Child, Preschool ; Diagnosis, Differential ; Female ; Gonadotropin-Releasing Hormone ; therapeutic use ; Humans ; Hysteroscopy ; Pediatrics ; Puberty, Precocious ; diagnosis ; therapy ; Uterine Hemorrhage ; diagnosis ; etiology ; therapy ; Vaginal Diseases ; diagnosis ; etiology ; therapy

Background Human limbal allograft transplantation or limbal autograft transplantation are the primary approaches to the severe corneal-blindness,but their application in clinic were limited because of the defects of donor material.With the development of tissue engineering technology,transplantation of in vitro cultured limbal epithelial stem cells is being an advanced management.Objective The aim of this work was to expand human limbal epithelial stem cells ex vivo under the guidance of confocal microscope and to lay the foundation for fabricating ex vivo cultured cell sheets.Methods Ten eyes of ten patients were examined with the Heidelberg Retina Tomography Ⅲ Rostock Cornea Module(HRT3-RCM)to elucidate the structure of the human corneoscleral limbus and to correlate limbal epithelial dimensions.According to the analysis of the images of limbal epithelia,the limbal tissues provided by Eye Bank of Henan Eye Institute were cut into suitable explants.Then,this study was conducted to expand limbal epithelial stem cells ex vivo on denuded amniotic membrane.The phenotypes of primary cultured cells were evaluated by morphology and immunofluorescent staining with antibodies for limbal epithelial stem cell markers (p63,cytokeratinl9)and differentiation markers(keratin 3,involucrin).This experimental procedure was approved by the Ethic Committee of Henan Provincial People's Hospital.The written informed consent was obtained from subjects before initiation of any examination.Results The palisade morphology of human limbus was imaged clearly on the laser scanning in vivo confocal microscopy and many hyperreflective cells were observed in palisade basal cells.The cell-island phenomenon was seen in the basement membrane under the laser scanning in vivo confocal microscopy.The oblique sections of limbus showed many papilla-like epithelial columns below the superficial limbal epithelia.Throughout the experiment duration,the epithelial cells grew well with the migration rates from limbal tissue (68.62± 16.94)％ and the migration time(5.83 ±2.04)days,which depended on the tissue freshness.Compared with the second and forth batch of tissue,the migration rates of the third and sixth batch of tissues were significantly higher(P＜0.05),and the migration time was evidently longer in the forth and sixth batch of tissue compared with the first,second,third and fifth batch(P＜0.05).The positively expressing rates in the cultured corneal stem cells were 4.05％ and 36.52％ for p63,26.07％ and 40.55％ for CK19,57.88％ and 40.81％ for K3,64.66％ and 59.19％ for involucrin.Conclusion Human limbal epithelial stem cells can be successfully and purposefully obtained from the limbal tissue based on the guidance confocal miscroscope.The cultured corneal stem cells can grow well on the denuded amniotic membrane

OJECTIVE: To explore the changes of serum IgE and tryptase caused by anaphylactic shock rats and discuss the relation to PMI and preservative environment of corpse and specimen. METHODS: Rats were used for establishing anaphylactic shock models and randomly divided into room temperature group, refrigeration group, frozen group, manual hemolysis group, specimen preservation group. And the control group was also established. The blood samples were collected after rats were sacrificed. The degree of hemolysis was graded according to the color of the upper layer of the serum. The mass concentration of IgE and tryptase in each group was detected by ELISA. RESULTS: The levels of serum IgE and tryptase in anaphylactic shock dead rats were higher than that of the control group. Room temperature and frozen made obviously differences on the levels of serum IgE and tryptase with various PMI. The levels of serum IgE and tryptase in refrigeration group showed relatively stable. The levels of serum tryptase and IgE were elevated with differently increasing hemolysis. The levels of serum IgE and tryptase showed no obvious changes during the specimen kept under different temperature conditions for 25 days. CONCLUSION Serum IgE and tryptase obviously increased in anaphylactic shock rats. However, the levels were influenced by PMI and environmental temperature, especially under the conditions of room temperature and frozen.
Anaphylaxis/blood* ; Animals ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Immunoglobulin E/blood* ; Rats ; Temperature ; Tryptases/blood*

