Stem Cells ; Toxicology ; methods

Background Most anti-inflammation eyedrops are limited in clinical application owing to multiple adverse effects.A novel peptide GC31 derived from human thrombomodulin has a natural anti-inflammatory activity.Compared with conventional anti-inflammatory eyedrops,GC31 possesses more advantages and potential clinical transforming value.However,relevant study is still lack.Objective The purpose of this study was to evaluate the anti-inflammatory effect of GC31 and the possible mechanisms.Methods Sixty SPF male Wistar rats aged 8-10 weeks were randomized into 6 groups using randomized number table.Non-specific keratitis models were established in 40 rats by intrastromal injection of 10 μl of lipopolysaccharide (LPS) dissolved in PBS.Different doses of GC31 (125 μg or 250 μg) or dexamethason soluble in PBS were sunconjunctically injected in the experimental eyes respectively in the low dose GC31 group,high dose of GC31 group and the dexamethason group,and 10 μl of PBS was used in the same way in the PBS control group.No drug was injected in the model group,and the normal rats were employed as the blank control group.The corneas were examined by slit lamp microscope and were scored based on the criteria of Anand 24 hours after injection.Then the corneas were collected for histopathological examination.Expression of nuclear factor-κB (NF-κB) p65 in the corneas was detected using immunochemistry.Expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) proteins were assayed using ELISA.Real-time PCR was used to detect the expressions of IL-6 mRNA and TNF-α mRNA.The use and care of the experimental animals followed Regulation for the Administration of Affair Concerning Experiment animals by State Science and Techonology Commission.Results A significant difference was seen in the ocular inflammatory scores among the six groups (F =301.238,P =0.000).The inflammatory scores were significantly lower in the high dose of GC31 group than those in the model group (1.85 ± 0.36 versus 2.90± 0.43) (t' =-5.144,P =0.000) ; and the scores in the dexamethason group was lower than those in the high dose of GC31 group(t' =-3.931,P=0.000).Infiltration of inflammatory cells in corneal tissue was milder in the high dose of GC31 and the dexamethason group compared with the model group.The positive response for NF-κB p65 was obviously weaker in the rat corneas in the low and high dose of GC31 groups and the dexamethason group in comparison with the model group.The contents of IL-6 and TNF-α proteins in the corneas were significantly reduced in the low and high dose of GC31 group and the dexamethason group compared with the model group (low dose group:t=-2.626,P=0.009;t'=-2.310,P=0.017.high dose group:t =-3.361,P=0.001 ;t'=-3.151,P=0.002),and the contents of IL-6 and TNF-α proteins in the dexamethason group were lower than those in the high dose of GC31 group (t=-3.361,P=0.001;t'=-3.360,P=0.000).In addition,the expression trend and compared results of IL-6 mRNA and TNF-α mRNA among the groups were similar to those of the IL-6 and TNF-α proteins (all at P＜0.01).Conclusions GC31 suppresses LPS-induced corneal inflammation response by downregulating the expression of inflammatory eytokines.The effect is more dominant in the doses of 250 μg than that in the doses of 125 μg.

AIM To explore the radioprotective function of Xihuang Capsules (Bovis Calculus artijactus,Moschus,Olibanum,Myrrha) in patients with acute radiation-induced oral mucositis and its mechanisms.METHODS Eighty patients with nasopharyngeal carcinoma undergone radiotherapy were randomized into two groups:treatment group (radiation plus Xihuang Capsules) and control group (radiation alone).The comparison of two groups in the onset and the tolerated dose of mucositis was made according to the acute radiation injury classification standard.The levels of TNF-α and IL-6 in saliva were detected by ELISA.RESULTS After the treatment with Xihuang Capsules,the onset of oral mucositis delayed significantly and the tolerated dose elevated markedly comparing with the control group (t =2.180,12.930,P ＜ 0.05).The morbidity rate of Ⅲ-N-degree of oral mucositis in the treatment group was lower than that of the control group at the dosage of 40 Gy and 70 Gy,respectively (Z =3.661,4.270,P ＜0.01).Furthermore,there was no difference in the levels of TNF-α and IL-6 in the pre-treatment in the two groups (t =1.010,1.469,P ＞ 0.05).With the increase in radiant dose,the levels of TNF-α and IL-6 in the two groups both elevated dramatically,and peaked at the dose of 40 Gy,but both levels in the treatment group were lower than those of the control group (t =8.305,6.069,P ＜ 0.05).When DT =70 Gy,the levels of TNF-α and IL-6 were lower than pre-radiation,and the levels of TNF-α and IL-6 in the treatment group were lower than those in the radiation alone group (t =3.835,2.488,P ＜ 0.05).CONCLUSION Xihuang Capsules can delay and reduce acute radiation-induced oral mucositis and improve radiation tolerated dose,so it may involve the release of TNF-oα and IL-6 in saliva.

