Objective:To explore the establishment and evaluation method of recurrent infection model of herpes simplex keratitis (HSK).Methods:To determine the optimal duration of the ultraviolet light irradiation, 12 healthy BALB/c mice were randomized into 3 minutes group, 2 minutes and 45 seconds group, and 2 minutes and 30 seconds group based on the duration of the ultraviolet light irradiation according to random number table method, with 4 mice in each group.Another 72 healthy BALB/c mice were randomized into blank control group, model group and recurrence group according to random number table method, with 24 mice in each group.The 72 mice were scratched a # symbol on their right corneas with scalpel.Then the eyes in the blank control group were treated with 5 μl of normal saline solution, while the eyes in the model group and recurrence group were treated with 5 μl of herpes simplex virus I (HSV-1) suspension.All the 72 mice were not intraperitoneally injected with HSV-1 xenogeneic serum antibody.Five weeks after the initial infection, the mice in the recurrence group were irradiated at ultraviolet B-302 nm for the optimal duration.The feasibility of establishing HSK recurrence model induced by this method was evaluated by the score of ocular surface symptoms, and the cytopathic effect (CPE) lesion analysis of Vero cells cultured in corneal wiping fluid from the mice in the model group and recurrence group.The use and care of the animals complied with the Statement of the Association for Research in Vision and Ophthalmology (ARVO). The study protocol was approved by the Ethics Committee of Shanghai University of Traditional Chinese Medicine (No.PZSHUTCM190222039).Results:The optimal duration of the ultraviolet light irradiation was 2 minutes and 45 seconds.Within one week, HSK syndromes appeared in the mice injected with HSV-1 virus in the model group and recurrence group, and then gradually disappeared after one week.Before inducing recurrence, slit lamp examination was performed, and there was no spontaneous recurrence of HSK in the model group and recurrence group.The mice in the recurrence group relapsed within one week after ultraviolet light irradiation.The symptom scores of ocular surface lesions in the blank control group, model group and recurrence group were 0.333±0.471, 1.500±0.764 and 2.667±0.943 at one day after ultraviolet light irradiation, 0.000±0.000, 0.833±0.373 and 5.167±2.267 at three days after ultraviolet light irradiation, and 0.167±0.373, 1.000±0.577 and 3.000±1.155 at seven days after ultraviolet light irradiation, respectively.The symptom score of ocular surface lesions was higher in the recurrence group than the blank group and the model group at three days after ultraviolet light irradiation, and the differences were statistically significant (both at P<0.05). Morphological changes such as floating and gathering were found in the Vero cells after being cultured in corneal wiping fluid.The positive rate of CPE lesion was 71% (17/24) in the recurrence group, which was significantly higher than 8% (2/24) in the model group, and the difference was statistically significant ( P<0.01). Combining the two indicators, the success rate of recurrence in established models could reach 71%. Conclusions:Ultraviolet light irradiation can successfully induce the recurrence of viral keratitis in HSK mice without injection of neutralizing serum antibody.

Objective To investigate the effect of intravenous thrombolysis with different doses of alteplase on neurological function in patients with acute ischemic stroke.Methods 120 patients with acute ischemic stroke were selected as the research subjects.According to the admission time,the patients were divided into the observation group and the control group,60 patients in each group.The observation group was treated with low dose alteplase(0.6mg/kg),the control group was given standard dose(0.9mg/kg).The clinical curative effect,incidence rate of adverse event,NIHSS score and mRS score before and after treatment were observed and recorded in the two groups.Results Before treatment,there was no significant difference between the two groups in NIHSS score(t=0.288 0,P=0.773 9).After treatment for 3 days,the NIHSS score of the control group decreased compared with before treatment,and was lower than that of the observation group at the same period,there was significant difference between the two groups(t=0.778 5,P=0.000 0).Before treatment,the mRS score between the two groups had no siginificant difference(t=0.801 0,P=0.424 7).After treatment for 90 days,the mRS score of the two groups decreased,the difference was statistically significant(t=6.811 5,P=0.000 0).After treatment,1 patient with cerebral hemorrhage was observed in the observation group,while 3 cases in the control group,the difference between the two groups was not statistically significant(χ2=1.034 5,P=0.309 1).The mortality of the observation group was 11.47％,which of the control group was 1.67％,there was significant difference between the two groups(χ2=4.821 4,P=0.028 1).The good prognosis rate of the observation group was 45％,which of the control group was 68.33％,the difference was significant between the two groups (χ2=6.651 6,P=0.009 9).Conclusion Standard dose of alteplase in the treatment of acute ischemic stroke has better efficacy than low-dose alteplase,it has improved conditions of neurological deficits in patients,better recovery of neurological function,the prognosis is good,it can be widely recommended.

