Adult ; China ; Humans ; Male ; Neisseria meningitidis, Serogroup C ; drug effects ; genetics ; isolation & purification

Objective To evaluate the feasibility, methods and effectiveness of using a mechani-cal stapler for choledochojejunostomy.Methods The authors have operated on 118 patients in the management of carcinoma of head of pancreas, or periampullary tumor, or cholelithiasis.In the opera-tion, the bilio-enteric end-to-side, or end-to-end and side-to-side anastomosis was made by a circular stapler device, and then a Roux-en-Y or Brown's loop was formed for the preeedure.Results All the surgery of using stapler was done successfully.No postoperative complications such as stomal leak, bleeding and narrow were found.Meanwhile, no harmful consequences were observed through long-time follow-up.Conclusion Using mechanical stapler for bilio-intestinal anastomosis is time-saving, simple and reliable.It can be a choice for some diseases.

Objective To observe the effect of inducible costimulator(ICOS) costimulation pathway blockade in rat limb allografts acute rejection by RNA interference. Methods Twenty-seven cases of modified hind llmb allotransplantation were performed from Wistar to SD rats. The rats were divided into 3 gronps(each n=9): the rejection group not given a special disposal; the control group, consisting of SD rats that received injection of pSilencer 4.1 and Sofast complex by vein post transplantation; and the interference group that received injection of pSilencer 4.1-ICOSshRNA and Sofast complex. On the eighth day posttransplantation, 3 rats were killed to study the pathological changes in each group. The expressions of ICOS gene in vivo were detected by flow cytometry and RT-PCR. The mixed lymphocyte reaction (MLR) was performed and eytokines in blood were measured by ELISA. The rest rats were used to record limb survival time. Results The mean survival time in rats of the rejection and the control groups were(11.34±1.21) and (11.14±1.32) days respectively. In the interference group, the mean survival time of limb allografts was (16.85±1.73) days(P<0.05). The rats in the rejection and the control groups experienced moderate to serious acute rejections with skin epidermal necrosis, a large quantity of lymphocyte infdtration, muscle cell necrosis and interstitial edema, while the pathological changes in rats of the interference group were mild. The splenocyte ICOS mRNA expression level in the interference group(18.75%) was significantly lower than that of the rejection group(100%) and the control group(98.51%). ICOS cell surface expression level as judged by the fluorescence intensity was 45.59±12.87 in the interference group, 103.72±21.76 in the rejection group, and 93.47±29.55 in the control group(F=6.89, P<0.05). In stimulation assays, a one-way mixed lymphocyte reaction stimulation index(SI), with spleen cells from Wistar and Lewis rats, respectively, the rejection group (5.26±0.42,5.18±0.29) and the control group (5.37±0.27,4.93±0.44) had significantly greater reactions than the interference group(2.37±0.35, 4.87±0.36), respectivily(F=7.29, P<0.05; F=6.19, P0.05). In the IFN-γ and IL-4 expression assays, reactions of the interference group (230.17±38.47,160.32±59.13) were lower than those of the rejection group(490.73±51.48,230.67±45.21) and the control group(480.15±43.96, 240.53± 47.36), (F=7.23,6.75, all P<0.01). Conclusions In vivo transfection of pSilencer 4.1-ICOS shRNA interference plasmid can effectively block T-cell co-stimulation pathway, suppress acute rejection, and prolong limb allografts survival.

To study the effects and possible mechanism of Vitamin K(2) (VK(2)) in the treatment of MDS-JSN04 cells, the changes of morphologic features of MDS-JSN04 cells were investigated by cytomorphology, the apoptosis of MDS-JSN04 cells was observed by transmission electron microscope; cellular proliferation was determined by the MTT assay; cell apoptosis, cell cycle shift and expression of myeloid-specific differentiation antigen (CD11b, CD13) were analyzed by flow cytometry (FCM). The expression of apoptosis-related genes bcl-2, survivin and bax were detected by retrotranscriptase polymerase chain reaction (RT-PCR); the activity of caspase-3 was determined by chemiluminescence assay. The results showed that the typical apoptotic morphological features appeared in cells treated with VK(2) for 72 hours; VK(2) induced apoptosis of MDS-JSN04 cells and in a dose-and-time-dependent manner, G(0)/G(1) cell arrest and significantly down-regulated the expression of bcl-2 and survivin, but had no effect on the expression of bax; the activity of caspase-3 significantly increased. It is concluded that VK(2) induces apoptosis of MDS-JSN04 cells through activating caspase-3 pathways and the apoptosis-related genes bcl-2, survivin may play an important role in this process.
Apoptosis ; drug effects ; CD11b Antigen ; analysis ; CD13 Antigens ; analysis ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; Humans ; Inhibitor of Apoptosis Proteins ; Luminescent Measurements ; methods ; Microscopy, Electron, Transmission ; Microtubule-Associated Proteins ; genetics ; Myelodysplastic Syndromes ; genetics ; metabolism ; pathology ; Neoplasm Proteins ; genetics ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Vitamin K 2 ; pharmacology ; bcl-2-Associated X Protein ; genetics

