Objective To investigate the interaction between NMDA receptor subunit 2B (NR2B) and metabotropic glutamate receptor 5 (mGluR5) in the spinal cord of morphine-tolerant rats. Methods Sixteen male SD rats weighing 220-280 g were randomly divided into 2 groups with 8 rats in each group: control group (C) and morphine group (M). In group M morphine 10μg was administered intrathecally (IT) twice a day for 7 consecutive days. In group C equal volume of normal saline was given instead of morphine. Tail flick latency (TFL) (a segment of tail 4-5 cm long was immersed in 52-53℃ hot water) was used to measure the antinocicepitve effect of morphine before and 30 min after morphine administration every morning. MPE (percentage of maximal possible effect) was calculated (pain threshold after drug administration - baseline pain threshold)/(10- baseline pain threshold)× 100% . The animals were killed at the next day after last IT administration and the dorsal half of L4-5 spinal cord was isolated and homogenized for determination of NR2B and mGluR5 protein expression by Western blot and CO-IP. Results MPE was significantly increased at 1-5 d of drug administration and returned to baseline at 7 d in group M as compared with group C. The mGluR5 protein expression was significantly up-regulated in group M as compared with group C but there was no significant difference in NR2B protein expression between the 2 groups. Conclusion There is interaction between NK2B and mGluR5 in the spinal cord of morphine-tolerant rats.

The Tongmai Yangxin pill (TMYX) has potential clinical effects on no-reflow (NR); however, the effective substances and mechanisms by which this occurs remain unclear. This study evaluates the cardioprotective effects and molecular mechanisms of TMYX against NR. We used a myocardial NR rat model (2 h after myocardial ischemia and 2 h after reperfusion) to confirm the effect and mechanism of action of TMYX in alleviating NR. In vitro studies in isolated coronary microvasculature of NR rats and in silico network pharmacology analyses were performed to reveal the underlying mechanisms of TMYX and determine the main components, targets, and pathways of TMYX, respectively. The experiment was approved by the Ethics Committee of Hunan University of Chinese Medicine (LLBH-202212160001). TMYX showed therapeutic effects on NR by improving cardiac structure and function, reducing NR, ischemic areas, and cardiomyocyte injury, and decreasing the content of cardiac troponin I (cTnI). Moreover, the mechanism of TMYX predicted by network pharmacology is related to the hypoxia inducible factor-1 (HIF-1), nuclear factor kappa-B (NF-κB), and tumor necrosis factor (TNF) signaling pathways. TMYX increased the expression of G protein-coupled estrogen receptor (GPER), phospho-extracellular signal-regulated kinase (p-ERK), and HIF-1α. In vitro, TMYX enhanced the diastolic function of coronary microvascular cells; however, this effect was inhibited by GPER inhibitor (G-15), eNOS inhibitor (L-NAME), and sGC inhibitor (ODQ). This study integrates pharmacology and experimental evaluation to reveal that TMYX activates HIF-1α/eNOS signaling pathway by upregulating GPER to relax coronary microvessels, thereby significantly alleviating NR.

dialysis.

Objective:To study the cognitive function and brain white matter fiber change in major depressive patients prior and post-treatment.Methods:Eleven major depressed patients were given antidepressants for 10 weeks, and their conditions were evaluated using 24-item Hamilton Depression Scale(HAMD).The cognitive function was determined by using Wisconsin Card Sorting Test(WCST),part of Wechsler memory scale and diffusion tensor ima- ging(DTI)was scanned before and after treatment.11 healthy people as control group were involved and given the same tests at the same time.Results:(1)The WCST scores of patients increased significantly after treatment(prior treatment Cc:1.6?1.6,Re:67.9?20.0,Rpe:51.5?24.8;post treatment Ce:4.0?2.1,Re:43.2?18.8,Rpe:22.8?16.0,P=0.001/0.000/0.003).There was no difference in number sequence memory in Wechsler memory scale.No difference was found between patients after treatment and control group in either WCST or number sequence memory.The patients made significant improvement in the total score of HAMD after treatment(16?14/54?13,t=6.60,P

