Background Allograft rejection is a main cause of failure of penetrating keratoplasty,especially in the patient with high risk of rejection condition.Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization,but these factors did not explain all the phenomena of allograft rejection.Research found that immune cells appeared in iris and ciliary body when rejection occurred,but the relationship between these immune cells and allograft rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty.Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group (60 mice) and blank control group (10 mice).Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/c mice as the recipients.The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde.The mice were sacrificed and iris and ciliary tissue were obtained 5,10 days and rejection after surgery.Immunohistochemistry and reverse transcription PCR (RT-PCR) was used respectively to detect the expression diversities of occludin,zonula occludens protein-1 (ZO-1),matrix metalloproteinase-9(MMP-9),major histocompatibility complex-Ⅱ (MHC-Ⅱ),and CCR5,CCR7 and their mRNA in iris and ciliary body.Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount,and absorbance of target genes (A values).The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research.Results The mean survival time of corneal gratts was (17±3) days after operation.The mean score was 0.6 in 5 days and 0.5 in 10 days,and 3.3 in 18 days after operation.Expression of ZO-1 reduced significantly,and that of MMP-9 increased obviously at the time of rejection.MHC Ⅱ + cells were scattered in iris and ciliary body in normal mice,and the number of the positive cells (cells/field) was increased after operation with a peak value when rejection occurred.A significant difference was seen between normal mice and rejection mice (1559.67±350.29 vs.4021.83±495.18) (P=0.000).The expressions of occludin mRNA and ZO-1 mRNA in the iris and ciliary body decreased obviously in the rejection mice.Compared with normal mice,theA value of ZO-1 and occluding were 36.74±3.13 vs.110.11±11.88 and 57.54±3.41 vs.59.90±3.50respectively,with significant differences between them (all P＜0.05).The expressions of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA in the iris and ciliary body increased gradually with the time lapse after operation and peaked when the rejection appeared.The A value of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA were significantly higher than those of normal mice (20.29±1.19 vs.2.77±0.85 for MMP-9 mRNA; 35.43±2.56 vs.9.11±0.29 for CCR5 mRNA,and 60.83±0.87 vs.0.89 ±0.95 for CCR7 mRNA) respectively (all P＜0.05).Conclusions The permeability of vascules in the iris and ciliary body increase during the allograft rejection after penetrating keratoplasty.Increased antigen presenting cells were also detected.

Objective To explore the value of color doppler ultrasonography by bed side in the early diagnosis of HIE in full term neonates.Methods The changes of cerebral parenchymal and cerebral arterial blood stream parameter on 35 cases of neonates clinically diagnosed HIE of mild and moderate degree and 40 cases of normal newborns on the 24,48 and 72 hours after birth were observed by color doppler ultrasonography by bed side.Results 1.The cerebral parenchyma was even echo in normal newborns,but it was maldistributed and reinforced in mild asphyxia neonates and it was more serious in moderate degree.The echo of cerebral parenchyma in mild degree was near normal in 48 hours after birth,while the echo of cerebral parenchyma in moderate degree was still maldistributed and reinforced in 48 and 72 hours after birth.2.There was obvious changes in the cerebral arterial blood stream parameter and hemodynamics of the asphyxia newborns compared with normals.The systolic peak velocity(Vs)and end diastolic velocity(Vd)of the cerebral arteries in mild and moderate degree were obviously lower than that of control group in 24,48 hours after birth(Pa0.05).3.Resistance index(RI)of the cerebral arteries in mild and moderate degree were higher than that of control group in 24,48 hours after birth(Pa0.05).Conclusion Color doppler ultrasonography by bed side is a convenient,noninvasive method for diagnosing HIE.

OBJECTIVETo study the antiviral constituents in the stems and leaves of Pithecellibium clypearia.

METHODThe constituents of P. clypearia were systematically separated with various chromatographic techniques in combination with antiviral activity monitoring. Their structures were elucidated by physical and chemical properties and spectral data.

RESULTSix compounds were isolated from P. clypearia and were identified as: tricetiflavan (5, 7, 3', 4', 5'-pentahydroxylflavan) (1), myricitrin (myricetin-3-O-alpha-L-rhamnopyranoside) (2), quercitrin (quercetin-3-O-alpha-L-rhamnopyranoside) (3), quereetin (4), methyl gallate (5) and gallic acid (6).

CONCLUSIONCompound 1 approximately 5 were obtained from this plant for the first time. Compound 4 was found to show an obvious anti-respiratory syncytial virus (RSV) activity.

Antiviral Agents ; chemistry ; isolation & purification ; pharmacology ; Fabaceae ; chemistry ; Flavonoids ; chemistry ; isolation & purification ; pharmacology ; Gallic Acid ; analogs & derivatives ; chemistry ; isolation & purification ; pharmacology ; Inhibitory Concentration 50 ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Plants, Medicinal ; chemistry ; Quercetin ; chemistry ; isolation & purification ; pharmacology ; Respiratory Syncytial Viruses ; drug effects

OBJECTIVETo study the effect and mechanism of Danhong injection on isolated mesenteric arterial rings in rats.

