Application of artificial intelligence for community-based diabetic retinopathy detection and referral / 中华实验眼科杂志

Objective:To evaluate the value of applying an artificial intelligence (AI) system for diabetic retinopathy (DR) detection and referral in community.Methods:A diagnostic test study was conducted.Four hundred and twenty-one patients (812 eyes) diagnosed with diabetes in three Dongguan community healthcare centers from January 1, 2020 to December 31, 2021 were enrolled.There were 267 males, accounting for 63.42% and 154 females, accounting for 36.58%.The subjects were 18-82 years old, with an average age of (51.72±11.28) years.The disease course of the subjects was 0-30 years, with an average course of 3.00 (1.00, 7.00) years.At least one macula-centered 50-degree fundus image was taken for each eye to build a DR image database.All the images were independently analyzed by an AI-assisted diagnostic system for DR, trained and qualified community physicians and ophthalmologists to make diagnosis including with or without DR, referable diabetic retinopathy (RDR) and referral recommendation or not.With diagnoses from ophthalmologists as the standard, sensitivity and specificity of the AI system in detecting DR and RDR were evaluated.The consistency and effective referral rate of the AI system and community physicians in detecting DR, especially in detecting RDR were evaluzted.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of Dongguan Tungwah Hospital (No.2019DHLL046).Results:Of 812 eyes, 242 eyes were diagnosed with DR, including 23 with mild nonproliferative diabetic retinopathy (NPDR), 120 with moderate NPDR, 60 with severe NPDR and 39 with proliferative diabetic retinopathy (PDR). The other 570 eyes were diagnosed without DR.The sensitivity/specificity of AI system to detect DR and RDR was 87.60%/97.89% and 90.41%/96.29%, respectively.Compared with the ophthalmologists' diagnosis, the Cohen' s Kappa statistic of AI system to detect DR/RDR was 0.87/0.87, which was lower than 0.93/0.98 of community physicians.Among the referral-recommended cases by ophthalmologists, the effective referral rate of the AI system was 90.87% (199/219), which was higher than 89.50% (196/219) of community physicians, without statistically significant difference ( P=1.000). Conclusions:The AI system shows high sensitivity, specificity and consistency in DR detection, especially in RDR.The AI system is better in recognizing RDR than trained community physicians.

Relationship between the offset of a laser-assisted flap using the WaveLight FS200 femtosecond laser and the clinical results / 中华实验眼科杂志

Objective@#To evaluate the relationship between the offset of a laser-assisted flap using the WaveLight FS200 femtosecond laser and the clinical results after femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK).@*Methods@#In this prospective cohort study, 125 patients who underwent FS-LASIK for myopia by WaveLight FS200 femtosecond laser from June 2017 to July 2018 at the Tungwah Ophthalmic Center were divided into two groups according to the offset of the corneal flap from the pupil center: the no-offset group (57 eyes) and the certain-offset group (68 eyes); the baseline data, including age, sex, uncorrected visual acuity(UCVA), spherical degree, and central corneal thickness were matched in the two groups.UCVA, residual astigmatism, spherical degree, corneal curvature and aberration were observed 1 week and 1 month after surgery.The study protocol was approved by the Ethics Committee of Tungwah Hospital of Sun Yat-Sen University (No.2017DHLL004). Written informed consent was obtained from each subject prior to entering the study cohort.@*Results@#UCVA, corneal curvature, spherical degree, spherical and corneal aberration between the two groups were not significantly different (all at P>0.05). One week after surgery, the residual astigmatism was ≤-0.50 D in 50 eyes (87.7%, 50/57)and >-0.50 D in 7 eyes (12.3%, 7/57) in the no-offset group; the residual astigmatism was ≤-0.50 D in 50 eyes (73.5%, 50/68) and >-0.50 D in 18 eyes (26.5%, 18/68) in the no-offset group.The residual astigmatism between the two groups 1 week after surgery was significantly different (χ2=3.902, P=0.048), and there was no significant difference in residual astigmatism 1 month after surgery (χ2=2.031, P=0.068). The trefoil of the no-offset group was statistically less than of the certain-offset group at 1 week and 1 month after surgery (0.05[0.04, 0.08]vs.0.06[0.04, 0.10]; 0.05[0.03, 0.07]vs.0.06[0.05, 0.09])(Z=-2.245, P=0.022; Z=-2.370, P=0.018). The spherical aberration and coma were not significantly different between the two groups at 1 week and 1 month after surgery (both at P>0.05).@*Conclusions@#The offset from femtosecond laser-assisted flap by WaveLight FS200 has no effect on long-term visual acuity or residual astigmatism.Some patients may have different visual experiences because of the residual astigmatism and higher order aberration during the early postoperative stage.