Objective To investigate the hypothesis that food allergy in infants may be associated with variation in their intestinal microflora. The formation of intestinal microflora in healthy infants and changes in food allergic infants were detected.Methods 16S rRNA gene sequences specific for bifidobacterium, lactobacillus and escherichia coli in fecal were quantitatively detected by real-time PCR. The three fecal floras were assessed in 71 healthy infants and 100 infants with food allergy. Results After birth,there were bifidobacteria colonized in infantile intestine,then the number increased rapidly up to 5 times at the sixth month, which was always the preponderant flora. Lactobacilli was also presented in infantile intestine 1 month after birth and augment gradually. The number of Escherichia coli was less than bifidobacteria and lactobacilli and appeared to decline during the early infants. The number of bifidobacteria and lactobacilli in the infants with food allergy were markedly less than that in the healthy infants, but escherichia coli was significantly more than that in the healthy infants.Conclusions During the first year of life,the intestinal microflora in infants is in a developing process. Compared with the healthy infants,bifidobacteria and lactobacilli decrease, but escherichia coli increase in the food allergic infants.These results indicate that the probiotics may be benefit to the prevention and treatment of food allergy.

Endophytic fungi played an important role in the growth of its host plant. To investigate the mycorrhizal characteristics and the distribution of fungi in the root, an endangered wild plant-Dysosma versipellis was collected and observed by electron microscope. The results showed that the host was closely associated with endophytic fungi. The fungi were mainly distributed in the epidermis and cortex. The aseptate and septate fungi with swollen hyphae were observed in some cell of the cortex. The result provides a reference for the study of mycorrhizal structure of Dysosma genus and the interaction between the fungi and its host.
Berberidaceae ; microbiology ; ultrastructure ; Endangered Species ; Endophytes ; physiology ; ultrastructure ; Fungi ; physiology ; ultrastructure ; Microscopy, Electron ; Plant Roots ; microbiology ; ultrastructure

objective To explore the role of cytokines and dendritic cells (DCs) in rat high-risk corneal allograft survival pro- longed by superantigen Staphylococcal Enterotoxin B (SEB) and to compare the different effects between SEB and glucocorticoid. Design Experimental study.Participants Fisher 344 and Lewis rats.Methods The Fisher 344 and Lewis inbred rats were used for high-risk penetrating keratoplasty model.All of the Lewis rat recipients were divided into three groups by blinded fashion.GroupⅠand GroupⅡrats were injected intraperitoneally with 0.2ml saline buffer or SEB (75?g/ml) respectively at 4-day intervals on three occasions before transplantation.GroupⅢrats were injected subconjunctivally with 0.1ml dexamethasone (1mg/ml) daily from the first day after surgery for 2 weeks.The allograft survival was examined under slit-lamp.The concentration of interleukin IL-1?,IL-2,IL-4,IL-5,IL-6, IL-10,TNF-?in rat aqueous humor and peripheral blood were measured by liquichip and the cytokine and CD11c,CD80,MHC-Ⅱex- pression in corneal grafts were examined by immunohistochemestry staining.Main Octcome Measures Mean survival time of the allo- grafts,the level of cytokines in aqueous humor,peripheral blood and corneal grafts.Results Compared with group control,the grafts mean survival time was delayed about 3.8d in group SEB (P=0.00) and 7.1d in group dexamethasone(P=0.01).But there was no signifi- cant difference between group SEB and dexamethone (P=0.26).Liquichip test showed that the level of IL-1?in aqueous humor was re- duced and IL-4,IL-6 and IFN-?,ascended.Only IFN-?,and IL-6 could be found in peripheral blood,and the changing shift of them was similar to that in aqueous humor.The immunohistochemistry staining showed that the expression of IL-2 in rat corneal grafts was signif- icantly decreased but IL-4,IL-6 and IL-10 elevated.The expression of DCs in group SEB was similar to that in group control,which was elevated after keratoplasty,but the phenotype of DCs was not the same.There were predominantly mature DCs in group control while immature DCs in group SEB.Conclusions The immunological inhibiting effect of SEB is same to glucocorticoid,but the mecha- nism is different.SEB can modulate immune response,which might induce immune inhibition via the local production of cytokines and effect on DCs maturation involving corneal graft rejection prevention.

OBJECTIVETo evaluate the clinical effect and safety of dapoxetine in the treatment of premature ejaculation (PE).

METHODSWe randomly assigned 116 PE patients to receive dapoxetine on demand at 30 mg qd (dapoxetine group, n = 60, aged 23-49 years) or oral tamsulosin at 20 mg qd (control group, n = 56, aged 24-46 years). After 4 weeks of medication, we compared the clinical global impression of change (CGIC) , PE profile (PEP) scores, intravaginal ejaculation latency time (IELT) , and adverse reactions between the two groups of patients.