The nucleic acid fragment of porcine circovirus type 2(PCV2)was amplified using PCR from the tissues of a domesticated wild boar with suspected postweaning multisystemic wasting syndrome(PMWS).The sample were then inoculated into the Dulac cells and passaged for 8 generation.The 4 nucleic acid fragments covered the complete genome for PCV2 were obtained by over-lapping PCR and sent to sequence.The sequence of genome was compared with other 9 strains originated from piglets in the same area.The result showed that these strains were in high homology.

Objective To discuss the effects of extract of Ginkgo BilobaLeaves(EGb) on expression of cytokine of renal interstitial fibrosis induced by transforming growth factor-?1 (TGF-?1) and extracellular matrix. Methods Cultured human kidney cells(HKC) were divided into three groups: control group,TGF-?1(8 ng/mL) group,and TGF-?1(8 ng/mL) added EGb(25,50,100,150 mg/L)group.After 72 h,expression of ?-SMA was detected by cell immunochemistry ABC,and collagen type I by Real-time PCR and Western blotting. Results Treated with TGF-?1(8 ng/mL) for 72 h,expression of ?-SMA and collagen type Ⅰ were up-regulated markedly compared with control group.Treated with EGb(25,50,100,150 mg/L)and TGF-?1(8 ng/mL)concomitantly for 72 h,expression of ?-SMA and collagen typeⅠ were down-regulated in dosage dependent manner compared with TGF-?1 group. Conclusion EGb can inhibit expression of ?-SMA and collagen type I in HKC induced by TGF-?1,and the possible mechanism might be related to the inhibition of EGb on renal tubular epithelial-myofibroblast transdifferentiation.

Background When limbal stem cell deficiency (LSCD) occurs,not only the limbal stem cells (LSCs) were damaged,but also the LSCs matrix microenvironment was under destruction.The treatment of LSCD include both replenishing of stem cells and restoration of microenvironment.So far,the method to improve the microenvironment of LSCD still exist limitation and urgently need to establish more appropriate microenvironment for the LSCs growth in vitro.Objective This study was to investigate whether the human bone marrow-derived mesenchymal stem cells (BMSCs) can be used as the ideal cells to repair limbal microenvironment and its possible mechanism of repairing limbal microenvironment during the human LSCs amplification in vitro as feeder cells.Methods BMSCs were cultured and passaged in vitro,and flow cytometry was used to assay the expressions of CD45,CD71,CD90,CD105 and HLA-DR and directionally induced BMSCs to the osteoblasts and adipocytes.BMSCs were treated using mitomycin C (MMC) to use as the feeder cells.LSCs were separately co-cultured with BMSCs,Swiss-3T3 feeder cells and free-feeder cells,and colony-forming efficiency (CFE) of the LSCs was compared among different co-cultured groups.LSCs were then cultured sequentially and identified by flow cytometry.Expression of cytokines in BMSCs was confirmed by reverse transcriptional polymerase chain reaction (RT-PCR).Results Cultured BMSCs showed a good homogeneity,with a lot of expressions of interstitial cell markers such as CD71,CD90,CD105 and less expressions of hematopoietic cell markers including CD45 and HLA-DR.After separately cocultured with feeder cells for 12 days,the CFE of the LSCs co-cultured with BMSCs,Swiss-3T3 and no feeder cells was 3.67％ ±0.58％,4.30％ ± 1.54％ and 0.20％ ±0.10％,showing a statistical significant difference among the three groups(F =15.420,P =0.040).There was no statistically significant difference in the C FE of the LSCs between the BMSCs feeder group and the Swiss-3T3 feeder cells group(P =0.456),between the BMSCs feeder group and the free-feeder cells group or the Swiss-3T3 co-culture group and the free-feeder cells group (P =0.005,0.002).LSCs presented with a positive response for ABCG2 antigen in the co-cultured with BMSCs group.Basic fibroblast growth factor(bFGF),stem cell factor (SCF) and N-cadherin(N-cad) were positively expressed in the BMSCs as feeder cells.Conclusions Human BMSCs-derived feeder cells can improve the growth of the stromal microenvironment of the LSCs and enhance their proliferation ability.Human BMSCs are suitable for engineering of epithelial sheets.