Relationship between the offset of a laser-assisted flap using the WaveLight FS200 femtosecond laser and the clinical results / 中华实验眼科杂志

Objective To evaluate the relationship between the offset of a laser-assisted flap using the WaveLight FS200 femtosecond laser and the clinical results after femtosecond laser-assisted laser in situ keratomileusis (FS-LASIK).Methods In this prospective cohort study,125 patients who underwent FS-LASIK for myopia by WaveLight FS200 femtosecond laser from June 2017 to July 2018 at the Tungwah Ophthalmic Center were divided into two groups according to the offset of the corneal flap from the pupil center:the no-offset group (57 eyes) and the certain-offset group (68 eyes);the baseline data,including age,sex,uncorrected visual acuity (UCVA),spherical degree,and central corneal thickness were matched in the two groups.UCVA,residual astigmatism,spherical degree,corneal curvature and aberration were observed 1 week and 1 month after surgery.The study protocol was approved by the Ethics Committee of Tungwah Hospital of Sun Yat-Sen University (No.2017DHLL004).Written informed consent was obtained from each subject prior to entering the study cohort.Results UCVA,corneal curvature,spherical degree,spherical and corneal aberration between the two groups were not significantly different (all at P＞0.05).One week after surgery,the residual astigmatism was ≤-0.50 D in 50 eyes (87.7％,50/57) and ＞ -0.50 D in 7 eyes (12.3％,7/57) in the no-offset group;the residual astigmatism was ≤-0.50 D in 50 eyes (73.5％,50/68) and ＞-0.50 D in 18 eyes (26.5％,18/68) in the no-offset group.The residual astigmatism between the two groups 1 week after surgery was significantly different (x2 =3.902,P =0.048),and there was no significant difference in residual astigmatism 1 month after surgery (x2 =2.031,P =0.068).The trefoil of the no-offset group was statistically less than of the certain-offset group at 1 week and 1 month after surgery (0.05 [0.04,0.08] vs.0.06[0.04,0.10];0.05[0.03,0.07] vs.0.06[0.05,0.09]) (Z=-2.245,P--0.022;Z=-2.370,P=0.018).The spherical aberration and coma were not significantly different between the two groups at 1 week and 1 month after surgery (both at P＞0.05).Conclusions The offset from femtosecond laser-assisted flap by WaveLight FS200 has no effect on long-term visual acuity or residual astigmatism.Some patients may have different visual experiences because of the residual astigmatism and higher order aberration during the early postoperative stage.

Induction of Rhesus Keratinocytes into Functional Ameloblasts by Mouse Embryonic Dental Mesenchyme

Fast progresses in stem cell-based tooth tissue engineering have been achieved in recent years in several animal models including the mouse, rat, dog, and pig. Moreover, various postnatal mesenchymal stem cells of dental origin have been isolated and shown capable of differentiating into odontoblasts and generating dentin. Meanwhile, human keratinocyte stem/progenitor cells, gingival epithelial cells, and even iPSC-derived epithelium have been demonstrated to be able to differentiate into functional ameloblasts. Translational medicine studies in the nonhuman primate are irreplaceable steps towards clinical application of stem cell-based tissue engineering therapy. In the present study, we first examined the epithelial stem cell markers in the rhesus skin using immunostaining. Keratinocyte stem cells were then isolated from rhesus epidermis, cultured in vitro, and characterized by epithelial stem cell markers. Epithelial sheets of these cultured keratinocytes, which were recombined with E13.5 mouse dental mesenchyme that possesses odontogenic potential in the presence of exogenous FGF8, were induced to differentiate into enamel-secreting ameloblasts. Our results demonstrate that in the presence of appropriate odontogenic signals, rhesus keratinocytes can be induced to gain odontogenic competence and are capable of participating in odontogenesis, indicating that rhesus keratinocytes are an ideal epithelial cell source for further translational medicine study of tooth tissue engineering in nonhuman primates.