Objective To investigate the establishment,operation and performance of external quality assessment(EQA) system for microbial morphology and detection of special drug-resistance in clinical laboratory,and explore the value of the developed system in clinical application.Methods The pictures of known bacteria and fungi colony,gram staining and acid-fast staining from clinical microbiology were distributed to the participating laboratories in Gansu province twice a year at regular intervals.The pictures of standard knowledge points from CLSI,such as special drug resistance were distributed simultaneously.All the participating laboratories were required to complete the interpretation for the pictures and report their resuhs in a scheduled time.Then the resuhs were summarized and analyzed as 3 modes:complete consistency,general consistency and non-consistency.Results During the 2 years when the EQA system for microbial morphology and detection of special drug-resistance were performed for 24 times,the rate of annual complete consistency increased year by year and reached to 91.3％ in 2015.Conclusion The EQA system based on the examinations of microbial morphology and CLSI standard knowledge points for clinical laboratory may supervise the staff of clinical microbiology laboratories in the hospitals at second grade or above to master the skills of morphological identification and learn CLSI knowledge points,so their professional skills of clinical microbiology could be comprehensively improved.

Objective To analyze the thickness of cornea and corneal epithelium in healthy subjects by optical coherence tomography angiography (OCTA).Methods Totally 100 healthy subjects aged between 20 and 30 years were analyzed by OCTA technique.Using AngioVue OCTA system of retinal imaging mode,and using SSADA algorithm for imaging,the cornea and the corneal epithelium in the central corneal diameter range of 9 mm were measured.The differences of corneal and corneal epithelial thickness in different gender regions were compared.Results In the male and female group,the corneal central total thickness were (559.92 ±33.26) μm and(540.06 ±31.63)μm,and the corneal epithelial thickness were(57.78 ±4.88) μm and(56.88 ±4.57) μm,The total central corneal thickness and central corneal epithelial thickness of the male were greater than those of the female,the difference was statistically significant (t =3.06,2.10;all P ＜ 0.05).The cornea of male was the thickest at S5,S7 and SN9,there were significant differences at S5 and S7 compared with female (t =2.93,2.83;all P ＜ 0.05);The female cornea was the thickest at S5,SN7 and SN9,and the difference was significant at S5 compared with male.The cornea of male subjects was the thinnest at IT,which was statistically significant only at IT5 compared with female subjects in the same area (t =2.02,P ＜ 0.05);The cornea of female subjects was the thinnest at T5,IT7 and T9,which was statistically significant only at T5 and T9 compared with male subjects in the same region (t =2.63,2.20;all P ＜ 0.05);There was significant difference in corneal thickness between male and female at ST (t =3.1 1,2.79,2.33;all P ＜ 0.05).The corneal epithelium was the thickest at IT5,I7,and I9,and the lowest at S5,S7 and S9,and there was no significant difference compared with female in the same region (all P ＞ 0.05).The corneal epithelium of female at the IT5,T7,N9 were the thickest,SN5,S7,S9 were the thinnest;Except for M2 and SN5,there was no significant differences in corneal epithelium between male and female groups (all P ＞ 0.05).Corneal central epithelium accounted for the largest percentage of total corneal thickness,and gradually decreased from inside to outside.Conclusion OCTA can be used to measure the thickness of corneal and corneal epithelial regions.

