Objective To evaluate the effect of Chinese gentian extracts on the proliferation of,apoptosis and phosphorylation of epidermal growth factor receptor (EGFR) in HaCaT cells induced by epidermal growth factor (EGF).Methods Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation of HaCaT cells pretreated with EGF of 20 μg/L for 24 hours followed by 24 hours of treatment with various concentrations of Chinese gentian extracts.Flow cytometry was carried out to detect apoptosis in HaCaT cells pretreated with EGF of 20 μg/L for 24 hours followed by 4 hours of treatment with different concentrations of Chinese gentian extracts.Western blot was conducted to measure the level of phosphorylated EGFR in HaCaT cells treated with different concentrations of Chinese gentian extracts for 24 hours followed by treatment with EGF of 20 μg/L for 10 minutes.Results Chinese gentian extracts inhibited the proliferation (r =-0.991,P ＜ 0.01),but promoted the apoptosis (r =0.996,P ＜ 0.05) of HaCaT cells induced by EGF in a dose-dependent manner.At the same time,the extracts suppressed the phosphorylation of EGFR in HaCaT cells induced by EGF,and the suppressing effect increased with the rise in the concentration of the extracts.Conclusions Chinese gentian may inhibit the proliferation,but promote the apoptosis of keratinocytes by decreasing EGFR phosphorylation and blocking relevant intracellular signaling pathways.

AIM: To investigate the expression of osteopontin (OPN) and CD44 in human renal proximal tubular epithelial cells stimulated by human serum albumin (HSA). METHODS: Proximal tubular epithelial HK-2 cells were stimulated by HSA at different concentrations for different time, then OPN mRNA production was detected by RT-PCR, and OPN protein was detected by Western blotting. The expression of OPN and CD44 in HK-2 cells after stimulation for 24 h or 48 h were detected by immunofluorescence with confocal laser scanning microscope. RESULTS: Osteopontin mRNA in HK-2 cells showed a highest expression at 3 h and 48 h after HSA stimulation. However, the expression of OPN protein in HK-2 cells reached the maximum at 24 h after HSA stimulation. OPN mRNA and protein showed a strong dose-dependence relation with the concentration of HSA. HSA also stimulated HK-2 cells to express CD44 protein, the fluorescence of CD44 was most prominent at 48 h after HSA stimulation. CONCLUSION: HSA stimulates human renal proximal tubular epithelial cells to express OPN and CD44.

ObjectiveTo investigate MR imaging features of endolymphatic sac and vestibular aqueduct in patients with large vestibular aqueduct syndrome (LVAS) and its correlation with hearing loss.MethodsMR imaging findings of LVAS were analyzed in 31 cases (62 ears) retrospectively.MR imaging features were grouped into 4 types.In the first type,the signals of endolymphatic and vesitibular aqueduct were hypointense without any hyperintense area.In the second type,the signals of endolymphatic sac and vestibular were hyperintense which were confined within vestibular fissure.In the third type,the area from vestibular aqueduct backward out of the edge of the petrous bone was hyperintense,but its lower boundary was above posterior semicircular.In the fourth type the area which was hyperintense was below the posterior semicircular.To avoid errors in visual inspection,the hyperintense and hypointense area of endolymphatic and the signal intensity of vestibular aqueduct and cerebrospinal fluid (CSF)were measured.The differences of signal intensity among the vestibular endolymphatic sac between the high-signal areas and lowsignal areas were compared with paired t-test.The correlation of the endolymphatic sac MRI classification and degree of hearing losswasanalyzedby correctedChi-squaretestandSpearmancorrelation analysis.ResultTen ears belonged to type Ⅰ (moderate hearing loss in 1 ear,severe in 4 ears,profound in 5 ears),17 ears belonged to type Ⅱ ( moderate hearing loss in 1 ear; severe in 5 ears,profound in 11 ears),23 ears to type Ⅲ (moderate hearing loss in 3 ear,severe in 5 ears,profound in 15 ears) and 12 ears belonged to Ⅳ(mild hearing loss in 1 ear,moderate in 1 ear,severe 3 ear,profound in 7 ears).The boundary between hyperintense and hypointense area was clear,and the signal intensity ratios was 2.02 ± 0.06.The signal ratios of hyperintense and hypointense area to vestibular and CSF were 0.95 ±0.12,0.49 ±0.10,0.99 ± 0.08 respecitively.So there was statistical significant difference between hyperintense and hypointense area ( t =- 24.966,P ＜ 0.05 ),but there was no statistical significant difference between hyperintense area and vesitbular( t =-24.966,P ＞ 0.05).There was no difference of hearing loss between different MRI types ( likelihood ratio =5.02,P ＞ 0.05 ).Conclusions Not only endolymphatic sac enlarged but also perilymph herniated into skeletal fissures of vestibular aqueduct in patients with LVAS.The signal intensity of the endolymphatic sac did not show significant correlation with degree of hearing loss.