Evaluating diabetic macular edema quantificationally by optical coherence tomography / 眼科新进展

Objective To observe the changes of four quantitative indexes of diabetic macular edema by using optical coherence tomography.Methods Eighty-nine patients (155 eyes) with diabetic retinopathy were included in this project and were divided into two groups according to the present of diabetic macular edema:Negative group (56 cases,100 eyes) and positive group(33 cases,55 eyes).In addition,23 cases (42 eyes) of normal volunteers constituted the normal control group.All the objects accepted an optical coherence tomography examination and the indexes including central retinal thickness (CRT),subfoveal choroidal thickness (SFCT) and integrity of external limiting membrane(ELM) as well as inner/outer segment (IS/OS) were measured and analyzed.Results The average CRT of positive group (219.048 ± 16.798) μm was significantly thicker than that of control group(217.775 ± 26.866) μm and negative group (280.418 ±74.187) μm (P ＜0.001).Mean SFCT among control group (312.893 ±140.559) μm,negative group (302.080 ± 125.287) μm and positive group (293.745 ±140.517) μm had no statistical significance (P =0.781).There were 3 eyes with disrupted ELM layer in the negative group and 8 eyes in the positive group.Difference between them was proved to be significant (P =0.019).Similarly,the integrity of IS/OS layer had significant difference between negative group (5 eyes disrupted) and positive group (19 eyes disrupted) (P ＜ 0.001).Conelusion CRT of patients with diabetic macular edema is always increased and the integrity of ELM or (and) IS/OS can be disrupted in many cases.Indexes including CRT and the integrity of ELM or (and) IS/OS can be used to evaluate the severity of diabetic macular edema quantificationally.

Rosiglitazone down-regulates lipopolysaccharide-induced expression of CD40 and intercellular adhesion molecule 1 in rat peritoneal mesothelial cells through a NF-κB dependent mechanism / 中华肾脏病杂志

Objective To investigate the effect and mechanism by which PPARγ ligand, rosiglitasone, regulates the expression of CD40 and intercellular adhesion molecule 1 (ICAM-1) in the rat peritoneal mesothelial cells (RPMCs) induced by lipopolysaccharide (LPS). Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were randomly divided into groups as follows: medium, LPS (5 mg/L), LPS (5 mg/L)+BAY11-7085(5 μmol/L, NF-κB inhibitor), rosiglitazone (10 μmol/L or 20 μmol/L, peroxisome proliferator-activated receptor γ activator), LPS (5 mg/L)+rosiglitazone (10 μmol/L)+GW9662 (3 μmol/L, peroxisome proliferator-aetivatcd receptor γ antagonist), and LPS (5 mg/L)+vehicle (DMSO 0.2 ml/L). The expressions of CD40 and ICAM-1 RNA in RPMCs were examined by RT-PCR after 3 hour treatment, and the protein expressions of CD40, ICAM-1, p-NF-κB p65 and p-IκBα were examined by Western blot or immunofluorescence after 24 hour treatment. Results Following treatment with LPS, both the expressions of CD40 and ICAM-1 protein in RPMCs were up-regulated significantly (P<0.05), and the phosphoralation of p65 was increased greatly (1.10±0.17 vs 0.55±0.06, P<0.05). BAY11-7085 (5 μmol/L) significantly decreased the protein expression of p-p65 (0.22±0.11 vs 1.10±0.17, P<0.01), CD40 (0.34±0.02 vs 0.50±0.06, P<0.05) and ICAM-1 (0.35±0.16 vs 0.74±0.03, P<0.05). Pretreated with rosiglitazone for 3 h then added with LPS for 1 h, the levels of p-p65, CD40 and ICAM-1 in RPMCs were significantly decreased compared with those of LPS group (0.77±0.08 vs 0.90±0.10, P<O.01; 0.79±0.16 vs 0.99±0.06, P<0.05; 0.83±0.20 vs 1.22±0.13, P<0.05). However, for the group pretreated with rosiglitazone(10 μmol/L) and GW9662(3 μmol/L) for 3 h then added with LPS, the levels of p-p65 in RPMCs did not change significantly compared with those of rosiglitazone-pretreated group. The expressions of CD40 and ICAM-1 in RPMCs were significantly increased compared with those of rosiglitazone-pretreated group (0.95±0.19 vs 0.79±0.16, and 1.04±0.24 vs 0.83±0.20, P<0.05). Conclusion Rosiglitazone can decrease LPS-induced expression of CD40 and ICAM-1 in RPMCs by inhibition of NF-κB activation, which suggests that rosiglitazone may mediate its antiinflammatory effect through a NF-κB dependent mechanism.

Correlation of peritoneal albumin leakage with malnutrition-inflammation-atherosclerosis syndrome in continuous ambulatory peritoneal dialysis patients / 中华肾脏病杂志

