Objective To preliminarily understand the genotype characteristics of Toxoplasma gondii in blood of HIV?posi?tive persons in Lincang City,Yunnan Province. Method Two segments of SAG2 gene of T. gondii from blood samples of HIV?positive persons in Lincang City were extracted and amplified by using the nested PCR method and the genotype was identified and compared with the standard strain(Type I)of Toxoplasma gondii. Results Thirty?five SAG2 genes(241 bp)and 35 SAG2 genes(221 bp)of T. gondii were amplified from 170 blood samples of the HIV?positive people,and 4 of each case were selected and digested with enzyme,then 2 aim gene fragments of each case were chosen and compared with the standard strain (Type I)of T. gondii. The digestion of SAG2 gene(241 bp)showed the genotype of the blood samples was Type I or Type II, and the digestion of SAG2 gene(221 bp)confirmed that the genotype was Type I. Conclusion It is preliminarily confirmed that the genotype of T. gondii in blood of HIV?positive persons in Lincang City,Yunnan Province is Type I.

Objective: To explore the mediation effect of p75 neurotrophin receptor (p75NTR) in the effect of brainderived neurotrophic factor precursor (proBDNF) on viability and neurite growth of murine hippocampal neurons.

Objective To analyze the expression of carcinoembryonic antigen (CEA) in various populations and the simultaneous expression of other markers using multiple tumor marker protein chip, and to discuss the possible clinical relevance. Methods A total of 25 076 profiles of multiple tumor marker protein chip were collected in our hospital for analysis. The elevation of CEA in various populations and commonly-seen tumors was analyzed, and the combined elevation of CEA and other markers was also analyzed in tumors. Results Elevation of CEA in patients with malignant tumors was significantly more than those in patients with benign lesions and normal controls (P<0. 01), with the highest positive rate of CEA seen in the colorectal cancer (41. 85%), followed by pancreatic cancer (37. 97%) and lung carcinoma (37. 16%). CEA was always accompanied by other markers in tumor patients, with the mostly seen elevated marker being CA125, followed by CA19-9 and CA242. The CEA/CA125 was often seen in pancreatic cancer (74. 26%), ovarian cancer (69. 57%), hepatocellular carcinoma (62.13%), and lung cancer (51. 68%); CEA/CA19-9 was often seen in pancreatic cancer (77.23%) and hepatocellular carcinoma (72.34%); CEA/CA242 was often seen in pancreatic cancer(77. 23%) and colorectal cancer(57. 61%); CEA + CA19-9 + CA242 was usually seen in pancreatic cancer(76. 24%), ovarian cancer(52. 17%), and colorectal cancer (51. 32%); and CEA+CA19-9 + CA242 + CA125 was mostly seen in pancreatic cancer(61. 39%). Conclusion CEA is widely expressed in malignant tumors, but it is not specific for malignant tumors. Single elevation of CEA has high value for diagnosis of colorectal cancer, pancreatic cancer, and lung carcinoma. CEA combined with CA125, CA19-9 or CA242 can help to improve the positive rate for diagnosis of pancreatic cancer, ovarian cancer and colorectal cancer.

Shenkang injection (SKI) is a classic prescription composed of Radix Astragali, rhubarb, Astragalus, Safflower, and Salvia. This treatment was approved by the State Food and Drug Administration of China in 1999 for treatment of chronic kidney diseases based on good efficacy and safety. This study aimed to investigate the protective effect of SKI against high glucose (HG)-induced renal tubular cell senescence and its underlying mechanism. Primary renal proximal tubule epithelial cells were cultured in (1) control medium (control group), medium containing 5 mmol/L glucose; (2) mannitol medium (mannitol group), medium containing 5 mmol/L glucose, and 25 mmol/L mannitol; (3) HG medium (HG group) containing 30 mmol/L glucose; (4) SKI treatment at high (200 mg/L), medium (100 mg/L), or low (50 mg/L) concentration in HG medium (HG + SKI group); or (5) 200 mg/L SKI treatment in control medium (control + SKI group) for 72 h. HG-induced senescent cells showed the emergence of senescence associated heterochromatin foci, up-regulation of P16 and cyclin D1, increased senescence-associated β-galactosidase activity, and elevated expression of membrane decoy receptor 2. SKI treatment potently prevented these changes in a dose-independent manner. SKI treatment prevented HG-induced up-regulation of pro-senescence molecule mammalian target of rapamycin and p66Shc and down-regulation of anti-senescence molecules klotho, sirt1, and peroxisome proliferator-activated receptor-g in renal tubular epithelial cells. SKI may be a novel strategy for protecting against HG-induced renal tubular cell senescence in treatment of diabetic nephropathy.
Animals ; Cells, Cultured ; Cellular Senescence ; drug effects ; Cyclin D1 ; metabolism ; Cyclin-Dependent Kinase Inhibitor p16 ; metabolism ; Diabetic Nephropathies ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Epithelial Cells ; drug effects ; metabolism ; Glucose ; Kidney Tubules, Proximal ; physiopathology ; Male ; Mice ; Mice, Inbred C57BL

