Objective To analyze the protein expression profile of human glomerular mesangial cells (HMCs) under high glucose and to characterize molecular functions and biological processes. Methods HMCs were divided into high glucose cultured group (30 mmol/L) and normal glucose cultured group (5 mmol/L). The total proteins were extracted after culture for 48 hours. The total proteins of the two groups were separated using two-dimensional fluorescence difference in gel electrophoresis (2-D DIGE) and analyzed using DeCyder 2-D difference analysis software. The differentially expressed proteins were further identified using in-gel digestion with trypsin, of which peptide extracts were prepared for MALDI-TOF-MS analysis. Protein identifications were searched in the NCBI protein database using the Mascot search engine. Results One hundred and forty-seven protein spots whose expression levels were significantly increased or decreased more than 1.5 folds under high glucose were identified. Ninety-six differentially expression protein spots were analyzed by peptide mass fingerprinting and 37 kinds of proteins were identified. The protein spots of phosphatidylethanolamine binding protein 1 (PEBP-1), granulysin,ATP synthase H + transporting mitochondrial FO complex subunit F2 were observed only in high glucose group. The expression of 24 proteins was up-regulated by high glucose, including eosinophil cationic protein, RGS membrane-interacting proteins 16 (MIR16), peptidyl-prolyl cis-trans isomerase, disks large homolog DLG2, breast cancer 2, early onset (BRCA2), Catechol-O-methyltransferase etc. The expression of 5 proteins was down-regulated by high glucose, including O-GlcNAc transferase-interacting protein 106 000 isoform 1, proteasome beta 6 subunit precursor,NEFA-interacting nuclear protein NIP30 etc. Conclusions Expression of 147 proteins in HMCs alters under high glucose. These proteins are involved in the regulation of cytoskeleton, glucose metabolism, cell division, gene transcription, signal transduction, phosphorylation, cell proliferation,apoptosis etc. In-depth analysis of these differentially expressed proteins' function and crosstalk is expected to provide an important experimental basis for clarifying the pathogenesis of diabetic nephropathy.

The effects of Guizhi Fuling capsule and its active ingredient combination within different concentration on SPL proliferate were observed by MTT method. The ratio of CD80/86, CD3CD25 and CD3CD69 was used to evaluate cell activation effects of Guizhi Fuling capsule and its active ingredient combination by FCM. Guizhi Fuling capsule with concentration of 400 mg · L(-1)can promote spleen lymphocyte proliferation, as well as the active ingredient combination, which showed the obvious dose-effect relationship. Compared with control group, the difference has statistical significance (P≤0.01). The result of FCM showed that Guizhi Fuling capsule and its active ingredient combination can promote CD80 and CD86 expression on spleen lymphocyte, and also can increase CD25 and CD69 ratio between spleen CD3+ cells. Compared with control group, the difference has statistical significance (P≤0.01). Thus, Guizhi Fuling capsule and its active ingredient combination may have immune-modulate effects, and the mechanism may have a close relationship with the lymphocyte activation.
Animals ; Capsules ; Drugs, Chinese Herbal ; analysis ; pharmacology ; Immunologic Factors ; pharmacology ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; T-Lymphocytes ; drug effects ; immunology

SUMMARY Pregnancy-related pelvic ring disease brings great suffering to pregnant women, including the separation of the pubic symphysis and sacroiliac joint pain.Hormonal changes leading to ligamentous laxity is the main reason for pregnancy-related pelvic ring disease.In normal pregnant cases, and the physiologic widening at the symphysis is about 3-7 mm.When the widening of the symphysis is more than 10 mm, it may lead to symptoms and need active treatment.Currently the diagnosis of the pubic symphysis separation is based on the clinical symptoms and signs.The treatment of acute pubic symphysis separation bases on conservative therapy, includes bed rest and physical therapy.But when the widening of the symphysis is more than 4 cm, the surgery intervention may be a good treatment.If the conservative treatment is not obviously effective, the surgery consists of plate fixation in the pubic symphysis and sac-roiliac screw fixation.Other indications for the surgical intervention include inadequate reduction, recur-rent diastasis, intractable symptoms, and open rupture.

Objective To review our experience in correction of pectus excavatum.by Nuss procedure without thoracoscopic assistance.Methods 412 patients with pectus excavatum were surgically corrected using Nuss technique without thoracoscopic assistance.There were 329 boys and 83 girls with average age of 7 years and 8 months ( range from 2 years and 8 months to 28 yeats and 5 months).The bars were removed after surgery in 102 patients.Results The operation in all patients had been performed successfully without any severe complications The average time of the operation was 40 minutes.The average blood lose during operation was ( 10 ±2) ml.Conclusion Nuss procedure without thoracoscopic assistance is safety and effective.It could be minimize the trauma and shorter operating time.