Objective To explore the clinical and pathological feature of pediatric chronic gastritis and whether gastritics in children related to hericobacter pylori(Hp).Methods Retrospective analysis the result of 401 children with gastroendosopy and 100 species with Hp detected were performed.Results Superficial gastritis form mild to moderate was the mainly pathologic change in pediatric chronic gastritis,chronic inflammation was the mainly change in pediatric Hp infection,and the detected rate of chronic superficial gastritis from mo-(derate) to heavy was significantly higher than mild(?~2=7.61 P

Objective To investigate the effects of subretinal fluid drainage combined with intravitreal anti-vascular endothelial growth factor (VEGF) drugs in the treatment of severe exudative retinal detachment Coats disease.Methods Thirteen patients (13 eyes) with 3B Coats' disease diagnosed at the Eye Center of Tongren Hospital were included in the study.The participants were aged from 1 year to 11 years with a mean age of (4.15 ± 2.99) years.The visual acuity was no light perception in 1 case,from light perception to counting finger in 7 cases,from 0.01 to 0.1 in 2 cases,and could not be measured due to young in 3 cases.Patients underwent retinal fluid drainage combined with intravitreal ranibizumab (IVR,0.5 mg,0.05 mL) at the pars plana of ciliary body,and with retinal laser photocoagulation or cryotherapy according to the retinal peripheral vascular activity.During the follow-up,the visual acuity,intraocular pressure,slit lamp,indirect ophthalmoscope and color ophthalmoscope were examined and observed.The abnormal blood vessel change,absorption of subretinal fluid,retinal reattachment and complication were observed.Results Two subretinal fluid drainage were performed in 3 cases,one subretinal fluid drainage in 10 cases.Six cases were combined with two intravitreal injections,4 cases with three intravitral injection,3 cases with intravitreal injection for more than three times.Five cases were treated with simple photocoagulation,3 cases with simple retinal cryotherapy,and 5 cases with laser combined with cryotherapy.In 13 patients,the visual acuity was improved in 2 cases,unchanged in 8 cases,and could not be measured due to young in 3 cases.Eight cases had complete retinal reattachment.No significant postoperative complications occurred during follow-up,such as endophthalmitis,retinal hole and vitreous hemorrhage.Conclusion Subretinal fluid drainage combined with intravitreal injection is an effective method for severe 3B stage Coats disease.

Objective To establish a tree shrew model of Fusarium solani keractitis by injecting Fusarium solani conidia into the corneal stroma.Methods Fusarium solani was inoculated into Sabouraud culture medium and incubated at 26℃ for 7 days.Fungal suspension was collected and the number of spores was adjusted to 1 × 1010 CFU /mL on the blood cell count plate.Forty healthy tree shrews were randomly divided into experimental group (n=30) and control group (n=10).In the experimental group, 50 μL of fungal spore suspension was injected into the cornea center with a 29G needle, and 50 μL saline was injected in the control group.The models were evaluated by anterior segment photography, in vivo confocal microscopy, histopathology, and corneal tissue culture.Results The fungal infiltration, the degree of edema of corneal epithelial and endothelial cells, and the number of mycelium were positively correlated with time.The number of infiltrating inflammatory cells, mainly, neutrophils, reached a peak on the 7th day after modeling.The mycelial growth was parallel to the stromal fibers.After the successful establishment of the model, the corneal tissue culture showed the growth of Fusarium solani.The successful rate of modeling was 86%.Conclusions The tree shrew model of Fusarium solani keratitis is established by injecting spores of Fusarium solani into the cornea.