METHODAn isolated vascular ring experiment was conducted to determine the changes in tension of vascular rings with a biological signal collection and analytical system.

RESULTDanhong injection had no impact on the tension of vascular rings. Danhong injection showed a significant vasodilatation effect on treated arteria rings of norepinephrine, and no remarkable impact was made on the effect without endothium. It showed notable effect on blood vessels treated with Ca(2+) and no significant impact on those treated with caffeine. It could inhibit NE-induced intracellular calcium from releasing and external calcium from inflowing. No effects of potassium channel blockers on aorta ring tensile force were found.

CONCLUSIONDanhong injection shows significant vasodilation effect, which mainly works through vascular smooth muscle. Its vasodilation effect may be related to inhibitory receptor, voltage-dependent Ca(2+)-release and IP3 receptor-mediated Ca(2 +)-influx.

Animals ; Calcium ; metabolism ; Drugs, Chinese Herbal ; administration & dosage ; In Vitro Techniques ; Injections ; Mesenteric Arteries ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley ; Vasodilation ; drug effects

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.

Objective: To prepare monoclonal antibodies (McAb) with cardiac troponin I (cTnI) which was purified from fresh human cardiac muscle within 6 h. Methods: (1) Extraction and purification of human cTnI: cTnI was purified by high salt extraction, saltless precipitation, 65℃ treatment, ammonium sulfate fractionation and DEAE-cellulose chromatography, etc. (2) Preparation of anti human cTnI McAb: The purified cTnI was injected into the spleen of BALB/c mice. The cTnI-primed spleen cells were fused with Sp2/0 myoloma cell. The McAbs anti human cTnI were obtained by screening with indirect ELISA and 3 times clone. (3)The identification of anti cTnI McAb. Results: Five hybridoma cell lines, named 3A7，3A11，3D2，3F10 and 1H9 were developed, which could secret McAb stably. The 5 McAbs all were demonstrated to be IgG2a by double gel diffusion test. The number of hybridoma chromosomes was between 92 to 110 and the chromosomes were mainly telocentric. Five kinds of ascites had no cross-reaction to LDH，CK，CK-MB ，AST and cardiac troponin T(cTnT), and their titers were between 3.2×10-6 to 1.6×10-7. Conclusion: 3D2，3F10 and 3A7,3A11,1H9 react to different epitopes of cTnI.

OBJECTIVE: To study the expression and diagnostic value of plasma miR-145 and miR-183 in children with lupus nephritis (LN). METHODS: A total of 92 children with LN who were admitted from January 2016 to May 2019 were enrolled as the LN group, among whom 17 had type II LN, 15 had type III LN, 36 had type IV LN, 18 had type V LN, and 6 had type VI LN. Forty healthy children who underwent physical examination were enrolled as the healthy control group. According to Systemic Lupus Erythematosus Disease Activity Index (SLEDAI), the 92 children with LN were further divided into a stable LN group with 34 children (SLEDAI score <10) and an active LN group with 58 children (SLEDAI score ≥10). RT-PCR was used to measure the expression of miR-145 and miR-183 in plasma. The receiver operating characteristic (ROC) curve was used to analyze the value of plasma miR-145, miR-183, and anti-dsDNA antibody in the diagnosis of LN. Pearson correlation analysis was used to investigate the correlation of the expression levels of miR-145 and miR-183 in plasma with laboratory markers. RESULTS: The LN, active LN, and stable LN groups had significantly higher levels of anti-dsDNA antibody, C-reactive protein, serum creatinine (Scr), and blood urea nitrogen (BUN) than the control group (P<0.05). The active LN group had significantly higher SLEDAI score, anti-dsDNA antibody, Scr, and BUN than the stable LN group (P<0.05). The LN, active LN, and stable LN groups had significantly lower levels of complement C3, complement C4, and serum albumin (Alb) than the control group (P<0.05). The active LN group had a significantly lower level of Alb than the stable LN group (P<0.05). The LN, active LN, and stable LN groups had significantly lower plasma levels of miR-145 and miR-183 than the control group (P<0.01). The active LN group had significantly lower plasma levels of miR-145 and miR-183 than the stable LN group (P<0.01). The children with difference types of LN had significantly lower plasma levels of miR-145 and miR-183 than the control group (P<0.01), and the type V-VI group and the type IV group had significantly lower plasma levels of miR-145 and miR-183 than the type II-III group (P<0.01). The ROC curve analysis showed that the optimal cut-off values of plasma miR-145, miR-183, and anti-dsDNA antibody were 1.05, 0.62, and 186.30 IU/mL respectively, in the diagnosis of LN, and the combination of these three indices had the largest area under the ROC curve of 0.896 (95%CI: 0.835-0.955), with a sensitivity of 90.5% and a specificity of 84.2%. In the children with LN, the plasma levels of miR-145 and miR-183 were negatively correlated with SLEDAI score, anti-dsDNA antibody, Scr, and BUN (P<0.05) and were positively correlated with complement C3, complement C4, and Alb (P<0.05). CONCLUSIONS There are significant reductions in the expression levels of miR-145 and miR-183 in plasma in children with LN, which are correlated with the activity level and pathological typing of LN. Combined measurement of miR-145, miR-183, and anti-dsDNA antibody has a high value in the diagnosis of LN.
Biomarkers ; Child ; Complement C4 ; Humans ; Lupus Nephritis ; genetics ; MicroRNAs ; genetics ; ROC Curve