BACKGROUND:Previous research indicated that iron-fluorouracil-phenanthroline complex has good antitumor activity in vitro, which can inhibit the proliferation of human cancer cels. OBJECTIVE:To detect the antitumor activity and toxicity of iron-fluouracil-phenanthroline complex, [Fe(5-Fu)2(Phen)SO4],in vivo. METHODS:A total of 40 Kunming mice were randomly divided into four groups, which were intraperitoneally injected with 72, 102.9, 147, 210 mg/kg [Fe(5-Fu)2(Phen)SO4] and the half lethal dose of the complex was detected. One day after the establishment of mouse S180 sarcoma models, the model mice were randomly divided into eight groups, and administered with the intraperitoneal injection of 15 mg/kg (low dose group), 30 mg/kg (middle dose group), 60 mg/kg (high dose group) [Fe(5-Fu)2(Phen)SO4], normal saline (negative control group), cisplatin (positive control group), 5-fluorouracil, iron-salt and phenanthroline, respectively. The injection was done once a day, lasting for 7 days. The weight of sarcomas, body weight, the main organ coefficient and histopathological changes of the main organs were detected. RESULTS AND CONCLUSION: The half lethal dose of [Fe(5-Fu)2(Phen)SO4] was 103.9 mg/kg. Compared with the negative control group, high dose group, positive control group and 5-fluorouracil could significantly inhibit the growth of the tumor (P< 0.05 orP< 0.01), and the effect of high dose group was the most obvious (P < 0.01). Compared with cisplatin, 60mg/kg [Fe(5-Fu)2(Phen)SO4] had a weaker inhibitory effect on the kidney, but higher inhibitory effect on the liver, spleen and thymus, indicating the complex has a lower nephrotoxicity, but stronger immunotoxicity and hepatotoxicity than cisplatin.

Objective To explore the value of SOD activity in diagnosis of central nervous system leukemia (CNSL )by detec-ting SOD activity of cerebrospinal fluid of patients with CNSL .Methods The cerebrospinal fluid of 55 patients from department of hematology of Nanfang hospital of southern medical university were collected from January 2008 to January 2009 ,in which 30 pa-tients suffered with central nervous system leukemia (CNSL group) ,the other 25 patients suffered with acute leucemia without im-paired central nervous system(control group) .The SOD activity of cerebrospinal fluid was detected by the xanthine oxidase meth-od ,while the routine test ,biochemistry test and cell smear of cerebrospinal fluid was detected .Results There were statistics differ-ence in the level of white cell and protein in cerebrospinal fluid between CNSL and control group (P<0 .05) ,but with no difference in the level of cerebrospinal fluid pressure ,glucose ,chlorine(P>0 .05) .There was statistics difference in the level of SOD activity between CNSL and control group(P<0 .05) .The white cell quantity and the protein level in cerebrospinal fluid had negative corre-lation with the activity of SOD ,(r=0 .871 ,P=0 .000 ;r=0 .518 ,P=0 .003) .The activity of SOD in the cerebrospinal fluid had sta-tistics difference before and after intrathecal chemotherapy (P<0 .05) .The activity of SOD in the cerebrospinal fluid whose under 45 year-old (755 .64 ± 345 .77) ,which was significant lower than that of the paitents whose equal with or above 45 year-old (1 420 .49 ± 307 .69)(P<0 .05) .Conclusion The changes of the SOD activity in the cerebrospinal fluid had relation with central nervous system leukemia ,and the SOD activity might be a auxiliary diagnosis index used in central nervous system leukemia by revi-sing age factor .