Objective To investigate the impact of peritoneal albumin leakage on malnutrition-inflammation-atherosclerosis (MIA) syndrome in continuous ambulatory peritoneal dialysis (CAPD) patients. Methods A cross-sectional study of a cohort of 130 CAPD patients without edema or active infection was performed. In order to identify peritoneal transport characteristics in CAPD patients, a standard peritoneal equilibration test (PET) was carried out. For malnutrition and inflammation, serum albumin and high-sensitivity C-reactive protein (hs-CRP) levels were measured. Mean-carotid artery intima media thickness (IMT) was used to determine atherosclerosis. Residual glomerular filtration rate (rGFR) was defined as the average of 24-hour urinary urea and creatinine clearances. Results Pearson and Spearman correlation analysis showed that peritoneal albumin leakage amount was positively correlated with age, body mass index, night dwell time, blood glucose, 4 h D/P creatinine levels and hs-CRP levels (r=0.204, P<0.05 ;r=0.314, P<0.01; r=0.265, P<0.01; r=0.212, P<0.05; r=0.401, P<0.01; r=0.216, P<0.05); whereas it was negatively correlated with diastolic perssure, serum albumin levels, glucose level of dialyzate and peritoneal Kt/V (r=-0.209, P<0.05; r=-0.123, P<0.05; r=-0.271, P<0.01; r=-0.212, P<0.01). Overall, there was no correlation between peritoneal albumin leakage and IMT. Patients was significantly greater (P<0.01), and there was a positive correlation between peritoneal albumin leakage amount and IMT (r=0.650, P<0.01). Conclusions Peritoneal albumin leakage is significantly associated with peritoneal transport characteristics, malnutrition and inflammatory state in CAPD patients. High peritoneal albumin leakage amount is a risk factor for atherosclerosis in patients with rGFR less than 1 ml·min-1(1.73 m2)-1.

Role of rat organic anion transporter 1 in renal cellular uptake of aristolochic acid Ⅰ and the induction of cellular toxicity / 中华肾脏病杂志

Objective To investigate the role of rat organic anion transporter 1 (OAT1, SLC22A6) in the renal cellular uptake of AA Ⅰ and its impact on cellular toxicity. Methods HEK-293 cells were transfeeted with rat OAT1 cDNA or empty vectors. The over-expression of rOAT1 was confirmed by Western blot analysis and its activity was validated by using para-aminohippurate (PAH) as a probe. Cellular apoptosis was examined by flow cytometery using propodium iodode (PI) and annexin V-FITC staining. Results Concentration-and time-dependent intracellular accumulation of AA Ⅰ was observed in rOATl-transfected HEK-293 cells. After treatment with AA Ⅰ at the concentrations of 40 mg/L, 80 mg/L, 120 mg/L and 160 mg/L,respectively, for 45 min, the intracellular concentrations of AA Ⅰ in rOAT1-transfected HEK-293 cells were higher than those in controls (P<0.05). After treatment with AA Ⅰ (120 mg/L for 30 min, 60 min, 90 min and 120 min, respectively, the intracellular concentrations of AA Ⅰ in rOAT1-transfected HEK-293 cells were higher than those in controls (P<0.05). PAH significantly reduced the intracellular accumulations of AA Ⅰ in rOAT1-transfected HEK-293 cells. After treatment with AA Ⅰ at the concentrations of 40 mg/L, 80 mg/L, 120 mg/L and 160 mg/L respectively for 35 min, the intracellular accumulations of AA Ⅰ in rOAT1-transfected HEK-293 cells that treated with PAH were lower than those that were not treated by PAH. Cellular apoptosis and caspase-3 expression in rOAT1-transfeeted HEK-293 cells were significantly up-regnlated as compared to controls (P<0.05). Conclusion rOAT1 is involved in the cellular uptake of AA Ⅰ which leads to increased epithelial apoptosis. Further studies are suggested to investigate the role of human OAT in the disposition of AA and its toxicological consequences.

Expression of PPAR-γ,TLR4 and activation of STAT1 signal protein in rats with peritoneal dialysis related acute peritonitis induced by LIPS / 中华微生物学和免疫学杂志

ated acute peritonitis induced by LPS in rats.

Up-regulation of CD40 and ICAM-1 expression treated with lipopolysaccharide through the activation of NF-κB in rat peritoneal mesothelial cells / 中华微生物学和免疫学杂志