The present study was aimed to establish a modified method for culturing mouse renal proximal tubular epithelial cells (TECs). Renal cortex was isolated from mouse kidney and scissored into pieces. TECs were separated by digesting scissored renal cortex in type II collagenase combined with strainer filtration, and then cultured in DMEM. The morphology of TECs was observed under inverted microscopy. The cell proliferative ability was assessed by flow cytometry, and cell viability was analyzed by CCK-8 assay. The purity of TECs was identified by immunofluorescence. Immunofluorescence observation showed that more than 95% cells were epithelial marker CK18 positive and more than 90% cells expressed renal proximal TECs marker proteins, Villin, AQP1, and SGLT2. The cells could be subcultured for about 5 times. The cell proliferative ability declined following the repeated passage. This study introduced a modified efficient method for culturing highly purified mouse renal proximal TECs.

PURPOSEThe intraorbital wooden foreign body is often misdiagnosed or missed on computed tomog- raphy (CT) scan, due to the invisible or unclear images. The residual foreign bodies often occur during surgical removal. The clinical manifestations, imaging features and treatment of intraorbital wooden foreign bodies were discussed in this study.

METHODWe retrospectively analyzed 14 cases of intraorbital wooden foreign bodies managed at our hospital between January 2007 and May 2015. All patients underwent orbital CT examination before surgery, and surgery was performed under general anesthesia with orbital wound debridement and suture, as well as exploration and removal of wooden foreign bodies.

RESULTSAt first, 11 cases underwent removal of foreign bodies, including 1 case with incomplete removal and then receiving a secondary surgery. Foreign bodies were not found in three cases with preoperative misdiagnosis and orbital MRI found residual foreign bodies in the orbit. Operations were performed via primary wound approach in eight cases, conjunctival approach in two cases, and anterior orbitotomy in four cases. Postoperatively, one case was complicated with eye injuries, three cases with ocular muscle injuries, eight cases with visual loss, and eight cases with orbital abscess. The length of foreign bodies ranged from 1.8 cm to 11.0 cm. The maximum of four foreign bodies were removed at the same time.

CONCLUSIONBecause the imaging of orbital wooden foreign bodies is complex and varied, MRI should be combined when they are invisible on CT scan. At the same time injuries trajectory and clinical mani- festations of patients should be taken into account. Surgical exploration should be extensive and thor- ough, and foreign bodies and orbital abscess must be cleared.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Eye Foreign Bodies ; diagnostic imaging ; surgery ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Wood

OBJECTIVE: To elucidate the effects of lipopolysaccharide (LPS), phospholipase A(2) (PLA(2)) and oxygen free radical (OFR) on proton transmembrane translocation and H(+)-ATPase. METHODS: The normal rats were sacrificed for preparetion liver mitochondria and submitochondrial particles for experiments in vitro. Submitochondrial particles were incubated with LPS (100 &mgr;g/mL), PLA(2) (10 u/mL) and FeSO(4)/Vit C (30/90 &mgr;mol/L) at 30 degrees C for 30 min. The proton translocation of submitochondrial particles (SMPs) were assayed with the fluorescent probe ACMA (9-amino-6-chloro-2 methoxya cridine). The mitochondria were incubated with different concentration of LPS, PLA(2) and FeSO(4)/Vit C. The H(+)-ATPase, PLA(2) and malondialdehyde (MDA) were assayed. RESULTS: The fluorescent quenching of ACMA and H(+)-ATPase activity in high dose was significantly decreased after treatment with LPS, PLA(2), FeSO(4)/Vit C (P<0.05). The mitochondrial PLA(2) activity and MDA content were significantly increased after treatment with LPS (P<0.01). CONCLUSIONS: FeSO(4)/Vit C in low dose causes increases H(+)-ATPase activity. LPS, PLA(2), FeSO(4)/Vit C might be the important factors changing H(+)-ATPase and proton translocation across the membrane.

OBJECTIVETo explore the effects of c-met-siRNA on the growth and invasion of hepatocellular carcinoma MHCC97-H cells by pSuppressorRetro/c-met-siRNA recombinant plasmid transfection.