Objective to investigate the value of diffusion-weighted echo-planar MR imaging in the diagnosis of head and neck lesions.Methods Fifty-seven patients with 85 head and neck lesions were enrolled in the study,including 22 patients with 22 malignant tumors,13 patients with 13 benign tumors, 13 patients with 17 cystic and liquefactive lesions(including 8 patients with 12 cystic lesions,4 patients with 4 tumor necrosis,1 patients with 1 abcess)and 33 lymph nodes.The lesions were all confirmed by operation and clinical follow up.Echo-planar difffusion-weighted imaging (DWI)was performed with different b values (0,500,and 1,000s?mm~(-2)),and the apparent diffusion coefficients (ADCs)were measured.Results Malignant and benign tumors had different characteristics in DWI with different b values.With the increase of b value,the signal intensity of tumor/spinal cord ratio decreased quickly in DWI in benign tumors,while the signal intensity of tumor/spinal cord ratio remained similar in DWI in malignant tumors.The mean ADC value of'malignant tumors[(0.78?0.24)?10~(-3)mm~2? s~(-1)] was significantly lower than that of benign tumors [(1.48?+0.20)?10~(-3)mm~2?s~(-1)] (t = 8.9,P

Fatal anaphylactic shock is common in forensic practice. However, it is difficult to diagnose for lacking specific pathological and morphologic changes in forensic autopsy. The application of some biochemical indicators is of great significance. This paper reviews the biological characteristics of some biochemical indicators and detection methods. The forensic application, problems and prospects of these indicators are also introduced in details. The stable biochemical indicators, IgE, tryptase and chymase, show great potential and advantages in the identification of fatal anaphylactic shock in forensic medicine.
Anaphylaxis/metabolism* ; Autopsy ; Biomarkers ; Chymases ; Forensic Medicine ; Humans ; Tryptases

OBJECTIVETo evaluate the age-dependent alteration of insulin and ghrelin expression in the pancreatic islets of intrauterine growth-retarded (IUGR) rats during development and to identify the role of ghrelin in insulin resistance induced by IUGR.

METHODSNeonatal SD rats were divided into normal birth weight group (N group) and intrauterine growth-retarded group (I group). Plasma glucose, ghrelin and serum insulin were analyzed at day 1, 3, 7 and 10 and at week 2, 3, 4,8 and 12 after birth.Entire pancreas samples were collected for determination of ghrelin and insulin mRNA. Immunohistochemical double-staining and confocal microscopy was performed on rat pancreas.

RESULTPlasma insulin levels of I group were lower than those of N group at day 1, 3 and 7 (P <0.05). The homeostasis model assessment-insulin resistance index (HOMA-IR) of I group was higher than that of N group at day 1, 3 and 7 (P<0.05). Plasma glucose levels, insulin, HOMA-IR and ghrelin concentrations decreased in an age-dependent manner (F = 4.12 to approximately 125.97, P <0.001) in both groups. Plasma ghrelin level was correlated with HOMA-IR in I group(r=0.553, P=0.000). Pancreatic ghrelin contents, pancreatic ghrelin mRNA expression, the percentage of insulin (+) cells and ghrelin (+) cells in both groups decreased in an age-dependent manner (F = -49.29 to approximately 427.28, P<0.001). There were differences in the above indexes between N and I group (F = -69.98 to approximately 169.22, P<0.05). Insulin secretion was negatively correlated with ghrelin contents in both groups (r=-0.895, P=0.000; r=-0.458, P=0.002). Percentage of insulin(+) cells was negatively correlated with the percentage of ghrelin (+) cells in pancreatic islets in both groups (r=-0.810,P=0.000; r=-0.714, P=0.000). The distributions of ghrelin (+) cells in pancreatic islets of I group were different from those of N group.

CONCLUSIONThe effects of IUGR on weight, plasma ghrelin levels and insulin secretion of pups rats persist after birth and ghrelin may be involved in the process of insulin resistance in IUGR rats.

