BACKGROUND: Raman spectrum, compared with conventional detection technologies, is a rapid, non-invasive and label-free optical method. Its application has become an issue of concern in biomedical research. However, further studies are warranted to optimize the acquisition condition of Raman spectra from different stem cells. OBJECTIVE: To explore the effect of the wavelength of laser and the groove frequency of gratings to obtain the optimized parameter combination for Raman spectrum collection in human stem cells. METHODS: Using human mesenchymal stem cells as samples, the effects of different laser wavelengths (532, 38,785 nm) and different grating groove frequency (600, 1200, 1800 gr/mm) on Raman spectra were compared respectively. Then the different combinations of the wavelength and groove frequency were used and compared in terms of the spectra resolution and acquisition time, and the best acquisition condition was selected and applied in a comparison study on the Raman spectra from human mesenchymal stem cells and human embryonic stem cells. RESULTS AND CONCLUSION: The wavelengths of lasers and groove frequencies of gratings showed compound impacts on both the spikes at different wavenumbers and the ratio between spikes; the combination of 785 nm and 1200 gr/mm was confirmed to be the best spectrum features for human mesenchymal stem cells. The comparison of Raman spectra from human mesenchymal stem cells and human embryonic stem cells implies that the embryonic stem cells contain higher nucleic acids than the mesenchymal stem cells, while the mesenchymal stem cells appear to contain more proteins and lipids.

Objective To study the mechanisms of extrarenal arterial blood supply of renal malignancy for its interventional therapy.Methods Routine abdominal aortography and selective questionable feeding arteriography were performed in 141 patients with renal malignancy.The characteristics and formation mechanisms of extrarenal arterial blood supply for renal malignancy were analyzed.Results Of the 141 patients,extrarenal arterial blood supply of renal malignancy were found in 51 patients and there were 87 branchs.The breakthrough of renal capsule with malignancy were found in those 51 patients.No extrarena]arterial blood supply of renal malignancy was found in 90 patients,including 50 patients with and 40 patients without the renal capsule breakthrough with malignancy.The emerge of extrarenal arterial blood supply of renal malignancy were significantly different(x~2 = 31.64,P

Adult ; Aged ; Carcinoma, Hepatocellular ; epidemiology ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Liver Neoplasms ; epidemiology ; Male ; Middle Aged ; Nucleosides ; administration & dosage ; therapeutic use

AIM: To investigate the alterations of tear film after sutureless large incision manual cataract extraction (SLIMCE). METHODS: Sixty-eight SLIMCE operation eyes were studied with slit-limp microscope, break- up time (BUT), SchirmmerⅠtest (SⅠt),and fluorescence(FL) to observe the alterations of tear film at different time points in postoperation. Impression cytology and microphoto-analyses technique were also applied to observe the goblet cells at different time points postoperation(7,14,30,60,90 days). RESULTS: Subjective complaint of dry eye within 90 days after the operations were significantly increased compare with preoperations(5-27,23,19,16,13; 2-16,14,8,6,3). The schirmmer Ⅰ test were greatly increased in 14 days postoperation(10.1±4.5;15.0±4.7,13.8±5.7),the mean scores of fluorescence increased (0-17,9,5;0-8,3,1) and the mean break-up time decreased in 30 days post-operation(10.3±2.2;5.5±2.3,7.0±2.4,7.9±2.2) (P<0.05). CONCLUSION: SLIMCE operation have effect on the stability of tear film.

OBJECTIVETo investigate the in vivo functional roles of the La autoantigen (La), the human homologue of the 33-kDa vesicle-associated membrane protein-associated protein (hVAP-33), and the subunit gamma of the human eukaryotic initiation factors 2B (eIF2Bgamma) as co-infection factors supporting chronic infection with hepatitis C virus (HCV).

METHODSSmall interfering (si)RNAs were designed against the HCV internal ribosome entry site (IRES) and transfected into Huh7 cells chronically infected with the HCV pseudovirus (designated as Huh7-HCV cells). The IRES siRNA producing the most effective silencing was selected for further analysis by fluorescence quantitative polymerase chain reaction (qPCR). siRNAs designed against La, hVAP-33, and eIF2Bgamma and the IRES-specific siRNA were then transfected, respectively or in various combinations, into the Huh7-HCV cell line for 48 h. The delta CT values were calculated and used to compare the HCV inhibitive efficacies of the siRNAs in isolation or in combination. Western blotting analysis was used to compare the quantity of core protein expression in each group.