Objective To explore prevention and cure effects of the freeze-dried human fibrin glue as the way of the lacrimal duct embolization on xerophthalmia in perimenopausal female rabbit.Methods A total of 72 female rabbits,after anti infection treatment and were cut off third eyelid,were made into perimenopausal xerophthalmia rabbit models.After surgery,all of these rabbits were randomly divided into 6 groups (12 rabbits per group):No treatment group after surgery (group A),PBS prevention group (group B),freeze-dried human fibrin prevention group (group C);no treatment group after modeling (modeling time:Two weeks after surgery,group D),PBS treatment group (group E),freeze-dried human fibrin treatmentgroup (group F).The Schirmer test (SIT),corneal fluorescein (FL) and corneal confocal microscope were performed before and 2 weeks,4 weeks,6 weeks after injection.Results There were statistical differences in FL score and SIT in group A,group B and group C among different time points (F =27.346,10.608;P =0.000,0.001);There were statistical differences between FL scores and SIT among three groups (F =7.579,6.786;P =0.002,0.007);There was significant difference in FL scores and SIT trends among three groups(F =44.897,3.424;P =0.000,0.045).The FL score and SIT of group D,group E and group F were significantly improved after treatment for 2 weeks,4 weeks and 6 weeks,the differences were statistically significant (t =2.906,3.654,4.504;P =0.022,0.017,0.013.t =4.573,5.759,7.231;P =0.032,0.019,0.008);The difference between FL score and SIT in group E and group F was statistically significant after treatment (t =2.776,4.124,5.324;P =0.032,0.026,0.017.t =1.969,3.122,4.324;P =0.038,0.023,0.009).After injection of 6 weeks,the epithelial basal cells (F =17.306,P =0.002) and inflammatory cells (F =34.024,P =0.000) of group A,B and C were significant changed,the differences were statistically significant.After injection of 6 weeks,the epithelial basal cells (F =3.749,P =0.042)and inflarnmatory cells(F=8.806,P =0.005) of group D,E and F were significant changed,the differences were also statistically significant.Conclusion Lacrimal duct embolization with freeze-dried human fibrin glue is effective for the xerophthalmia.

OBJECTIVE:To develop a method for simultaneous determination of 6 alkaloids in Coptis teeta.METHODS:The determination was performed on XtimateTM C18 with mobile phase consisted of 30 mmol/L ammonium bicarbonate [containing 0.1％ triethylamine and 0.7％ ammonia-acetonitrile (gradient elution)] at the flow rate of 1.0 mL/min.The detection wavelength was set at 270 nm,the column temperature was 30 ℃,and the sample size was 10 μL.RESULTS:The linear ranges of jateorrhizine,columbamine,epiberberine,coptisine,palmatine hydrochloride and berberine hydrochloride were 0.85-16.96 mg/L (r=0.999 9),1.25-24.96 mg/L(r=0.999 8),2.05-40.96 mg/L(r=0.999 9),3.65-72.96 mg/L(r=0.999 9),2.88-57.60 mg/L(r=0.999 9) and 13.25-264.96 mg/L(r=0.999 9),respectively.RSDs of precision,stability and reproducibility tests were all lower than 3.0％.Recoveries were 97.14％-102.14％ (RSD=1.93％,n=6),97.00％-102.00％ (RSD=2.06％,n=6),98.18％-101.82 ％ (RSD=1.79％,n=6),96.15％-101.28％ (RSD=2.06％,n=6),96.88％-101.88％ (RSD=1.87％,n=6),99.31％-103.76％ (RSD=1.89％,n=6),respectively.CONCLUSIONS:The method is simple,accurate,stability and reproducible,and can be used for simultaneous determination of 6 alkaloids in Coptis teeta.

Objective The effects of candesartan,an angiotensin Ⅱ type 1 receptor blocker (ARB) were investigated on advanced glycation end-products accumulation and the receptor for AGE (RAGE) expression in type 2 diabetic KK/Ta mouse kidneys.MethodsKK/Ta mice(n=72)were random divided into three groups(n=24) and it was treated with candesartan [4 mg/(kg·d)] or vehicle from 6 or 12 to 28 weeks of age.BALB/c mice(n=24) treated with vehicle were used as controls.Body weight,blood pressure,blood glucose,urinary microalbumin,urinary creatinine and serum creatinine were measured every four weeks.At 28 weeks,renal expressions of carboxymethyllysine and RAGE were evaluated by immunohistochemistry and/or competitive RT-PCR.Results KK/Ta mice developed high body weight,high blood glucose,and high urinary microalbumin/creatinine ratio in KK/Ta mice at 28 weeks of age,and it was significantly higher than that of BALB/c mice [(427.49±89.37)mg/g vs (9.54±3.25)mg/g,P＜0.01 ].Protein and mRNA expressions of RAGE were upregulated in KK/Ta kidneys with increased immunostaining intensities of carboxymethyllysine.Candesartan treatment has markedly reduced urinary microalbumin/creatinine ratio [Early treatment group (32.18±9.41)mg/g,Late treatment group (53.20±7.26)mg/g,P＜0.01 ].Treatment with candesartan down-regulated the protein and mRNA expressions of RAGE and reduced the accumulation of carboxymethyllysine.There were no significant differences between the two treatment groups (from 6 or 12 weeks).ConclusionsThe results suggest that candesartan,an ARB,reduces advanced glycation end-products accumulation and subsequent albuminuria by down-regulating RAGE expression in type 2 diabetic KK/Ta mouse kidneys.