Animals ; Animals, Newborn ; Female ; Fetal Growth Retardation ; genetics ; metabolism ; Ghrelin ; blood ; metabolism ; Insulin ; blood ; metabolism ; Insulin Resistance ; physiology ; Insulin-Secreting Cells ; metabolism ; Pregnancy ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To explore the clinicopathological features and prognostic significance of human epidermal growth factor receptor 2 (HER2) in gastric carcinoma. Methods Pathological data of 127 patients with gastric carcinoma were retrospectively analyzed. HER2 expressions of all patients were detected by immunohistochemistry (IHC). 119 (93.7 %) patients were undergone R0 dissection and 123 (96.9 %) cases received D2 lymph nodes dissection. 51 (40.2 %) patients received adjuvant chemotherapy. The proportional differences of clinicopathological features for patients between HER2-positive and HER2-negative were compared, including the patients' survival. Results HER2 overexpression rate was 8.7 % (11/127) in gastric carcinoma patients. For the patients with HER2-positive and HER-negative, the lymph node metastasis rates were 100.0 % (11/11) and 81.9 % (95/116), respectively (P= 0.041). The 3-year overall survival (OS) rates for HER2-positive and HER2- negative patients with gastric carcinoma were 32.7 % and 42.9 % (P=0.413), and the 5-year progression free survival (PFS) rates were 27.3 % and 42.2 % (P = 0.354), respectively. Among patients with HER2-negative, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 55.3%, compared with 35.4%for patients with surgery alone (P=0.015), and the 3-year PFS rates were 53.3 % and 35.3 % (P= 0.038), respectively. Among patients with HER2-positive patients, 3-year OS rate for patients with surgery plus adjuvant chemotherapy was 0, compared with 75.0%for patients with surgery alone (P=0.002), and the 3-year PFS rates were 0 and 60.0% (P=0.004). Conclusions HER2 is expressed in gastric carcinoma tissue, related to lymph node metastasis. HER2 status are not correlated with the prognosis for gastric carcinoma patients, however, it is likely to be a predictive marker for adjuvant treatment after surgery for patients with gastric carcinoma.

OBJECTIVETo investigate the effect of mitogen Phorbol 12-myristate 13-Acetate (PMA) on CD3, CD4 and CD8 expression of human T-lymphocytes.

METHODSPeripheral blood mononuclear cells from 37 blood samples stimulated in vitro with PMA at different concentrations (2,5,10,20 and 50 ng/ml for 4 hours) and time (10 ng/ml for 2,4 and 6 hours) were analyzed by 4-color flow cytometry (FCM).

RESULTSUnder different PMA stimulation protocols,significant CD4 down-regulation was observed,which was negatively correlated with intracellular cytokine secretion (r= 0.601,P<.001), except for PMA stimulation at 10 ng/ml for 2-hours which showed no significant intracellular cytokine secretion. The expressions of CD3 and CD8 molecules after PMA activation were not significantly affected as compared with pre-activation. Among CD3 positive T lymphocytes, CD4/CD8 double-negative cells only account for 5.52%.

CONCLUSIONPMA has a significant down-regulation effect on CD4 molecules of Th cells, without altering the CD3 and CD8 expression. For quantitative analysis of Th1/Th2 variation, indirect method such as CD3(+)CD8( ) T cells can be used to define CD4(+) Th cells stimulated by PMA in the future.

CD3 Complex ; blood ; CD4 Antigens ; blood ; CD8 Antigens ; blood ; Child ; Humans ; Interferon-gamma ; biosynthesis ; Interleukin-4 ; biosynthesis ; Lymphocyte Activation ; drug effects ; T-Lymphocytes ; chemistry ; drug effects ; Tetradecanoylphorbol Acetate ; pharmacology ; Th1 Cells ; immunology ; Th2 Cells ; immunology