Background Cytokines play a crucial role in mediating immune tolerance or immune rejection of corneal transplantation.However,the study on the expression of cytokines in corneal graft is seldom.ObjectiveThe purpose of the study is to investigate the change of cytokine expression in allografts at different time points after corneal transplantation.Methods BALB/c and C57BL/6 mice aged 6 to 8-week old were used to establish autologous and allografts keratoplasty models.BALB/c mouse was use as donor and receipt in autologous group,and the cornea of C57BL/6 mouse was used to graft on the BALB/c mouse in the allografts group.The graft inflammation was clinically scored,and graft inflammatory scores of ≥5 or opaciflcation scores of ≥2 were identified as rejection.BALB/c mice were randomized into normal control group(3 mice)and allografts group(15 mice).Reverse transcriptPCR (RT-PCR) was used to detect the expression of interleukin-4 (IL-4) mRNA,interferon-γ (IFN-γ) mRNA,IL-10mRNA and tumor necrosis factor(TNF-α) mRNA in graft 6 hours and 1 day,3,7,14 days after operation.Results The corneal graft opacification score was ＜2 and inflammatory score was ＜5 in the 10 mice with autologous keratoplasty until 60 days with the survival rate 100％.The edema,opacification and new blood vessel were seen in the BALB/c mice received allografts keratoplasty.The inflammatory score was ≥ 5 and the opacification score was ≥2 24 days after surgery with the rejection rate 100％,in the allografts group,and the graft survival time was (17.80±4.66)days.RT-PCR showed that IL-4 and IFN-γ were positively expressed,and IL-10 and TNF-α were absently expressed in normal mouse cornea.In the allografts keratoplasty mice,positive responses for IL-4,IFN-γ and IL-10 were found in 6hours after operation,but TNF-α was absent.From 1 day through 3 days after operation,the expressions of IL-4,IFN-γand TNF-α were enhanced but IL-10 was disappeared in the graft.IL-10,IFN-γ and TNF-α were expressed till the 7th day,but on the 14th day,only IL-10 was detected in graft in the allografts keratoplasty mice.Conclusions TNF-αis a main factor among the variety of cytokines that may influent corneal allograft rejection locally.

Objective To observe the gonadotropin analogue ( GnRHa) Triptorelin of idiopathic central pre-cocious puberty(ICPP) girls′body mass index(BMI) and final adult height(FAH).Methods 120 patients with idi-opathic central precocious puberty cases were summarized and track the return visit ,which.of 62 cases with ICPP girls Triptorelin treatment,6,12,18,24,36 months of observation and treatment of bone age (BA),the growth rate(GV), BMI,predicted height ( PAH) ,and life-long high and up to lifelong high BMI .The untreated group was also observed the relevant indicators such as BMI ,and lifelong.Results After treatment,the increase in BA was less than that in age.The PAH has improved with the growth of the treatment regimens .After treatment PAH 12,18,24,36 months PAH respectively[(153.4 ±7.1)cm,(154.6 ±6.2)cm,(155.7 ±4.7)cm,(156.9 ±5.9) cm], both were statisti-cally significant(t=2.23,2.67,3.01,2.88,all P<0.01); BMI [(16.37 ±1.41)kg/m2,(16.99 ±1.21)kg/m2, (17.17 ±1.38)kg/m2,(17.02 ±1.42)kg/m2] between the two were not statistically significant (t=0.69,1.28, 0.89,1.11,all P>0.05).Lifelong treatment and control groups,respectively[(158.2 ±3.9)cm,(153.7 ±2.8) cm],and between the two there is a statistically significant (t=3.04,P<0.01).BMI were up to lifelong[(19.1 ± 1.9)kg/m2,(18.7 ±1.8) kg/m2] between the two was not statistically significant (t =0.35,P >0.05). Conclusion Triptorelin can effectively suppress idiopathic central precocious sexual characteristics and bone age growth and improve adult height .There is no significant effect on the BMI .

Objective: To isolate and identify the chemical constituents in the ethanol extract from the barks of Illicium brevistylum. Methods: The barks of I. brevistylum were extracted by 80% ethanol then partitioned by system solvents with different polarity. The ethyl acetate extract was separated on silica gel, Sephadex LH-20, and ODS columns. The isolated compounds were identified by physicochemical properties and spectral analyses. Results: Twelve compounds were isolated and purified from the ethyl acetate extract of I. brevistylum and the structures were identified as quercetin 3-O-β-xylopyranoside (1), quercetin (2), dihydro kaempferol (3), kaempferol (4), wulingzhic acid V (5), catechin (6), 3, 5, 7-trihydroxylchromone-3-O-α-L-arabinopyranoside (7), majucin (8), anislactone B (9), 7-O-acetylanislactone B (10), dunnianolide C (11), and taxifolin-3-O-β-xylopyranoside (12). Conclusion: Compounds 5 and 7 are obtained from the plants in Illiciaceae for the first time and compounds 1, 6, and 8-11 are firstly isolated from I. brevistylum.