RESULTSCompared with the baseline, the IELT was remarkably prolonged after treatment both in the dapoxetine group ([0.86 ± 0.17] vs [4.32 ± 2.23] min, P < 0.05) and the control ([0.88 ± 0.15] vs [4.17 ± 2.26] min, P < 0.05), with no statistically significant difference between the two groups (P > 0. 05). The post-treatment rate of CGIC in the dapoxetine group had no statistically significant difference from that in the control (85.00% vs 82.14%, P > 0.05). In comparison with pre-treatment, the patients of both the dapoxetine and control groups showed dramatically improved scores after medication in perceived control over ejaculation (0.85 ± 0.23 vs 2.13 ± 0.97 and 0.88 ± 0.21 vs 2.06 ± 0.34, both P < 0.05), ejaculation-related personal distress (1.15 ± 0.64 vs 2.89 ± 0.26 and 1.19 ± 0.53 vs 2.82 ± 0.69, both P < 0.05), satisfaction with sexual intercourse (0.81 ± 0.33 vs 2.58 ± 0.37 and 0.79 ± 0.28 vs 2.45 ± 0.32, both P < 0.05), and ejaculation-related interpersonal difficulty (2.05 ± 0.61 vs 3.24 ± 0.35 and 2.03 ± 0.65 vs 3.18 ± 0.76, both P < 0.05), with no significant differences between the two groups (P > 0.05). The incidence of adverse reactions was significantly lower in the dapoxetine than in the control group (3.33% vs 30.36%, P < 0.05).

CONCLUSIONDapoxetine is effective for the treatment of PE, with its advantages of prolonging the intravaginal ejaculation latency time, improving the quality of sexual life, and low incidence of adverse reactions.

Adult ; Benzylamines ; administration & dosage ; therapeutic use ; Coitus ; Double-Blind Method ; Ejaculation ; Humans ; Male ; Middle Aged ; Naphthalenes ; administration & dosage ; therapeutic use ; Patient Satisfaction ; Premature Ejaculation ; drug therapy ; Serotonin Uptake Inhibitors ; administration & dosage ; therapeutic use ; Sexual Behavior ; Sulfonamides ; administration & dosage ; therapeutic use ; Treatment Outcome ; Young Adult

The aim of this research is to investigate the influence of microencapsulation on the expression of the oxidative stress genes and exogenous regulation of HepG2 cells. We compared the expression of hemeoxygenase-1 (HO-1) and glutathione S-transferases-A1 (GST-A1) in HepG2 cells under different culture conditions through real-time PCR. The effects of exogenous antioxidants on cell viability and albumin levels were also evaluated through MTT assay and ELISA assay. The results showed that after culturing for 6 and 16 days, the expression levels of HO-1 in encapsulated cells were approximately 4.9 and 3.1 times higher than that of monolayer cells at the same culture period; As for the expression levels of GST-A1, they were elevated to 11.2 and 33 times of monolayer cells (P < 0.05). Accordingly, we found that NAC at 5-10 mmol/L significantly increased the viability by 40%-70% and the biosynthetic function by 20%-30% in microencapsulated HepG2 cells (P < 0.05). GSH increased the viability of the encapsulated cells by 20%-55% and the biosynthetic function by 15% (P < 0.05). In conclusion, oxidative stress exists in the microcapsules and affects genes expression. Exogenous antioxidants can prevent the inhibition effects of oxidative stress on cellular growth.
Antioxidants ; pharmacology ; Cell Survival ; Gene Expression Regulation ; Glutathione Transferase ; metabolism ; Heme Oxygenase-1 ; metabolism ; Hep G2 Cells ; Humans ; Oxidative Stress

Age Distribution ; Aged ; China ; epidemiology ; Female ; Humans ; Male ; Middle Aged ; Sex Distribution ; Surveys and Questionnaires ; Tobacco Smoke Pollution ; statistics & numerical data

A combination technique of LC/ESR, LC/MS and spin trapping was used to identify and characterize the carbon-centered free radicals formed from lipoxygenase-catalyzed peroxidation of dihomo-γ-linolenic acid(DGLA). The spin trap, α-[4-pyridyl-1-oxide]-N-tert-butyl nitrone(POBN), could react with short-lived carbon-centered radicals to form relatively stable adducts. Based on the same retention time of POBN radical adduct in LC/UV/ESR and LC/MS, molecular weight of adduct could be confirmed and the structure of adduct could be confirmed by their LC/MS2 fragment pattern. The results showed that free radicals formed in lipoxygenase-catalyzed peroxidation of DGLA, including ~·C_7H_(13)O_2, ~·C_(10)H_(17)O_2 and ~·C_5H_(11), all stemmed from β-scission of DGLA alkoxyl radicals(8-, 11-, 15-LO~·). The results were helpful for further study of biological activities of these radicals in vivo.

Adult ; Aged ; Bronchoalveolar Lavage ; methods ; Critical Pathways ; Humans ; Male ; Middle Aged ; Pneumoconiosis ; therapy