Objective To investigate the expression of CD40 and intercellular adhesion molecule-1 (ICAM-1) treated with lipopolysaccharide (LPS) in rat peritoneal mesothelial cells(RPMC) and the role of NF-κB signal transduction pathway. Methods RPMCs were harvested from Sprague-Dawley rat peritoneal cavity and maintained under defined in vitro conditions. The cells were exposed respectively to different concentrations of LPS for 12 h or treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of CD40 and ICAM-1, the RPMCs were treated with LPS (5 μg/ml) for different time points. To observe the effect of LPS on the expression of NF-κB and p-NF-κB protein, the RPMCs were treated by LPS or pretreated with BAY11-7085 (5 μmol/L or 1 μmol/L ) for 3 h, then treated with LPS for another 3 h, respectively. Expression of CD40 and ICAM-1 mRNA was examined by RT-PCR. Expression of NF-κB and p-NF-κB protein was detected by Western blot. Results Compared with medium control group, stimulation of RPMCs with 1 μg/ml and 5 μg/ml of LPS resulted in a significant increase in the expression of CD40 and ICAM-1 mRNA(P<0.05). 10 μg/ml of LPS had strongest effect on CD40 and ICAM-1 expression compared with that of 1 μg/ml and 5 μg/ml of LPS. Treatment with 5 μg/ml of LPS resulted in time-dependent increase in the gene level of CD40 and ICAM-1, with the peak at 3 h. However, after that time point, the gene level of them was gradually attenuated. Following treatment with LPS (5 μg/ml), the level of p-NF-κB began to increase at 15 min, gradually reached the peak at 1 h, and then decreased. But the level of p-NF-κB at 2 h was still significantly higher than that of medium control. 5 μmol/L of BAY11-7085 decreased significantly the up-regulation of CD40 and ICAM-1 induced by LPS. Conclusion LPS enhanced the expression of CD40 and ICAM-1 on RPMCs in a concentration-dependent and a time-dependent manner. LPS induced expression of CD40 and ICAM-1 depend on the NF-κB signal transduction pathway.

Effects of PPARγ, agonist on the expression of PPARγ. toll-like receptor 4 and STAT1 signal protein activation in rats with peritoneal dialysis-related acute peritonitis / 中华肾脏病杂志

Objective To explore the effects of peroxisome proliferator-activated receptorγ (PPARγ) agonist rosiglitazone and 15-deoxy-delta-12,14-prostaglandin J2 (15d-PGJ2) on the expression of PPARγ, toll-like receptor 4 (TLR4) and the activation of STAT1 as well as the local inflammation reaction of abdominal cavity in sprague dawley (SD) rats with peritoneal dialysis- related acute peritonitis induced by lipopolysaccharide (LPS). Methods Twenty-four male SD rats were equally randomized to four groups(n=6 each): control group, injected with 4.25% dextrose peritoneal dialysate (PDF) via abdominal cavity(90 ml/kg); LPS group, injected with LPS(1 mg/kg) via abdominal cavity 4 hours later follewed by PDF injection; rosiglitazone plus LPS group (Rosi group), preconditioned with rosiglitazone (20 mg·kg-1·d-1) by intragastric way for 3 days, then injected with LPS and PDF via abdominal cavity; 15d-PGJ2 plus LPS group (15d-PGJ2 group), preconditioned with 15d-PGJ2 (0.3 mg·kg-1·d-1)via abdominal cavity injection for 3 days, then injected with LPS and PDF via abdominal cavity. The rats were killed 4 hours after PDF injection, IL-6 level in abdominal dropsy was determined by ELISA. Peritoneum tissue was stained by Masson. Leucocyte count in abdominal dropsy was performed. The mRNA expression of PPARγ and TLR4 in peritoneum tissue was determined by RT-PCR; the protein expression of PPARγ, TLR4, p-STAT1 and STAT1 in peritoneum tissue was analyzed by Western blot. Results IL-6 level of abdominal dropsy in LPS group [median 268.53 (range 201.87-335.19) ng/L] was significantly higher than that of control group [median 147.62 (range 130.60-164.64) ng/L] (P<0.01). The IL-6 level of abdominal dropsy in Rosi group [median 110.20 (range 77.60-142.80) ng/L] was significantly lower than that of LPS group (P<0.05). Compared to that of control group, the edematous degree of peritoneum in LPS group was significantly severer, meanwhile, mRNA and proteins expression of PPARγ and TLR4 in rat peritoneum were also significantly higher (P<0.05, P<0.01). Compared to that of LPS group, the edematous degree of peritoneum in Rosi group was lighter, the expression of PPARγ and TLR4 mRNA was significantly up-regulated (P<0.05), meanwhile their proteins expression was down-regulated (P<0.05); and in 15d-PGJ2 group, the edematous degree of peritoneum, the expression of PPARγ mRNA and protein was also decreased (P<0.05), but TLR4 mRNA expression was up-regulated (P<0.01), however, its protein expression was down-regulated (P<0.05). There were no significant differences in leucocyte count of abdominal dropsy among the four groups. The p-STAT1 expression in the rats peritoneum induced by LPS was markedly increased by both rosiglitazone and 15d-PGJ2 (P<0.01). Conclusions Both rosiglitazone and 15d-PGJ2 can down-regnlate the inflammatory reaction in rat peritonitis induced by LIPS, which may be involved in modulating the expression of associated functional protein during LPS signal pathway.