METHODSRecombinant plasmid transfection to Phoenix A cells was constructed using the lipofectin method and then the retrovirals containing c-met-siRNA were used to infect target cells MHCC97-H. In vitro, c-met expression was tested by Western blot. Cell proliferation, motility and invasiveness were studied using MTT, cell migration assay, and cell invasion assay, respectively.

RESULTSThe expression of c-met decreased significantly in MHCC97-H cells, and the most effective site of the target sequence was at 537. The growth, motility and invasiveness of MHCC97-H cells were inhibited.

CONCLUSIONThe results indicate that c-met-siRNA can down-regulate the expression of c-met and inhibit hepatocellular carcinoma cell proliferation, motility and invasiveness.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Line, Tumor ; Humans ; Liver Neoplasms ; pathology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Plasmids ; Proto-Oncogene Proteins c-met ; genetics ; metabolism ; RNA, Small Interfering

Objective: To construct a novel carrier with core-shell structure-inner core of pFGF2-EGFP-loaded TACS coated by hydroxybutyl chitosan (HBC), and to explore its effects on granular fat graft survival. Methods: The core-structured particles (TACS-pFGF2-EGFP) and core-shell-structured particles (HBC@ TACS-pFGF2-EGFP) were prepared to explore the release pattern of pFGF2-EGFP of these particles. The expression of FGF2 protein was detected by Western-Blot in 293T cells transfected with the sustained - release microspheres in vitro. Cell proliferation assay demonstrated that 10μg/ml pFGF2 plasmid could promote 293T cells growth. Eighteen New Zealand white rabbits were used for adipose tissue transplantation experiment. Rabbit left ear was treated as experimental group, 2 ml fat granules and HBC@TACS-pFGF2-EGFP were implanted; rabbit right ear was used as control group, 2 ml fat granules and HBC@TACS-empty plasmids were transplanted. The specimens were harvested at 4, 8, 12 weeks separately after fat transplantation. Gross view, HE staining, and immunohistochemical staining were performed to observe graft survival, biological characteristics, and neovascular density. Results: HBC@TACS-pFGF2-EGFP particles could sustained release Pfgf2 gene in vitro, and successfully express FGF2 protein after transfecting 293T cells. At different time points after transplantation, the volume of adipose tissues was gradually reduced with time. The fat volume and survival rate of adipose tissues in the experimental group were significantly higher than that in the control group (P<0.05). HE staining showed that the arrangement of new adipocytes in the experimental group was more regular than that in the control group. Immunohistochemistry staining showed that the micro vessel density and FGF2 protein expression level in the adipose tissue of the experimental group were higher than those in the control group (P< 0.05). Conclusions HBC@TACS-pFGF2-EGFP particles with sustained release of FGF2 can improve the survival of granule fat transplantation.

Objective: To investigate the effect of adding glutathion(GSH) to tumescent solution on autologous fat graft survival. Methods: 14 male and female New Zealand rabbits were divided into experimental group and control group randomly, 7 in each group. Experimental group: The donor areas of the rabbits were injected with 3 ml of tumescent solution with GSH. Control group: The donor areas of the rabbits were injected with 3 ml regular tumescent solution. DCFH-DA probe was used for fluorescent staining of harvested fat cells. Then stained fat cells were measured for the intracellular reactive oxygen species(ROS)content by fluorescence microplate. The grafts were harvested at 3 months after transplantation and assessed by general observation, volume measurement, wet weight measurement, HE staining for the number of fat cells, and CD34 immunohistochemical staining for the measurement of micro-vascular density. T test was performed by using SPSS 24.0. Results: The intracellular ROS content of harvested fat cells in experimental group was lower than that in control group, and the difference was statistically significant (P<0.05). At 3 months after transplantation, the wet weight of fat grafts in experimental group was (1133.21±87.97) mg and that in control group was (718.79±79.27) mg. The volume of fat grafts in experimental group was (1.00±0.04) ml and that in control group was (0.59±0.03) ml. The number of fat cells in experimental group was (746.6±15.7)/10 high magnification vision and that in control group was (350.1±32.4)/10 high magnification vision. The density in group experimental was (8.1±2.0)/high magnification vision and that in control group was (6.7±2.4)/high magnification vision. The grafts′ volume, wet weight, and the number of fat cells in experimental group were higher than those in control group, and the differences were statistically significant (P<0.05). The difference of micro-vascular density between experimental group and control group had no statistical significance (P>0.05). Conclusions The addition of GSH to tumescent solution optimizes the process of autologous fat harvest, thereby improving the survival of fat graft.