Objective To investigate the effects of angiotensin receptor blocker (ARB)losartan on the glomerular protein expression profile of spontaneous type 2 diabetic KKAy mice by two-dimensional differential gel eleetrophoresis and MALDI-TOF mass spectrometry.Methods 8-week-old spontaneous type 2 diabetic KKAy mice were randomly divided into losartan (10 mg·kg-1·d-1 given in drinking water) treatment group and non-treatment group.Eight-week-old C57BL/6 mice were used as normal control.The glomeruli were separated by magnetic bead perfusion through thoracic aorta at age of 20 weeks,then glomerular protein was extracted.The glomerular protein expression profile was investigated by CyDyes minimal fluorescence labelling,two-dimensional differential gel electrophoresis and MALDI-TOF mass spectrometry.Results KKAy mice developed higher body weight and blood glucose,higher urinary microalbumin creatinine ratio at age of 20 weeks than C57BL/6 mice at the same age (all P＜0.05).Losartan treatment markedly reduced urinary microalbumin creatinine ratio [(539.71 ±100.23)mg/g vs (728±177.19) mg/g],attenuated mesangial expansion and the thickening of glomerular basement membrane,but had no effect on the blood glucose.By DeCyder 2-D differential analysis software,62 protein spots of differential expression were found in glomeruli between losartan treatment and non-treatment KKAy mice at age of 20 weeks.Among them,41 proteins were identified by peptide mass fingerprinting.The expressions of 28 proteins were up-regulated by losartan treatment,including glycerokinase,sulfite oxidase,glycine amidinotransferase,adenosylhomocysteinase,etc.The expressions of 13proteins were down-regulaled by losartan treatment,including 3-mercaptopyruvate sulfurtransferase,ATP synthase subunit d,60 000 heat shock protein,stress-70 protein (alternative name 75 000glucose-regulated protein,GRP75),etc.Six differcntially expressed proteins were found in glomeruli between non-treatment KKAy mice and C57BL/6 mice,and the differential expressions were suppressed by losartan treatment,including dihydrolipoyllysine-residue acetyltransferase component of pyruvate dehydrogenase complex,succinyl-CoA ligase (GDP-forming) subunit beta,mitochondrial,ATP synthase subunit d,GRP75,nucleoside diphosphate-linked moiety X motif 19 and seleniumbinding protein 1.Conclusions Losartan significantly reduces the urinary protein excretion rate and renal pathological lesion of spontaneous type 2 diabetic KKAy mice,and suppresses the differential expression of mitochondrial ATP synthase subunit d,GRP75,selenium-binding protein 1,etc in glomeruli.Losartan may play a renoproteetive role by reducing glomerular mitochondrial reactive oxygen species genesis and inhibiting oxidative stress.

Objective To examine the expressions of FHIT gene and HER_2 in non-small cell lung cancer(NSCLC)and the relationship between the expressions and the clinicopathological features of NSCLC, and to investigate the relationship between FHIT protein and HER_2 expression. Methods FHIT protein and HER_2 expressions in 50 lung cancer cases and 21 adjacent non-cancer tissues were performed by immunohistochemistry. And the positive rates of FHIT and HER_2 were measured. Results The positive rates of FHIT protein were 30.00% in lung cancer tissues. The expression levels of FHIT were significantly lower in lung cancer tissues than that in non-cancer lung tissues (P

We report the prenatal diagnosis and treatment of a case of primary right pulmonary deficiency. A routine ultrasound examination at 23 +6 weeks of gestation found an absent right lung, enlarged left lung, and dextroposition of the heart in a female fetus. Karyotype and chromosome microarray analysis of the amniotic fluid was normal. After multidisciplinary consultation, the pregnant woman chose to continue the pregnancy and had a normal delivery at 39 +2 gestational weeks. Apgar scores were ten at both 1 min and 5 min after birth, with no abnormal appearance of the baby, or signs of thoracic collapse other than no obvious respiratory sounds in the right lung. Both postnatal chest X-ray and CT scan indicated an undeveloped right lung. Cardiac ultrasonography revealed the absence of the right pulmonary artery and vein. The patient's breathing was stable during hospitalization and she was discharged eight days after birth. The infant was followed up at five months, and the growth and development were comparable to children of the same age.