Objective To investigate the effect of tramadol combined with sufentanil on postop-erative analgesia in patients received upper-abdominal surgery.Methods One hundred and fifty pa-tients scheduled for selective upper-abdominal operation were randomly divided into five groups (n=30 each):Tramadol group (T);large dose sufentanil combined small dose tramadol group (TS1 );balanced tramadol and sufentani group (TS2 );small dose sufentanil combied large dose tramadol group (TS3);sufentanil group (S).Postoprative VAS scores were recorded respectively at 1,4,8, 12,24,36,48 h when be in quiet and turning 90 degree,Ramsay sedation score,analgesia pump pressing times and side effects were also recorded.Results VAS scores in group TS2,group TS3 and group S at each time point both be in quiet and turning 90 degree were significantly lower than that of group T and group TS1 (P <0.05).Group S sedation score significantly higher than group T (P <0.05).No significant difference in other groups.Conclusion Balanced tramadol and sufentanil group has best analgesic effect and least side effects.With the increasing doses of sufentanil,its analgesic effect was not obvious enhancement.

Objective To investigate the effect of early enteral nutrition on prognosis of severe acute pancreatitis (SAP) .Methods Fifty-eight patients with SAP were randomly divided into early enteral nutrition group (experimental group ,n=30) and total par-enteral nutrition(TPN) group(control group ,n=28) .The experimental group was feed by Nose-jejunum nutrition tube and the con-trol group were supported with TPN through central vena .Compared the differences in complication incidence rate ,infection rate , mortality rate ,length of hospital stay and costs between two groups .Results The incidence rate of complications ,infection rate , length of hospital stay and hospital costs in experimental group were lower than control group ,the difference was statistically sig-nificant(P<0 .05) .The mortality in experimental group was lower than control group ,but the difference was not statistically sig-nificant(P>0 .05) .Conclusion Early enteral nutrition support therapy can improve the nutritional status of patients with SAP ,re-duce the incidence of complications ,infection ,length of hospital stay and hospital costs .

Objective To isolate and culture sweat gland epithelial cells in vitro,and to study the effects of acetylcholine (ACh) on intracellular flee calcium concentration ([Ca~(2+)]i) of cultured sweat gland epithelial cells.Methods Sweat glands epithelial cells were collected by enzymatic digestion.After ACh was added to the primary and first passage cells,[Ca~(2+)]i was examined using confocal laser scanning microscopy (CLSM) and the Ca~(2+) sensitive dye Fura 3/AM.Results The primary and first passage epithe- lial cells grew well.After ACh was added,opening of the calcium channel and significant [Ca~(2+)]i increase were observed when the primary and first passage cells were incubated with high concentration of calcium (2 mmol/L);no significant [Ca~(2+)]i increase was observed in those cultured without calcium.Conclusion Upon stimulation with ACh,calcium channels of cultured primary and first passage sweat gland epithelial cells would open,influx of extracellular Ca~(2+) occurred,which resulted in an increase of [Ca~(2+)]i.Extracellular bound calcium was therefore converted into intracellular free calcium.

Objective To compare the growth of Plasmodium vivax in Anopheles sinensis(An.s) and Anopheles anthropophagus(An.a) during the periods of malaria clinic attack and diapause. Methods The blood samples of patients during the clinic attack and diapause of vivax malaria patients in the vivax epidemic area in China were collected, feeding the mosquitoes of An.s and An.a by using the artificial membrane feeding system in vivo in the lab, and the mosquitoes were dissected during the day 7-9th and 14th after the infection and the oocysts and sporozoites in the stomach and gland of the mosquitoes were counted, respectively. Results The oocyst positive rate in An.s fed by Plasmodium vivax during the fever stage was lower than that in non-fever stage, the sporozoite positive rate in An.a fed by Plasmodium vivax during the fever stage was lower than that in non-fever stage. The positive mosquito rate with oocyst and sporozoite infected by Plasmodium vivax in the fever stage to An.s and An.a were lower than those in non-fever stage. The infective sporozoite intensity of An.s fed by Plasmodium vivax in the fever stage was lower than that in non-fever stage, but the reverse result was found to An.a. Conclusion There is a significant difference between the periods of malaria clinic attack and diapause of the oocyst and sporozoite infection to An.s and An.a.