RESULTSThe four gene-specific siRNAs, in isolation or in combination, caused inhibition of HCV replication and gene and protein expressions to varying degrees. The combination of La + IRES siRNAs produced the strongest inhibition of HCV core antigen expression. The combinations of hVAP-33 + IRES siRNAs and eIF2Bgamma + IRES siRNAs produced stronger inhibitions of HCV replication and gene and protein expressions than either hVAP-33 siRNA or eIF2Bgamma siRNA alone.

CONCLUSIONLa, hVAP-33, and eIF2Bgamma act as co-infection factors of HCV chronic infection in vivo. HCV replication and gene and protein expression can be inhibited significantly by RNA interference of these co-infection factors and/or HCV IRES.

Autoantigens ; genetics ; Cell Line ; Eukaryotic Initiation Factor-2B ; genetics ; Hepacivirus ; immunology ; physiology ; Humans ; RNA, Small Interfering ; genetics ; Ribonucleoproteins ; genetics ; Vesicular Transport Proteins ; genetics ; Virus Replication

Aim To investigate the effects of high glu-cose on cholesterol metabolism in renal tubular cells and the intervention of the anthocyanins. Methods HK-2 cells were grown in the DMEM medium supple-mented with 10% FBS and were divided into 5 groups:normal glucose group, high glucose group, mannitol group, C3G group and Cy group. Effect of anthocya-nins on cell viability was detected with MTT, and cho-lesterol accumulation was detected with Amplex Red Cholesterol Assay kit and Filipin staining. Expression of ABCA1 was detected with RT-qPCR and Western Blot. Results In compared with control groups, HG significantly promoted cholesterol mass inside the cell and decreased the cholesterol concentration in the me-dium after treatment for 24 h or 48 h. The levels of mRNA and protein of ABCA1 were detected with RT-qPCR and Western blot, and both were decreased in the presence of HG. Whereas treatment with C3G and Cy markedly attenuated HG-induced cholesterol mass inside the cell by up-regulating the expression of AB-CA1. Conclusions High glucose can reduce the ex-pressions of the ABCA1, and then decrease cholesterol efflux and increase the cholesterol accumulation in HK-2 cells. Anthocyanins can decrease cholesterol accu-mulation by up-regulating the expression of ABCA1.

OBJECTIVETo investigate the progression of visual field loss and to explore the prognosis of glaucomatous optic neuropathy in patients with chronic angle-closure glaucoma (CACG) after their intraocular pressures were well controlled under 21 mmHg.

METHODSForty-seven eyes of 29 patients in the Department of Ophthalmology in PUMC Hospital were included. All the patients had at least two separate tests of visual fields using the 24-2 program of the Humphery Visual Field Analyzer after their intraocular pressure were well controlled under 21 mmHg after sugery. The visual fields of patients were followed routinely for at least 1 year. In addition, all patients were divided into 2 groups according to follow-up period: 1-2 years group and over 2 years group. Visual field scores were calculated with the Advanced Glaucoma Intervention Study (AGIS) method. The visual fields were divided 5 sections and the sensitivity and defect depth of each section were calculated.

RESULTNo statistically significant differences were found in terms of AGIS scores, localized sensitivities and localized defects within the time interval of the observation.

CONCLUSIONGlaucomatous optic neuropathy is not likely to progressively deteriorate in CACG cases once their intraocular pressure are well controlled under 21 mmHg.

Aged ; Female ; Follow-Up Studies ; Glaucoma, Angle-Closure ; physiopathology ; surgery ; Humans ; Intraocular Pressure ; Male ; Middle Aged ; Optic Disk ; physiopathology ; Optic Nerve Diseases ; physiopathology ; Retrospective Studies ; Visual Fields

A liquid chromatographic-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of budesonide in dog plasma. Budesonide and the internal standard triamcinolone acetonide were separated from plasma by alkalinized liquid-liquid extraction with ethyl acetate. Chromatographic separation was performed on a Capcell Pak C18 MG column with the mobile phase consisted of acetonitrile -5 mmol x L(-1) ammonium acetate (60:40, v/v) at a flow-rate of 0.50 mL x min(-1). A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM) of the transitions m/z 489 --> m/z 357 and m/z 493 --> m/z 413 for budesonide and the internal standard, respectively. The linear calibration curves were obtained in the concentration range of 25.0-2000 pg x mL(-1). The lower limit of quantification was 25.0 pg x mL(-1). The intra- and inter-day relative standard deviation over the entire concentration range was less than 15%. The accuracy was in the range of -8.1% to -1.7% in terms of relative error. The method was applied to a pharmacokinetic study of budesonide controlled-release capsules in Beagle dogs. Maximal budesonide plasma level was observed after (3.5 +/- 3.3) h and the Cmax was (786 +/- 498) pg x mL(-1) after a single oral administration of 9 mg budesonide capsules, Cmax was increased to (2142 +/- 1515) pg x mL(-1) after multiple oral administration (9 mg x 5 d) of budesonide capsules. This method was selective and rapid, and the sensitivity was sufficient for the purpose of the pharmacokinetic study of budesonide controlled-release formulation.
Animals ; Anti-Inflammatory Agents ; blood ; pharmacokinetics ; Area Under Curve ; Budesonide ; blood ; pharmacokinetics ; Chromatography, Liquid ; methods ; Delayed-Action Preparations ; Dogs ; Male ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; Tandem Mass Spectrometry ; methods