Background Phlyctenular ophthalmia is a delayed hypersensitivity reaction to some microprotein and affected mainly by adolescent in high incidence.Objective This study was to investigate the microscopy findings of phlyctenular ophthalmia and evaluate the histological changes by laser scanning confocal microscope.Methods Twenty-nine eyes suffered from phlyctenular ophthalmia and twenty normal eyes were examined using laser scanning confocal microscope.The pictures were taken by a CCD camera.All the cases had initially chest X-ray,tuberculin test,bacterial and mycobacteria culture.Results Dendritic and inflammatory cells were increased and concentrated in conjunctiva,and epithelial cells were deformed and squamatizated.The capillaries engorged and the goblet cells were injured.The corneoscleral Vogt meshing of the phlyctenular keratitis was obscured and dendritic cells were intruded into the corneas.The corneal epithelium of phlyctenular keratitis was absent and the subepithelial nerve plexus were bended and fractured,and the dendritic and inflammatory cells were intruded.Scarring of corneal stroma was seen under the laser scanning confocal microscope.Conclusions Laser scanning confocal microscopy is valuable for basic research and clinical diagnosis of phlyctenular ophthalmia.

Objective To investigate the effect of high mobility group box-1 protein (HMGB1) on cytokine expression in splenic dendritic cell (DCs). Method DCs isolated from the spleens of male Wistar rats were seed-ed on 96-well (1×10s cells/well) cell culture plates, and the cells were stimulated with HMGB1 for various length of time or in different concentrations. (1) The time-dependent response between HMGB1 and tumor necrosis factor-α(TNF-α) as well as interleukin-12 (IL-12) gene/protein expressions: 24 wells of DCs were dividedly into six groups including 24 h-normal controls (n=4), 48 h-normal controls (n=4), 72 h-normal controls (n=4), and 24 h-HMGB1 treated group (n=4), 48 h-HMGB1 treated group (n=4) as well as 72 h-HMGB1 treated group (n=4), respectively. Among three HMGB1-treated groups, DCs were stiraulated by 1 μg/mL HMGB1. DCs were denatured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvested to determine Il-12 as well as TNF-α protein levels. (2) The dose-dependent response between HMGB1 and TNF-α as well as IL-12 gene/protein expressions: 16 wells of DCs were dividedly into four groups in-cludingnormal controls (n=4), 0.1 μg/mL HMGB1 treated group (n=4), 1 μg/mL HMGB1 treated group (n =4), and 10 μg/mL HMGB1 treated group (n=4), respectively. After stimulated for 48 h, DCs were dena-tured in cell culture plates to determine gene expression of IL-12 as well as TNF-α, and supernatants were harvest-ed to determine IL-12 as well as TNF-α protein levels. Total RNA was extracted from cells using the single-step technique of acid guanidinium thiocyanate-chloroform extraction according to the manufacturer' s instruction (Promega, Madison, WI). mRNA for TNF-α and IL-12 were quantified by SYBR Green two-step, real-time re-verse transeription-polymerase chain reaction taking glyceraldehyde-3-phesphate dehydrogenase (GAPDH) as an internal standard. Levels of IL-12 and TNF-α in cell culture supernatants were determined with ELISA, strictly fol-lowing the protocols provided by the manufacturer. Data were analyzed with a one-way ANOVA. A P-values <0.05 were considered statistically significant. Results After stimulation with 1 μg/mL HMGB1, IL-12 and TNF-α protein and gene expressions in rat splenic DCs were markedly up-regulated at 24 h to 72 h (P<0.05 or P<0.01), and the expression levels of IL-12 and TNF-α peaked at 48 h (P<0.01). When DCs were cultured in the presence of 0.1 μg/mL, 1 μg/mL, and 10 μg/ml HMGBI for 48 h, expressions of IL-12 and TNF-α were also significantly up-regulated (P<0.01), and values of these cytokines were highest in 1 μg/mL HMGB1-treated group (P<0.01). Conclusions These data suggest that HMGB1 appears to be a potential immunostimulatory signal that induced DC maturation, and HMGB1 stimulation can result in marked up-regulation of IL-12 as well as TNF-α synthesis and release in splenic DCs.

Objective To prepare quercetin liposomes and establish a method for determination of its entrapment efficiency.Methods The film dispersion-homogenizing method was used to prepare quercetin liposomes.The formulation was optimized on the basis of orthogonal design and its entrapment efficiency was performed by the protamine sedimentation method.Results The optimal conditions were found to be cholesterol-egg phospholipid=1:3,quercetin-vehicle = 1:40,homogenization pressure 103.4 MPa for three times.The average entrapment efficiency of the optimized nano-liposomes was 92.1%.Conclusion The film dispersion-homogenizing method could be used to prepare quercetin liposomes.The protamine sedimentation method is convenient,accurate,and suitable for the determination of the entrapment effi- ciency of quercetin liposomes.