BACKGROUND & OBJECTIVE:Glutathione is involved in cellular protection against radiation damage and drug detoxification.This study was to investigate the circadian variation of plasma cortisol and whole blood reduced glutathione(GSH)levels in nasopharyngeal carcinoma (NPC)patients to provide references for chronotherapy for NPC.METHODS:A total of 13 NPC patients and 14 healthy volunteers were involved.Peripheral venous blood was sampled every 4 h during one 24hour period starting at 12:00 am.The plasma cortisol concentration was determined by radioimmunoassay;the GSH concentration was determined by high performance liquid chromatography.RESUL-TS:Plasma cortisol levels of both groups displayed clear and similar circadian rhIythms.The cortisot level peaked in both groups in the morning and was the lowest at mid-night.In both groups,GSH concentrations showed significant differences according to sampling time(ANOVA for Repeated Measures,F=5.18,P=0.02).By cosinor analysis.the circadian variation of the GSH Ievel in NPC group was marginally statistically significant(Cosinor analysis,P=0.06)with the acrophase appeared at 05:02:the GSH Ievel in control group displayed an obvious circadian rhythm and the acrophase appeared at 07:44+01:56(P＜0.01).The rhythm-adjust mean value of glutathione was(19.60+1.11)nmol/mg protein in NPC group and(8.95,±0.46)nmol/mg protein in control group.CONCLUSIONS:The circadian rhythm of the plasma cortisol level is maintained in NPC patients,even in some patients at advanced stages.In NPC patients,GSH shows a trend of circadian variation which is similar to that in healthy controls.These results could be used to select special schedules for chrono-radiotherapy and chrono-chemotherapy for NPC patients.

Objective To compare the effectiveness of Defocus Incorporated Multiple Segments (DIMS) and High Aspheric Lenslet (HAL) defocus lenses in controlling myopia. Methods A total of 214 children and adolescents who were fitted with defocus lenses (DIMS or HAL) in our hospital from January to June 2023 were selected, including 100 cases in DIMS group and 114 cases in the HAL group. Changes in parameters such as spherical lens power, cylindrical lens power, spherical equivalent (SE), corneal curvature (K), axial length (AL), binocular accommodative response (BCC), positive/negative relative accommodation (PRA/NRA), distance/near latent phoria (DLP/NLP), and accommodative convergence to accommodation ratio (AC/A) were compared between the two groups before wearing and after wearing the lenses for 6 months, and the effectiveness of myopia control between DIMS and HAL was further compared. Results After wearing the lenses (DIMS or HAL) for 6 months, the average increase in axial length was (0.09±0.14) mm, the average increase in spherical lens power was (-0.11±0.23) D, the average increase in astigmatism was (-0.07±0.16) DC, and the average increase in spherical equivalent was (-0.15±0.24) D, with statistically significant differences observed for all these changes (t=9.24, -7.27, -6.60, -9.03, respectively, P < 0.001). Compared with parameters before wearing the lenses, statistically significant differences were noted after wearing for 6 months in spherical lens power, cylindrical lens power, SE, BCC, PRA, DLP, average K, and AL (P≤0.001), except for NRA (t=-0.12, P>0.05), NLP (t=-0.28, P>0.05), and AC/A (t=0.33, P>0.05). The differences in parameters between the two groups after wearing the lenses for 6 months showed a statistically significant difference only in the AL difference (t=4.38, P≤0.001), with no statistically significant differences observed for other parameter differences (P>0.05). Conclusion Wearing defocus lenses has a positive effect on controlling myopia, delaying axial length elongation, and maintaining binocular visual function balance.

OBJECTIVETo observe the injury on micro-skin induced by a self designed micro-skin machine.

METHODSMicro-skin was produced either with the machine or by hand. Cells at the edge of micro-skin were observed by transmission electron microscope. succinic dehydrogenase activity in supernatant of cultivated cells was analyzed, and the cell proliferation of micro-skin was assessed by (3)H-TdR. Twenty patients were enrolled in the study for the observation of the wound healing time between the two groups of micro-skin after being grafted.

RESULTSTransmission electron microscope examination revealed that the cellular injury at the edge of the micro-skin in machine-made group was mild compared with that in man-made group. (3)H-TdR rate was elevated but the activity of succinic dehydrogenase in the supernatant of cultured cells decreased in supernatant of cultured cells of machine produced micro-skin. Wound healing time was shortened in machine made group. (P < 0.05).

CONCLUSIONThe cellular injury at the edge of micro-skin in the machine made group was mild when compared with that in the man-made group with cell proliferation accelerated and wound healing time shortened.

Burns ; surgery ; Cell Division ; Epithelium ; pathology ; Humans ; Microscopy, Electron ; Skin ; ultrastructure ; Skin Transplantation ; methods ; Wound Healing