Background The study on normal stem cell markers provides a new way of thinking of that pathogenesis of cancer research and looking for specific markers of cancer stem cells.Importin13 (IPO13) is a novel nucleus-cytoplasm transport receptor protein of importin β family,the study on the biological behavior of IPO13 in ocular tissue and limbal neoplasms is lacking.Objective This study was to investigate the differential expression of IPO13 and p63 in human benign and malignant conjunctiva-cornea neoplasms.Methods The specimens of normal donor limbal and conjunctival tissues (6),conjunciva-cornea papilloma (CCP) (6) and conjunctiva-cornea intraepithelial neoplasia(CCIN) (6)were collected from Xiangya Hospital of Center South University.The expressions of IPO13 and p63 in the corresponding tissue were qualitatively and quantitatively detected using immunochemistry.The A values of IPO13 and p63 positive response were calculated and compared among the 3 types specimens.Results The immunohistostaining on frozen sections showed that IPO13 was expressed in nuclei of basal cells of limbal and conjunctival epithelium and the cellular nuclei of basal cells and suprabasal cells of CCP and CCIN epithelium.The A values of IPO13 positive expression were 1687± 1014,3546± 1375 and 7635 ±2854 in the normal limbal specimen,CCP and CCIN,respectively,showing a significant difference among them (F =7.23,P＜0.05),and the A value was higher in the CCP and CCIN than that in the normal limbal tissue (q =4.02,5.13,P＜0.05),and that in the CCIN was significantly elevated in comparison with CCP(q =3.45,P＜0.05).p63 was expressed in nuclei of basal cells and suprabasal of limbal,conjunctival and CCP epithelium,and was expressed in nuclei of the entire CCIN epithelium.The expressions of p63 in the normal conjunctiva-cornea tissue,CCP and CCIN were 2110± 1229,3966± 2129 and 6650± 2136 respectively,with significant difference among the three different specimens (F =6.17,P＜ 0.05),and the A value of p63 positive expression was significantly elevated in the CCP and CCIN compared with normal limbal specimen (q =4.33,5.01,P＜0.05),and that in the CCIN was significantly elevated in comparison with CCP(q=3.83,P＜0.05).Conclusions IPO13 is more specifical in marking the poorly differentiated cells in limbal epithelial proliferative lesions than p63.Compared with the normal limbal specimen,the expressions of IPO13 and p63 in CCP and CCIN specimens gradually upregulat,which suggests that IPO13 and p63 may play a positive regulatory role in conjunctiva-cornea proliferative lesions.The differential expression of IPO13 and p63 is predominant between benign and malignant limbal epithelial proliferative lesions,indicating that IPO13 and p63 may play an important role in regulating the abnormal proliferation and differentiation of limbal epithelial proliferative lesions.

OBJECTIVETo establish the determination method of pulegone in Herba Schizonepetae.

METHODHPLC method was developed for the determination of pulegone in Herba Schizonepetae. The separation was performed on C18 column with MeOH-H2O (80:20) as a mobile phase. The detection wavelength was set at 252 nm.

RESULTThe quantitation of pulegone in Herba Schizonepetae varied from 0.2-0.7 mg x g(-1). The average recovery is 96.2%, RSD 0.81%.

CONCLUSIONThe method is simple, rapid and accurate.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Lamiaceae ; chemistry ; Monoterpenes ; analysis ; Plant Components, Aerial ; chemistry ; Plants, Medicinal ; chemistry ; Reproducibility of Results

OBJECTIVEThe pathogenesis of irritable bowel syndrome (IBS), a functional gastrointestinal disorder, is poorly understood. Treatment by Chinese medicine and pharmacy (CMP) is superior to treatment by Western medicine (WM). Therefore, we aimed to analyze Chinese medical experts' experiences in treating IBS by data mining.

METHODSBy retrieving related literatures from 1994 to 2012 from CNKI, we chose proved cases and screened effective medical records to establish a database using structural electronic medical case collecting system and data mining. The thinking ways of modern physicians in diagnosis and treatment of IBS were analyzed.

RESULTSTotally 116 medical cases from 46 Chinese medical experts were recruited and analyzed. Results showed that Gan depression and Pi deficiency were most basic syndrome elements. In addition, cold dampness, damp heat, yang deficiency, and Shen deficiency were main elements for the development of diarrhea predominant IBS (IBS-D). Qi stagnation, yin deficiency, qi deficiency, and blood stasis were main elements for the development of constipation predominant IBS (IBS-C). Soothing Gan-qi and strengthening Pi was the most important treatment method. White peony root, prepared atractylodes, prepared licorice, tangerine peel, poria, bupleurum, Radix sileris, lanceolata, and Radix aucklandiae were the most common Chinese herbs used in treating IBS.

CONCLUSIONS(1) Gan depression and Pi deficiency was the main pathogenesis for IBS. Diagnosis and treatment of IBS should be performed clinically according to its development. Tongxie Yao-fang should be used as the basic prescription with modification according to syndrome differentiation. (2) Data mining will have a wide application prospect in studying medical cases.