Partial nature of "promoting blood circulation and dieresis" of Salvia Miltiorrhizain was initially demonstrated by investigating the regulation effect of AQP2 expression in kidney of trauma blood stasis model rats with the Salvia Miltiorrhizain so as to provide guidance for its clinical deployment of administration. Random allocation was taken to averagely divide 30 SD rats into two groups: 10 rats in normal group and 20 rats in blood stasis syndrome group. Trauma blood stasis rat model was established by quantitatively beating. Then the rat model group was divided into model group and salvia group. After 7 days of treatment, the rat kidney AQP2 expression was detected, the content of urine AQP2 was compared and the damaged local muscle and kidney pathological changes were observed by immunohistochemical method and western blot method. Compared with that of the normal group, rats in model group had inflammatory cells infiltration, blood stasis and edema of the injured local muscles and up-regulated AQP2 expression, decreasing urinary output, and kidney tissues blood stasis and edema (P < 0.05). On the other hand, compared with that of the model group, those parameters of rats in salvia group were all decreasing except urine output (P < 0.05). Such result indicated that Salvia Miltiorrhiza can reduce trauma blood stasis rat content of urine AQP2 and down-regulated AQP2 expression in kidney tissue, so as to reduce the reabsorption of water by renal tubular and increase urine output. The promoting blood circulation effect of Salvia Miltiorrhizain can alleviate the degree of the damaged tissue edema and encourage urine drainage. This therapy is closely related to the effect of regulating AQP2 in kidney by salvia, so the purpose of this study by verifying "promoting blood circulation and diuresis" as the mechanism for the regulation effect of the salvia on AQP2 expression.
Animals ; Aquaporin 2 ; genetics ; metabolism ; Blood Circulation ; drug effects ; Diuresis ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Kidney ; blood supply ; drug effects ; metabolism ; physiopathology ; Kidney Diseases ; drug therapy ; genetics ; metabolism ; physiopathology ; Male ; Rats ; Salvia miltiorrhiza ; chemistry

ATP Binding Cassette Transporter, Sub-Family B ; metabolism ; Apoptosis ; genetics ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; pathology ; Genes, MDR ; Humans ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Telomerase ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; metabolism ; Vault Ribonucleoprotein Particles ; metabolism

Objective:To find the reasons for the low dissolution of clarithromycin tablets and study the interference from filter film adsorption at various time points to explore the appropriate processing approach for dissolution solution of clarithromycin tablets. Meth-ods:Clarithromycin tablets from two different manufacturers were used. The dissolution solution was prepared according to Japanese Orange Book. The dissolution was determined after different processing and the adsorption rate of the filter film was calculated. Re-sults:Totally 14 kinds of filter films were tested with different adsorption for clarithromycin, and the adsorption rate of some kinds of filter films exceeded the prescribed limit. Conclusion:The absorption rate of filter films for clarithromycin can be decreased by boiling the films and using American membranes. The interference from filter film adsorption can be reduced and inhibited by rejecting the first filtrate above 5ml or centrifuging dissolution solution.

Objective To assess set-up errors measured with kilovoltage cone-beam CT (KV-CBCT), and the impact of online corrections on margins required to account for set-up variability during IMRT for patients with prostate cancer. Methods Seven patients with prostate cancer undergoing IMRT were enrolled onto the study. The KV-CBCT scans were acquired at least twice weekly. After initial set-up using the skin marks, a CBCT scan was acquired and registered with the planning CT to determine the setup errors using an auto grey-scale registration software. Corrections would be made by moving the table if the setup errors were considered clinically significant ( i. e. , ＞ 2 mm). A second CBCT scan was acquired immediately after the corrections to evaluate the residual error. PTV margins were derived to account for the measured set-up errors and residual errors determined for this group of patients. Results 197 KV-CBCT images in total were acquired. The random and systematic positioning errors and calculated PTV margins without correction in mm were:a) Lateral 3. 1,2. 1,9. 3;b) Longitudinal 1.5, 1.8, 5. 1 ;c) Vertical 4. 2,3.7, 13.0. The random and systematic positioning errors and calculated PTV margin with correction in mm were:a) Lateral 1.1,0. 9, 3.4;b) Longitudinal 0. 7, 1.1, 2. 5;c) Vertical 1.1, 1.3, 3.7. Conclusions With the guidance of online KV-CBCT, set-up errors could be reduced significantly for patients with prostate cancer receiving IMRT. The margin required after online CBCT correction for the patients enrolled in the study would be appoximatively 3-4 mm.