Objective To investigate image quality and clinical value of dual-source dual energy virtual non-contrast (VNC) CT of the head. MethodsSixty-two patients suspected of cerebrovascular diseases underwent conventional non-contrast (CNC) CT and dual energy CTA examination of the head with dual-source CT. Virtual non-contrast images were reconstructed using dual energy software. The CT values of gray matter, white matter, cerebrospinal fluid, hyperdense hemorrhagic lesion and hypodense ischemic lesion were compared between CNC and VNC images. A four-score scale was used to assess image quality subjectively. Image noise, radiation dosage and detection rate were compared between CNC and VNC images. Paired t test, Wilcoxon signed ranks test and Chi-square test (McNemar test and Kappa test) were used. Results The CT value on CNC and VNC images, were (43. 3 ± 1.5) and (33. 2 ± 1.3) HU for gray matter (t = 46.98, P ＜ 0. 01), (32. 9 ± 1.3) and (28.8 ± 1.6) HU for white matter(t = 16. 28, P ＜0.01), (9.0 ± 1.4) and (5.3 ± 1.9) HU for cerebrospinal fluid (t=12.41, P＜0.01),(62.8 ±10.0) and (51.3 ± 11.5) HU for hyperdense lesion (Z = -4.37, P ＜ 0.01), (20.7 ±4.7) and (18.0 ±6. 9) HU for hypodense lesion (t = 3. 84, P ＜ 0. 01), respectively. VNC images[(1.63 ±0.34) HU]had more noise than CNC images[(0.99±0.18) HU](Z= -6.41, P＜0.01). VNC [(0. 53 ± 0. 08) mSv]had less effective dose than CNC[(1.37 ± 0. 23) mSy](Z= - 6. 45, P ＜ 0. 01).In subjective assessment, VNC images had more noise (2. 7 ± 0. 5 for VNC and 3.9 ± 0. 3 for CNC,Z = -6. 84, P ＜ 0. 01) and skull base-related artifacts (2. 4 ± 0. 9 for VNC and 3.7 ± 0. 5 for CNC,Z = -6. 15, P ＜0. 01) than CNC images. The gray/white matter contrast (1.3 ± 0. 5 for VNC and 3.3 ±0. 6 for CNC, Z = - 7. 01, P ＜ 0. 01), hyperdense lesion display (3.0 ± 0. 4 for VNC and 4. 0 ± 0. 0 for CNC,Z = -4. 52, P ＜ 0. 01) and hypodense lesion display (3.2 ± 0. 8 for VNC and 3.9 ± 0. 3 for CNC,Z= -3. 12, P ＜0. 01) on VNC images were lower than those on CNC images. In per-patient analysis,29 cases of hyperdense lesion (hemorrhage) were found on VNC images without misdiagnosis. The sensitivity, specificity, positive predictive value and negative predictive value were all 100. 0% (29/29,33/33, 29/29, 33/33). VNC images had the same detection rate of hyperdense lesions as CNC images (P ＞0. 05, Kappa = 1. 000) at per-patient level. Twenty-two patients with hypodense ischemic lesions were found on VNC images with one false positive case and two false negative cases. The sensitivity,specificity, positive predictive value and negative predictive value were 91.3% (21/23), 97.4%(38/39), 95.5% (21/22) and 95.0% (38/40) respectively. No statistical difference was found in detecting hypodense lesions between VNC and CNC images (χ2 = 0. 00, P ＞ 0. 05, Kappa = 0. 895). In per-lesion analysis, 53 hemorrhage lesions were found on VNC images with false negative results of four lesions and no false positive result. The sensitivity, specificity, positive predictive value and negative predictive value were 93.0% (53/57), 100. 0% (38/38), 100. 0% (53/53) and 90. 5% (38/42)respectively. There was no significant difference in detection rate of hyperdense lesion between VNC and CNC images (χ2 =2. 25, P ＞0. 05, Kappa =0. 914). Thirty-eight hypodense lesions were found on VNC images with 2 false positive lesions and 13 false negative lesions. The sensitivity, specificity, positive predictive value and negative predictive value were 73.5% (36/49), 96.4% (53/55), 94. 7% (36/38)and 80. 3% (53/66) respectively. The detection rate of hypodense lesion on VNC images was lower than that on CNC images (χ2 = 6. 67 ,P ＜ 0.01, Kappa = 0. 707). Conclusion Compared with CNC images,head VNC images have reduced image quality and radiation dosage. VNC images can replace CNC images potentially in detecting intracranial hemorrhage and provide information for ischemic cerebrovascular diseases to some extent.