Objective To explore the characteristics of dipyridamole 201 Tl myocardial perfusion imaging (MPI) SPECT in patients with dilated cardiomyopathy. Methods Thirty patients with dilated cardiomyopathy underwent pharmacological stress 201Tl MPI SPECT after intravenous infusion of dipyridamole (0. 56 mg/kg) for 4 min. The early and delayed SPECT images were acquired respectively at 10 and 240 min after 201Tl injection. The images were analyzed and reported by two or three experienced nuclear medicine physicians. Results All patients were found to have abnormal perfusion patterns at delay imaging, however 90.00% (27/30) were also abnormal at early images. Six patients had reverse redistribution. Conclusion Dipyridamole 201Tl MPI SPECT imaging may be of some value for the assessment of patients with dilated cardiomyopathy.

BackgroundIdiopathic epiretinal membranes(ERMs) is a common eye disease condition that leads to progressive decline of visual acuity. Studying the risk factors relating to this disease will shed light on its pathogenesis and allow opthalmologists to screen the affected individuals among the high-risk population and prepare for prevention and management strategies. ObjectiveThis survey was to investigate the risk factors of idiopathic ERMs in the population undergoing routine health check-ups. MethodsThe clinical data of idiopathic ERMs was obtained from the population of routine health check-ups in Peking Union Medical College Hospital from November 2009 to October 2010. The examination outcomes were compared between the individuals with and without idiopathic ERMs. The demographic and clinical factors associated with idiopathic ERMs were analyzed and assessed using univariate and multivariate logistic regression analyses. Result A total of 27 400 people were included in the survey and idiopathic ERMs were diagnosed in 76 cases. No obvious eye complaint was obtained from the idiopathic ERMs. The number of people affected with idiopathic ERMs was 12 ( 12/11 659 ) in the below 40 years group, 21 (21/4595) in the 51-60 years group and 32 (32/2544) in the over 60 years group. Hypertension, diabetes, diedyslipidemia, renal function insufficiency ,and cataract were found in 42％ ,5％ ,66％ ,6％ and 8％ of the patients, respectively. The univariate logistic regression analyses revealed that significant correlations were found between age,hypertension,hyperlipidemia and history of cataract( P＜0. 01 ). Multivariate regression models showed that the risk of idiopathic ERMs increased in age of 51-60( OR=2. 5,95％ CI:1. 2-5.4,P=0.02) and over 60 years( OR =7.3,95％ CI:3.4-15.6 ,P＜0.01 ) and patients suffering from hyperlipidemia ( OR--2. 1,95％ CI:1. 3-3.5, P＜0. 01 ). ConclusionsOver the age of 50 years and hyperlipidemia are primary risk factors of idiopathic ERM.

OBJECTIVETo investigate the female sexual dysfunction (FSD) in type 2 diabetes patients, by comparing the sexual function between type 2 diabetic women and non-diabetic women with Female Sexual Function Index (FSFI).

METHODS115 type 2 diabetic women and 107 age-matched non-diabetes women were enrolled with similar backgrounds. Their sexual functions were evaluated with FSFI. Metabolic parameters such as body mass index, blood lipid profile, hemoglobin A1C, plasma glucose were also collected.

RESULTSTotal score of FSFI of the type 2 diabetic women were significantly lower than that of the non-diabetic controls (18.27±8.96 vs. 23.02±5.78, P=0.000). Scores of the FSFI domains (desire, arousal, lubrication, orgasm, satisfaction, pain) of the type 2 diabetic group were also lower than those of the control group. According to the FSD criterion (FSFI<25) available in China, the percentage of FSD in the type 2 diabetic group was significantly higher than that of the control group (79.2%vs. 55.0%, P<0.001). These trends seemed more prominent in pre-menopause subgroups. The logistic regression analysis indicated that age and diabetes were independent risk factors of FSD. Body Mass Index (BMI) also had influence in the diabetes group.