JNK regulates epithelial mesenchymal transition induced by transforming growth factor β1 in rat peritoneal mesothelial cells / 中华肾脏病杂志

Objective To investigate the role of C-Jun N-terminal kinase (JNK) in epithelial mesenchymal transition (EMT) induced by transforming growth factor β1 (TGF-β1) in rat peritoneal mesothelial cells(RPMCs). Methods RPMCs were harvested from the peritoneum of male Sprague-Dawley rats, then cultured in DMEM/F12 medium with 15% (V/V) FBS. After stimulation with TGF-β1, the expression of a-smooth muscle actin (α-SMA), E-cadherin and collagen I were detected in RPMCs. In some groups, the ceils were pretreated with SP600125, a specific inhibitor of JNK, for 4 hours before incubation with TGF-β1. The protein expression of phosphorylated JNK was detected by Western blotting. The mRNA and protein expression ofα-SMA, E-cadherin and collagen I were examined with RT-PCR and Western blotting, respectively.The intracellular distribution and expression of α-SMA was determined by indirect immunofluorescence. Results TGF-β1 could significantly increase the expression of α-SMA and collagen I, and decrease the expression of E-cadherin in RPMCs. TGF-α1 could stimulate the expression of phosphorylated JNK at 5 minutes with the peak at 10 minutes (P<0.01). The addition of SP600125 effectively inhibited TGF-β1-induced high expression of α-SMA and collagen I (P<0.05), and prevented TGF-β1-induced down-regulation of E-cadherin expression in RPMCs (P<0.05). The indirect immunofluorescence showed that the expression of intracellular α-SMA in RPMCs stimulated by TGF-β1 for 48 h increased significantly, which could be inhibited by SP600125. Conclusions JNK regulates epithelial mesenchymal transition induced by TGF-β1 in rat peritoneal mesothelial cells. JNK inhibitor may be used as a novel therapeutic agent for peritoneal fibrosis.

Expression of regulatory T cells and helper T cells in human IgA nephropathy and its significance / 中华肾脏病杂志

Objective To investigate the effects of CD4+CD25high regulatory T cells(Treg)and the imbalance of helper T lymphocyte subsets(Th1/Th2)on the immunological mechanism of IgA nephropathy(IgAN)patients. Methods The percentage of Treg and helper T cells subpopulation (Th1/Th2)in the peripheral blood of IgAN patients and healthy controls was examined by flow cytometry.The FOXP3 expression was detected through intracellular staining.The correlation of Treg or Th1/Th2 with clinical parameters of IgAN was analyzed by Spearman or Pearson rank correlation test. Results The percentages of Treg and Th2 cells were significantly higher in peripheral blood of IgAN patients compared to that of healthy controls[Treg (2.14±0.82)%vs[1.59±0.53)%,Th2(2.57±0.72)%vs(1.81±1.10)%,all P＜0.05].Th1/Th2 ratio was significantly reduced in IgAN patients(5.75±1.89 vs 12.73±9.79,P＜0.05).The percentage of circulating Treg cells was positively correlated with serunl IgA concentration(r=0.397,P＜0.05),and was negatively correlated with eGFR(r=-0.376,P＜0.05).The percentage of circulating Th2 cells was positively correlated with serum IgA(r=0.468,P＜0.05). Conclusions There is a disorder of T lymphocyte population in the peripheral blood of IgAN patients.The increased Treg and Th2 cells may play an important role in the pathogenesis of IgAN.

Role of TLR4 expression induced by angiotensin Ⅱ in LPS-induced CD40 expression in rat peritoneal mesothelial cells / 中华肾脏病杂志