In eukaryotes protein phosphorytion is a key event. By reversible protein phosphorylation eukaryotes control many cellular processes including signal transduction, gene expression, the cell cycle etc. Phosphoproteomics involves identification of phosphoproteins and phosphopeptides, localization of the exact residues that are phosphorylated and quantitation of phosphorylation. Because protein phosphorylation is a dynamic process, and it is present at low abundance within cells, and the phosphorylated sites on proteins might vary, and mass spectrometry (MS) signals from phosphopeptides are usually suppressed etc., so phosphoprotein analysis have more difficulties than nonphosphoprotein. In this article, we outline several analysis techniques for separation, identification and quantitation of phosphorylated proteins and peptides, and discuss the progress in these techniques. At present, MS is still an essential core identification technology for phosphoproteomic studies, To search better enrichment strategies are the main challenges in this rapidly evolving field. A major goal of quantitative proteomics is precise quantification and identification of proteins in complex mixtures. A common method for quantitative proteome analysis is the stable isotope labeling method. Today there is no single method that supersedes all others techniques for Phosphoproteomic studies. With continued development of sample preparation techniques and instrumentation, it should be possible to perform a global analysis of protein phosphorylation.
Animals ; Humans ; Mass Spectrometry ; Phosphoproteins ; analysis ; Phosphorylation ; Proteomics ; methods

OBJECTIVETo investigate the protective effect of Qihuang Mingmu capsule (QHMM) on retina of diabetic mice and its impact on VEGF expression.

METHODForty KK/Upj-Ay mice were randomly divided into the model group and high, middle and low dose QHMM (8.32, 4.16, 2.08 g x kg(-1)) groups. Additional 10 C57BL/6 mice were selected as the control group. Mice were orally administered for three months. Their general appearance, fasting blood-glucose (FBG) and glycosylated hemoglobin (HbA1c) were observed. Pathological changes of retina were observed by light microscope and electron microscope. The expressions of vascular endothelial growth factor (VEGF), growth factor receptors-1 (Flt-1) and growth factor receptors-2 (Flk-1) were examined by Real-time PCR (qPCR) and Western blot.

RESULTQHMM could ameliorate the symptoms of diabetic mice to varying degrees, decrease FBG and HbA1c, alleviate pathological lesions of retina and decrease the expressions of VEGF, Flt-1, Flk-1 mRNA and protein.

CONCLUSIONQHMM has the protective effect on diabetic retinopathy of mice by inhibiting the expressions of VEGF, Flt-1 and Flk-1 and intervening VEGF-VEGFR signal transduction pathway.

Animals ; Capsules ; administration & dosage ; Diabetic Retinopathy ; drug therapy ; genetics ; metabolism ; prevention & control ; Drugs, Chinese Herbal ; administration & dosage ; Gene Expression ; drug effects ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Protective Agents ; administration & dosage ; Retinal Diseases ; drug therapy ; genetics ; metabolism ; prevention & control ; Vascular Endothelial Growth Factor A ; genetics ; metabolism

OBJECTIVETo in vitro study the inhibition effect and possible mechanism of As2S3 nanoparticles (As2S3 nano) on human MDS cell line MUTZ-1 and to compare with that of traditional As2S3.

METHODMUTZ-1 cells were treated with As2S3 nano and traditional regular-sized particles (TRSP) at different concentrations. The cell growth inhibition rate was determined by MTT assay, cell apoptosis by morphology and flow cytometry (FCM), cell cycle by FCM and the activity of caspase-3 by chemiluminescence assay.

RESULTSTreatment of As2S3 nano and TRSP at concentrations of 2, 4, 8 and 16 micromol/L for 48 h could lead to a significant dose-dependent decrease of MUTZ-1 cells and induce apoptosis. The percentages of inhibition were 48.9%, 75.9%, 89.4% and 96.5% in As2S3 nano vs 14.5%, 25.4%, 34.7% and 51.5% in TRSP and apoptosis rates were (12.9 +/- 1.9)%, (19.2 +/- 2.2)%, (30.1 +/- 2.5)% and (45.9 +/- 2.3)% in As2S3 nano vs (5.3 +/- 1.8%)%, (11.1 +/- 2.6)%, (19.3 +/- 2.3)% and (25.5 +/- 2.5)% in TRSP respectively. There was statistically significant difference in these two groups (P < 0.01). The proportion of cell in G2/M phase and the activity of caspase-3 of MUTZ-1 cells treated with A2S32 nano were significantly higher than those treated with control group and As2S3 TRSP groups (P < 0.01).

CONCLUSIONSAs2S3 nanoparticles and TRSP can inhibit the proliferation of MUTZ-1 cells and induce apoptosis, which maybe through activating caspase-3 pathways and increasing the proportion of G2/M phase. As2S3 nanoparticles can produce a much better antitumor effect than As2S3 TRSP do.

Apoptosis ; drug effects ; Arsenicals ; administration & dosage ; pharmacology ; Caspase 3 ; metabolism ; Cell Cycle ; drug effects ; Cell Line ; Cell Proliferation ; drug effects ; Humans ; Myelodysplastic Syndromes ; metabolism ; pathology ; Nanoparticles