Data Mining ; Humans ; Irritable Bowel Syndrome ; diagnosis ; drug therapy ; Medicine, Chinese Traditional ; methods ; Phytotherapy ; methods

0.05).(4) Toxicity:Grade Ⅲ-Ⅳ myelosuppression was 60%(18/30)in Tp group,26%(8/31)in TC group and 30%(10/33)in PC group.The TP regimen had the greatest hematologic toxicity(P0.05). Conclusions As first line chemotherapy in epithelial ovarian cancer,TP regimen comparable to the standard chemotherapy regimen.

Adenosine Triphosphate ; Atropine ; Child, Preschool ; Echocardiography, Stress ; methods ; Female ; Humans ; Infant ; Male ; Mucocutaneous Lymph Node Syndrome ; complications ; diagnostic imaging ; Myocardial Ischemia ; diagnostic imaging ; etiology

Multi-template molecularly imprinted solid phase extraction not only has the advantages of high selectivity, large adsorption capacity, easy preparation, reuse and low environmental pollution, but also can realize the enrichment and separation of many kinds of compounds. It has attracted wide attention in the extraction and separation of traditional Chinese medicine components. This study summarizes the latest development of multi-template molecularly imprinted solid phase extraction. At the same time, based on the classification of active components of traditional Chinese medicine (flavonoids, alkaloids, phenylpropanol, terpenes, etc.), the latest application of multi-template molecular imprinting solid phase extraction in multi-component separation of traditional Chinese medicine was reviewed, with a view to better application of multi-template molecularly imprinted polymer in active multi-component extraction and separation of traditional Chinese medicine and provide reference for the material basic research of the efficacy of traditional Chinese medicine.

OBJECTIVETo observe the effects of early goal-directed fluid therapy with hydroxyethyl starch 130/0.4 on intra-abdominal hypertension (IAH), multiple organ dysfunction and fluid balance in severe acute pancreatitis (SAP) patients.

METHODSAccording to the criteria of selection and exclusion, 120 SAP patients within 72 hours after the symptom occurred from 4 study sites were recruited. They were given standard medication according to "the guideline of diagnosis and treatment of SAP in China" in SICU or PICU. The patients were randomly divided into two groups with crystalloid (control group) and colloid plus crystalloid resuscitation (research group). The objective of fluid therapy was to keep steady hemodynamics for 8 days. IAP was measured three times daily by means of urinary bladder transduction. Function of liver, renal and lung were detected daily. APACHE II score and fluid balance were calculated daily.

RESULTSTotal 120 cases were recruited into research group (n = 59) and control group (n = 61). The demography and baseline data were comparable. IAP was lower in research group than that in control group at day 4 and day 5 (P < 0.05). There was no significant difference in APACHE II scores between two groups pre- and after admission. The decline of daily IAP to baseline (DeltaIAP) in research group was significantly higher than in research group from day 2 to day 8(P < 0.05), whilst the decline of daily APACHE II score to baseline (DeltaAPACHE II score) in research group were significantly higher from day 4 to day 8 (P < 0.05). Negative fluid balance emerged much earlier in the research group (P = 0.036). Percentage of patients with negative fluid balance within 8 days was significantly higher in research group than that in control group (94.9% vs. 62.3%). The amount of positive fluid balance was significantly lower in research group (P = 0.039). IAP correlated significantly with APACHE II score (r(2) = 0.322, P = 0.000). PaO2/FiO2 was significantly higer in research group at day 4 and day 8.

CONCLUSIONSIt is very important to pay close attention to IAP in early fluid therapy of SAP patients. Early goal-directed fluid therapy with HES130/0.4 shortens the duration of positive fluid balance, decreases the amount of positive fluid balance, reduces APACHE II score, relieves IAH, and improves PaO2/FiO2.

Fluid Therapy ; Goals ; Humans ; Intra-Abdominal Hypertension ; Multiple Organ Failure ; Pancreatitis

BACKGROUNDIt has already been recognized that psychosocial stress evokes asthma exacerbation; however, the mechanism of how stress gets inside the body is not clear. This study aimed to observe the impact of psychosocial stress on airway inflammation and its mechanism in the ovalbumin-induced asthmatic mice combined with social disruption stress.

METHODSThirty-six male BALB/c mice were randomly divided into: control group, asthma group (ovalbumin-induced), asthma plus social disruption stress group (SDR), and SDR group. The open field video tracking system was used to assess animal behaviors. The invasive pulmonary resistance (RL) and dynamic lung compliance (cdyn) test system from Buxco was applied to detect pulmonary function. The enzyme-linked immunosorbent assay (ELISA) was utilized to determine OVA-IgE, T-helper type 2 (Th2) cytokines (IL-4, IL-5, IL-13) and corticosterone in mouse serum, the Th2 cytokines (IL-4, IL-5, IL-13, IL-6, TNF-α) in bronchoalveolar lavage fluid (BALF), and IL-6 and TNF-α levels in the supernatant of splenocytes cultured in vitro. Hematoxylin-eosin (H&E) staining was used to assess airway inflammation in lung histology. The cell count kit-8 assay (CCK-8) was applied to evaluate the inhibitory effect of corticosterone on splenocyte proliferation induced by lipopolysaccharide (LPS). Real time-PCR and Western blotting were utilized to determine glucocorticoid receptor (GR) mRNA and GR protein expression in lungs.