CONCLUSIONFindings from this study showed that there are more FDS in Chinese type 2 diabetic women than in their non-diabetic counterparts, especially in pre-menopause participants.

Adult ; Asian Continental Ancestry Group ; Diabetes Mellitus, Type 2 ; complications ; Female ; Humans ; Middle Aged ; Sexual Dysfunction, Physiological ; etiology

OBJECTIVETo detect the expression of dihydropyrimidine dehydrogenase (DPD) in pancreatic ductal adenocarcinoma (PDAC), and to investigate the relationships between DPD expression and the prognosis of PDAC.

METHODSImmunohistochemistry and tissue microarray techniques were used to exam the expression of DPD in the cancerous tissue in 156 patients admitted from January 2005 to December 2009, including 89 males and 67 females, with the age ranging from 35 to 81 years. The median age was 55 years.

RESULTSWith the positive rate of DPD 55.1%, the expression of DPD was correlated with the differentiation (P = 0.001), TNM staging of tumor (P = 0.021). No relationship was observed between the vessel invasion (P = 0.265), lymphatic metastasis (P = 0.123), neural invasion (P = 0.598) and DPD expression. In the follow-up 117 patients the overall median survival time was 14.2 months, in 58 cases expressed negative, the median survival time was 20.6 months; 39 cases expressed "+" and "++", the median survival time was 12.3 months; 20 cases expressed "+++", the median survival time was 6.8 months. The expression of DPD in pancreatic cancer was correlated with the prognosis of patients, those with higher expression pattern exhibited shorter survival time (P < 0.05). Univariate survival analysis revealed that DPD expression, TNM staging, lymphatic metastasis and neural invasion were factors related to prognosis (P < 0.05), while differentiation levels and vessel invasion were not. Multivariate survival analysis revealed that DPD expression (P = 0.002), lymphatic metastasis (P = 0.000) were two independent prognostic factors.

CONCLUSIONSThe expression levels of DPD was correlated to differentiation levels of pancreatic cancer and TNM staging; those with higher expression of DPD showed shorter survival time. DPD expression, lymphatic metastasis were independent prognostic factors for pancreatic cancer.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Pancreatic Ductal ; enzymology ; mortality ; Dihydrouracil Dehydrogenase (NADP) ; metabolism ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Staging ; Pancreatic Neoplasms ; enzymology ; pathology ; Prognosis ; Survival Rate

Background Endophthalmitis is a serious,sight-threatening condition.Identifying the causative microorganisms is very important for available treatment of endophthalmitis.Objective This survey was to analyze the spectrum of organisms causing culture-proven endophthalmitis and their sensitivities to commonly antimicrobial agents.Methods Medical data of patients with culture-proven endophthalmitis at Zhongshan Ophthalmic Center from January 2003 through December 2010 were respectively reviewed.The outcomes included intravitreal isolates and antibiotic sensitivities were analyzed.Results Four hundred and sixty-nine strains of organisms were isolated from 447 eyes of 447 patients with infective endophthalmitis,including 22 eyes of polymicrobial infection.In the organisms,gram-positive organisms were 241 (51.4％),fungi were 125 (26.7％) and gram-negative organisms were 103 (22.0％).The most common organisms were Staphylococcus epidermidis in 29.4％,Aspergillus in 7.7％ and Pseudomonas aeruginosa in 5.3％.In this group of infective patients,the most common clinic settings were posttraumatic endophthalmitis (72.7％),and then were postoperative endophthalmitis (10.5％),endogenous endophthalmitis (9.8％) and keratitis (6.9％).Most gram-positive organism and gram-negative organism were sensitive to levofloxacin and cefoperazone.The susceptibility rate of gram-positive organism to chloromycetin was increased in 2007-2010 years compared with 2003-2006 years (x2=5.398,P＜0.05).The susceptibility rate to ciprofloxacin of gram-negative organisms declined (x2 =5.398,P ＜ 0.05),but that to rifampicin increased in the duration of 2007-2010 compared with 2003-2006 (x2 =4.500,P ＜ 0.05).Conclusions Gram-positive organisms are the most commonly causative organisms of endophthalmitis.Most bacterial organisms are sensitive to levofloxacin and cefoperazone.Local data of culturing and susceptibility test offers a guideline for the treatment of infectious endophthalmitis.

