in 1 μg/mL HMGB1-treated group (P<0.01). Conclusions These data suggest that HMGB1 appears to be a potential immunostimulatory signal that induced DC maturation, and HMGB1 stimulation can result in marked up-regulation of IL-12 as well as TNF-α synthesis and release in splenic DCs.