Objective To investigate the effects of angiotensin Ⅱ (Ang Ⅱ) on the expression of TLR4 and its role in lipopolysaccharide (LPS)-induced NF-κB activation and CD40 expression in rat peritoneal mesothelial cells (RPMCs). Methods RPMCs were harvested from Spragne-Dawley rat peritoneal cavity and maintained under defined in vitro condition. The cells were treated with Ang Ⅱ at different concentrations (10-9, 10-8, 10-7, 10-6 mol/L) and exposed to Ang Ⅱ (10-7 mol/L) for different times (1, 2, 4, 8, 12, 24, 48 h for mRNA and 6, 12, 24, 36, 48 h for protein, respectively). Meanwhile, the influence of AT1 receptor antagonist (AT1R, losartan, 10-5 mol/L) and AT2 receptor blocker (AT2R, PD123177, 10-5 mol/L) on the TLR4 induced by Ang Ⅱ was observed. After synchronization for 24 hours, the cells were randomly assigned to four groups: the control group, the Ang Ⅱ (10-7 tool/L) group, the LPS (1 mg/L) group, the Ang Ⅱ (10-7 mol/L) plus LPS (1 mg/L) group, which were used to investigate the effects of Ang Ⅱ on the NF-κB activation and CD40 expression induced by LPS. The mRNA expression of TLR4 and CD40 was measured by RT-PCR and the protein abundance of TLR4, NF-κB p65, phospho-p65, IKBα and phospho-IκBα were analyzed by Western blot. Immunofluorescence was performed to determine the subcellular localization of p65 subunit of NF-κB. Results (1) Treatment of RPMCs with Ang Ⅱ resulted in a concentration-dependent increase in the expression of TLR4. Ang Ⅱ at 10-9, 10-8, 10-7 and 10-6 mol/L increased TLR4 mRNA expression by 70.5%, 89.5%, 102.9%, and 121.9%, respectively and protein expression by 12.1%, 27.7%, 51.2%, and 41.6%, respectively (P<0.01). Treatment of RPMCs with 10-7 mol/L Ang Ⅱ resulted in a time-dependent increase in the expression of TLR4, with the peak of mRNA expression at 8 and 12 h (P<0.01) and the protein expression at 12 and 24 h (P<0.01). (2) Losartan antagonized Ang Ⅱ-stimulated expression of TLR4 by 33.5% (P<0.05), PD123177 had no such effect (P0.05). (3) Treatment of RPMCs with LPS (1 mg/L) for 60 rain significantly increased the ratio of phospho-IκBα to IκBα by 362.6% (P< 0.01) , phospho-p65 to p65 by 67.4% (P<0.05), and LPS (1 mg/L) for 4 h significantly increased the expression of CD40 mRNA by 299.9% (P<0.01) compared to the control group. In comparison to the LPS (1 mg/L) group, preincubation of RPMCs with AngⅡ (10-7 mol/L) for 24 h then treated with LPS (1 mg/L) for 60 rain significantly increased the ratio of phospho-IκBα to IκBα by 49.1% (P<0.01), phospho-p65 to p65 by 29.3%(P<0.05), and LPS (1 mg/L) for 4 h significantly increased the expression of CD40 mRNA by 56.8%(P<0.01). (4) The p65 subunit of NF-κB was dominantly distributed in the cytoplasm in the control and Ang Ⅱ group. Following exposure to LPS for 60 min, p65 subunit labeling was upregulated and translocated into the nuclei. A significantly increased nuclear staining of p65 in ceils treated with Ang Ⅱ plus LPS were observed. Conclusions Ang Ⅱ induces the expression of TLR4 in dose- and time-dependent manner in RPMCs, resulting in enhanced NF-κB signaling and induction of CD40 expression, Locally produced Ang Ⅱ in the peritoneum may play an amplified role in LPS-induced peritoneal inflammation.

Major problems in Changzhou City's community health services and countermeasures / 中华医院管理杂志

An investigation has been conducted on the pilot projects of community health services in Chang zhou City and an analysis made of several issues hindering the full functioning of the services. The following sugges tions are then put forward. ① Leadership in community health services should be strengthened in real earnest. ② Methods of compensating funds for commmunity health services should be further improved. ③An effective management system and operating mechanism should be established and perfected. ④The building of a contingent of community health services personnel should be enhanced. ⑤Educational campaigns should be conducted at various levels. ⑥A good job should be done of community health services along the lines of reform.

Glutathione depletion inhibits angiotensinⅡ-induced activation of c-Jun/ATF-2 and NF-?B in cultured macrophages / 中国病理生理杂志

AIM: The aim of this study was to reveal the regulatory role of glutathione (GSH) in the transcriptional activity of activating transcription factor-2 (c-Jun/ATF-2) and nuclear factor-?B (NF-?B) of macrophages induced by angiotensin Ⅱ(AngⅡ). METHODS: Macrophage intracellular GSH was determined by fluorophotometry, and buthionine-[S,R]-sulfoximine(BSO)was used for depletion of intracellular GSH. The phosphorylation of c-Jun/ATF-2 and expression of NF-?B p65 were determined by immunoblot, and the activity of NF-?B was determined by electrophoresis mobility shift assay (EMSA). The c-Jun/ATF-2 was also determined by Immunohistochemical staining. RESULTS: The GSH content in the macrophage was decreased in cells that were lipid-peroxidized with AngⅡ (1.0 (?mol/L)) for 30 min and 60 min, respectively, followed by an adaptive GSH increase in the presence of AngⅡ (1.0 (?mol/L)) for longer time. In parallel, exposure to AngⅡfor 60 min also decreased macrophage GSH content in a dose-dependent manner. The GSH of RAW 264.7 cells were depleted by BSO, a specific inhibitor of GSH synthesis, and incubation for 18 h with 0.5 mmol/L BSO was sufficient for complete depletion of intracellular GSH. The phosphorylation of c-Jun/ATF-2 could be induced by the AngⅡ (1.0 (?mol/L)), whereas it did not occur in glutathione-depleted RAW 264.7 macrophages. The activation of NF-?B could also be induced by the AngⅡ (1.0 (?mol/L)), but it did not occur in glutathione-depleted RAW 264.7 macrophages. CONCLUSION: These data provide evidences that the intracellular glutathione redox may participate in the regulation of transcription activity of c-Jun/ATF-2 and NF-?B in macrophages. [