OBJECTIVETo investigate the methylation status of LRP15 gene in acute leukemia (AL) patients and its role in the tumorigenesis.

METHODSThe methylation of LRP15 promoter and first exon of bone marrow mononuclear cells in 73 patients with AL, 10 with chronic leukemia (CL), 9 with hematological benign diseases, and 20 healthy transplantation donors was analyzed by using methylation specific polymerase chain reaction. The methylation of LRP15 gene promoter and first exon in COS7, K562, and HL60 cell lines was also assayed.

RESULTSNo LRP15 gene promoter methylation was detected in COS7 cell line. LRP15 gene promoter was methylated in K562 and HL60 cell lines. No deletion of LRP15 gene was detected in all samples. In nearly all French-American-British leukemia subtypes, we found that frequency of LRP15 methylation in adult patients with AL was 71.23% (52/73). There was no detectable methylation in any of the 20 healthy donors and 8 chronic myeloid leukemia patients. The difference in frequency of LRP15 methylation between AL patients and healthy donors or CL patients (10.00%, 1/10) was significant (P < 0.01). Hypermethylation of LRP15 gene was found in 57.14% (16/28) of newly diagnosed AL patients, 83.33% of relapsed AL patients respectively, which was significantly different (P < 0.05). We also demonstrated LRP15 methylation in 55.56% (5/9) adults with benign hematological diseases.

CONCLUSIONSLRP15 methylation changes are common abnormalities in leukemia. LRP15 is postulated to be a tumor suppressor gene.

Acute Disease ; Animals ; Base Sequence ; COS Cells ; Cell Line, Tumor ; Cercopithecus aethiops ; DNA Methylation ; DNA Primers ; Humans ; Leukemia ; genetics ; Neoplasm Proteins ; genetics ; Promoter Regions, Genetic

OBJECTIVETo observe the effects of small interfere RNA (siRNA) targeting the c-myc in combination with 5-fluorouracil (5-Fu) on the growth of Hep-2 cells in vitro and in vivo.

METHODSHep-2 cells transfected with or without c-myc siRNA were treated with 5-Fu for 48 h. C-myc protein levels in Hep-2 cells were detected using the Western blot. The cell cycle was analyzed by flow cytometry. Hep-2 cells were subcutaneously inoculated into the back of BALB/c nude mice to establish the implanted laryngeal squamous carcinoma model. PBS, c-myc siRNA, and 5-Fu, alone or in combinations were administered i.p. The mice were sacrificed after the treatments and the tumor masses were removed to determine the tumor volume and weight. The inhibitory rate was calculated. Expression of c-myc in tumor tissue was detected by immunocytochemistry and cell apoptosis was analyzed by terminal transferase dUTP nick end labeling (TUNEL).

RESULTSThe protein levels of c-myc decreased after transfected with c-myc siRNA. C-myc siRNA-transfected cells showed an increase in the percentage of cells in the GO-G1 phase and a decrease in the percentage of cells in the S phase. When combined with 5-Fu, the results were improved. The tumor growth was faster in the control group and was significantly slower in the c-myc siRNA plus 5-Fu group than that in the c-myc siRNA group or 5-Fu group (P < 0.05). The tumor weight in the c-myc siRNA plus 5-Fu group was significantly smaller than that in the c-myc siRNA or 5-Fu group (P < 0.05). Immunohistochemistry showed that c-myc siRNA inhibited the expression of c-myc in tumor tissues in the c-myc siRNA group and c-myc siRNA plus 5-Fu group (P < 0.05). The number of apoptotic cells in the c-myc siRNA plus 5-Fu group was higher than those in the c-myc siRNA groups (P < 0.05).

CONCLUSIONSC-myc siRNA inhibits the expression of c-myc in Hep-2 cells and in the tumor tissues of nude mice. C-myc siRNA combined with 5-Fu inhibits the growth of implanted laryngeal squamous carcinoma and promotes cell apoptosis. C-myc could become a novel target for the treatment of laryngeal squamous carcinoma.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Female ; Fluorouracil ; pharmacology ; Gene Silencing ; Humans ; Laryngeal Neoplasms ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Proto-Oncogene Proteins c-myc ; genetics ; RNA, Small Interfering ; genetics ; Transfection