Objective:To explore the preventive function for sweating gustatory syndrome and esthetic effect of restoring parotid gland area defect by using platysma flap. Methods: Nine patients who underwent parotidectomy and restored parotid gland area defect with platysma flap were analyzed retrospectively in this study. Observed whether there was an incidence of subjective sweating gustatory syndrome and introcession in the operation region. Results: There was no incidence of subjective sweating gustatory syndrome. Moreover, introcession in the operation region was not obvious. Conclusion: Restoring parotid defects with platysma flap can reduce introcession in the operation region. Application of platysma flap as an interpositional barrier between the skin flap and the parotid bed after parotidectomy can reduce the incidence of sweating gustatory syndrome.

ObjectiveTo build the computer-aided diagnosis system of left ventricular diastolic function for echocardiography,to make the evaluation process of left ventricular diastolic function for echocardiography more convenient,efficient,standardized and to obtain more accurate,practical,objective and reliable diagnostic results.MethodsFirst,the system was developed by the Macromedia Flash Professional referred to the newest guidelines and suggestions published by the American Society of Echocardiography(ASE) in 2009.Second,the program of left ventricular diastolic function in the computer- aided diagnosis system was designed,debugged and improved according to feedback information frompractical applications.ResultsThe diagnostic system can work simultaneously with echocardiography.If assessment parameters of the newest guidelines and suggestions,published by the American Society of Echocardiography in 2009,are imported to the system,then it can give the diagnosis of the change about the left ventricle filling pressures and score the left ventricular diastolic function quickly and automatically.The system will export standard report with both pictures and text.ConclusionsAccording to our experiment,the computer-aided diagnosis system is able to make the evaluation process of left ventricular diastolic function much more standardized and simplified,which guarantee the objectivity and reliability of the evaluation results.Consequently,it will be able to apply to clinical practice in the future.

Objective To evaluate the clinical significance of intercellular space diameters (ISD)of squamous epithelium by light microscopy (LM) in lower esophagus of erosive reflux esophagitis (ERD),non-erosive reflux disease ( NERD), Barrett esophagus (BE) and healthy controls. Methods A total of 21 ERD and 21 NERD patients with reflux symptoms and confirmed with 24-hour esophageal pH monitoring, 13 BE patients diagnosed by gastroscopy and biopsy, and 20 other healthy controls were enrolled in the study.Samples of ERD, NERD and control group were collected at 2 cm above dentate line, and made HE slides in the conventional way. Images for measurement of ICS were acquired with oil lens ( × 1000). ICS of squamous epithelium was quantitatively measured by computer-assisted morphometry. Ten cells were taken for each sample, 10 consecutive ISD for each cell, i.e. 100 ISD for each subject. Mean ISD was calculated.Results Mean ISDs by LM in control, BE, ERD, and NERD groups were 0. 59, 0. 99, 1.29 and 1.06 μm, respectively. The mean ISDs in BE, ERD, and NERD group were much greater than that in control (P＜0. 05). The mean, maximal and minimal ISDs of group ERD were greater than those of NERD and BE (P = 0. 000). However, the ISDs of NERD and BE are of no significant difference ( P ＞ 0. 05 ). The cut-off value of mean ISD for diagnosis of gastro-esophageal reflux disease (GERD) was 0. 85 μm. Diagnostic sensitivity and specificity for ERD, NERD and BE were 89. 1% and 100. 0%, with reference to clinical symptoms, endoscopy and ISDs above the cut-off value. Conclusion Larger ISDs in lower esophagus by using LM will be found in all subgroups of GERD, including ERD, NERD and BE. Increased ISDs may be one of the markers for diagnosis of ERD, NERD and BE.