RESULTSThe open field test showed that combined allergen exposure and repeated stress significantly shortened the time the mice spent in the center of the open field (P < 0.01), increased ambulatory activity (P < 0.01) and the count of fecal boli (P < 0.01), but deceased vertical activity (P < 0.01). Results from pulmonary function demonstrated that airway hyperresponsiveness (AHR) was enhanced by psychosocial stress compared with allergy exposure alone. The ELISA results showed that cytokines in serum and BALF were significantly increased (P < 0.05). Moreover, the lung histology showed that infiltrated inflammatory cells were significantly increased in the asthma-SDR group compared with the asthma group (P < 0.05). Interestingly, serum corticosterone was remarkably raised by psychosocial stress (P < 0.05). In addition, the inhibitory effect of corticosterone on IL-6 and TNF-α in LPS-stimulated splenocyte cultures in vitro was diminished in the asthma-SDR group compared to the asthma group. The CCK-8 test revealed that the inhibition effect of corticosterone on splenocyte proliferation induced by LPS was significantly impaired in the SDR and asthma-SDR groups, while no significant effect was observed in the control and asthma groups. Furthermore, expression of GR mRNA and GR protein were significantly reduced in the lung tissues of the asthma-SDR group (P < 0.05).

CONCLUSIONSSocial disruption stress can promote anxiety behavior, activate the hypothalamic-pituitary-adrenal (HPA) axis, increase AHR and inflammation, and also impair glucocorticoid sensitivity and its function in a murine model of asthma. The down-regulation of GR expression induced by social disruption stress is in part associated with glucocorticoid insensitivity, which leads to asthma exacerbation.

Animals ; Anxiety ; etiology ; Asthma ; etiology ; Bronchial Hyperreactivity ; etiology ; Corticosterone ; blood ; Cytokines ; biosynthesis ; Disease Models, Animal ; Lung ; pathology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Receptors, Glucocorticoid ; analysis ; physiology ; Stress, Psychological ; complications

This study was purposed to investigate whether phenylhexyl isothiocyanate (PHI) can reduce p16 gene methylation level or not. The myeloma U226 cell line was cultured with PHI of 0, 5, 10 micromol/L for 72 hours, then DNA was extracted. Hydrosulfite was used to treat the genome DNA of healthy adult, PCR amplification was carried out by using this DNA as template. The gene chip detecting methylation changes of 3 CpG in promoter region of p16 gene was constructed by designing a pair of probes which contain one methylated and one unmethylated probes. This pair of probes was used to detect 3 consecutive sites of CpG island in p16 gene. The standard curve was constructed by using gene chip after the methylated and unmethylated DNA were mixed at different ratio. Then treated samples of U266 cells were dotted on gene chip, obtained results were compared with standard curve to get the quantitative results. The results indicated that the probes on chip had excellent reproducible ability and precision, the methylation level of p16 gene in U266 cells treated with 0, 5 and 10 micromol/L of PHI was determined as 78.2%, 61.7% and 54.8%, respectively. It is concluded that the PHI can reduce the methylation level of p16 gene in U266 cells.
Cell Line, Tumor ; CpG Islands ; DNA Methylation ; Down-Regulation ; Genes, p16 ; Humans ; Isothiocyanates ; pharmacology ; Oligonucleotide Array Sequence Analysis

OBJECTIVETo report a Chinese Charcot-Marie-Tooth disease type 2 (CMT2) family.

METHODSAll the members in the family were studied clinically,and 6 patients were studied electrophysiologically. Sural nerve biopsy was performed in the proband. PMP22 gene duplications were detected by highly polymorphic short tandem repeat. Point mutation analysis of PMP22, MPZ and NEFL gene was screened by PCR-SSCP combined with DNA direct sequencing. A genome-wide screening was carried out to the family.

RESULTExcept 2 who had weakness and atrophy in both proximal and distal muscles of the lower limbs, all patients presented muscle wasting and a predominating weakness of distal parts of the lower limbs, and mild to moderate sensory impairments. In 6 patients who were subjected to elctrophysiological examinations, median-nerve conduction velocity (NCV) of the median nerve was normal. Electromyograms (EMGs) revealed signs of denervation with large motor unit potentials, fibrillation potentials and positive sharp waves. Sural nerve biopsy of the proband confirmed the presence of axonal neuropathy with an important loss of large myelinating fibers and a large number of clusters with mostly thinly myelinated axons. PMP22, MPZ and NEFL gene mutations were not found. The results of genome-wide screening revealed a linkage of CMT2 to a locus at chromosome 12q24.

CONCLUSIONThe results are consistent with the diagnosis of CMT2. This family represents a rare genetic type of CMT2 which can be designated as CMT2L.

Adolescent ; Adult ; Asian Continental Ancestry Group ; Charcot-Marie-Tooth Disease ; genetics ; pathology ; Chromosomes, Human, Pair 12 ; genetics ; Electromyography ; Female ; Humans ; Male ; Pedigree