Activation of smad signaling and collagenⅠ synthesis in NRK52E cells induced by advanced glycation end products / 中国病理生理杂志

AIM: To investigate the effects of advanced glycation end products on activation of Smad signaling pathway and collagenⅠ synthesis in proximal tubular epithelial cells.METHODS: Advanced glycation end products(AGE-BSA) were prepared by incubation of bovine serum albumin(BSA) with D-glucose.Normal rat proximal tubular epithelial(NRK52E) cells were cultured in RPMI-1640 medium with AGE-BSA.Phosphorylation and nuclear translocation of Smad2/3 were examined by immunocytochemistry.Levels of TGF-?_1 in supernatant of cell culture were measured by enzyme-linked immunosorbent assay(ELISA).Expression of TGF-?_1,Smad2,Smad3 and Smad7 mRNA were detected by RT-PCR.Expression of ?-SMA,E-cadherin and collagenⅠproteins were detected by Western blotting.RESULTS: AGE-BSA induced Smad2/3 phosphorylation and nuclear translocation,two peaks occured at 30 min(68% vs 16%,P

Association and function study of tumor necrosis factor receptorⅡ position 196 polymorphism with systemic lupus erythematosus / 中国病理生理杂志

AIM: To investigate the association of gene polymorphism at position 196 of tumor necrosis factor receptor Ⅱ (TNFRⅡ) with systemic lupus erythematosus (SLE) in Chinese, and establish recombinant retroviral vector to analyze the function of the TNFRⅡ 196M/R. METHODS: The genotype at position 196 of TNFRⅡ was determined by PCR-RFLP in 106 SLE patients and 119 healthy controls in china. Human TNFRⅡ196M cDNA were amplified by PCR and cloned into PMD18-T vector. Then, PMD18-TNFRⅡ196R was induced by site-directed mutagenesis. The recombinant T vector, PMD18-TNFRⅡ196M and PMD18-TNFRⅡ196R, were subcloned into retroviral vector PLXSN. Both normal and variant were transfected into rat mesangial cell. The effects of TNF? on production of sTNFRⅡ and IL-6 were study by ELISA. RESULTS: (1) The frequency of TNFRⅡ196R allele was significantly higher than those in controls (35.2% vs 14.3%, P

Recombinant human defensin ?1 induces proliferation of rat glomerular mesangial cells in vitro / 中国病理生理杂志

AIM:To study the effects and mechanism of recombinant human defensin ?1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS:The influences of defensin ?1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase(MEK)inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin ?1,the phosphorylation of extracellular signal regulated kinase(ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS:Defensin ?1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h(P20 mg/L decreased cell proliferation.The cell proliferation induced by defensin ?1 was inhibited by U0126.Stimulation of the cells with defensin ?1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin ?1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h(P

Expression and function of CD40 on rat peritoneal mesothelial cells / 中华肾脏病杂志

Objective To determine whether peritoneal mesothelial cells express CD40 and to investigate potential mechanism by which CD40-CD40 ligand(CD40L) interaction may be involved in the inflammation of peritoneal membrane. Methods Rat peritoneal mesothelial cells (MC) were harvested from peritoneal cavity and maintained under defined in vitro conditions. Expression of CD40 on MC under normal culture or stimulation with interferon-?(IFN-?), tumor necrosis factor-?(TNF-?), interleukin(IL)-1 was examined by RT-PCR and FACS analysis. After activation with CD40 mAb, the expression of intercellular adhesion molecule-1 (ICAM-1) on MC was analyzed by FCAS. Results MC cultured in vitro expressed CD40 constitutively. The expression of CD40mRNA and CD40 protein up-regulated markedly following the stimulations with IFN-?, but not with IL-1, TNF-?. The expression of ICAM-1 on MC was significantly increased after activation of CD40 with IFN-? and CD40mAb. Conclusions MC functionally expresses CD40. The interaction of CD40 on MC and CD40L+ cells in peritoneal cavity may play an important role in peritoneal local defense and may be involved in the inflammation process of peritoneum.