OBJECTIVETo investigate the relationship between body mass index and lower limb alignment of patients with knee osteoarthritis.

METHODSFrom July 2008 to June 2010, 78 patients who suffered from severe knee osteoarthritis were treated with total knee replacement, including 17 males and 61 females, ranging from 41 to 85 years old, with an average age of 68.1 years. The Chinese average adult body mass index (BMI) classification standard was used to classify the patients into three groups: normal group, BMI<24.0, including 3 males and 11 females, totalized 14 patients (28 knees), with the mean age of (69.5 +/- 4.7); overweight group, 24.0 < or = BMI<28.0, including 4 males and 25 females, totalized 29 patients (58 knees), with the mean age of (66.4 +/- 7.9) years; and obese group, BMI > or = 28.0, including 10 males and 25 females, totalized 35 patients(70 knees), with the mean age of (69.1 +/- 8.3) years. The limb X-ray film measuring system was used to measure the lower limb alignment at the supine and weight-bearing position.

RESULTSThere were no statistical significances between the normal group and the overweight group in the knee varus angle at the supine and weight-bearing position (P>0.05). However, keen varus angle of the overweight group increased compared with the other two groups at the supine position,and the angle of the weight-bearing position increased nearly 2 degrees, but no statistical significance. There was no statistical significance between supine and weight-bearing position in the three groups of BMI, but the knee varus angle at the supine position was more than the weight -bearing position in the three groups.

CONCLUSIONThe knee varus angle of obese patients with osteoarthritis in the weight-bearing position increases obviously. It indicates that obesity is an important factor increasing the angle of lower limb alignment.

Adult ; Aged ; Aged, 80 and over ; Biomechanical Phenomena ; Body Mass Index ; Bone Malalignment ; etiology ; Case-Control Studies ; Female ; Humans ; Male ; Middle Aged ; Obesity ; complications ; Osteoarthritis, Knee ; complications ; Supine Position

OBJECTIVETo investigate the correlation between mutation in EGFR tyrosine kinase domain and tumor response as well as prognosis in advanced stage non-small cell lung cancer (NSCLC) treated with iressa.

METHODSFrom May 2002 to Feb. 2005, iressa was orally administered at a dose of 250 mg once daily for 106 advanced stage NSCLC patients until occurrence of disease progression or intolerable toxicity. Cancer tissue was obtained from these patients, and DNA was extracted for analysis of mutation in exon 18 to 24 of EGFR. Exon 18 to 24 of EGFR were amplified by nest PCR, sequenced and analyzed from both sense and antisence directions.

RESULTSPrimary NSCLC tissue specimens consisted of 25 frozen tissue blocks and 81 paraffin-embedded tumor tissue blocks from 106 consecutive NSCLC patients. Mutation was found to be more frequent in the adenocarcinoma than in the squamous cell carcinoma (35.9% vs 14.3%, P =0.033). Mutation was identified in 32 patients (30.2%). Response rate to iressa was 71.9% in the patients with EGFR mutation versus 13.5% in those without mutation (P <0.01). Compared with the patients without EGFR mutation, those with mutation had longer overall survival (median, 13.45 vs. 5.25 months; P<0.01) and median time to progression (median, 8.35 vs. 3.0 months; P <0.01).

CONCLUSIONEGFR mutation may be positively correlated with the response and survival in advanced stage Chinese NSCLC patient treated with iressa.

Adenocarcinoma ; drug therapy ; genetics ; pathology ; Adolescent ; Adult ; Aged ; Antineoplastic Agents ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; pathology ; Carcinoma, Squamous Cell ; drug therapy ; genetics ; pathology ; Exons ; Female ; Follow-Up Studies ; Humans ; Kaplan-Meier Estimate ; Lung Neoplasms ; drug therapy ; genetics ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Point Mutation ; Prognosis ; Quinazolines ; therapeutic use ; Receptor, Epidermal Growth Factor ; genetics ; Sequence Deletion