To investigate the effects of Pithecellobium clypearia extract on the tissue structure,in-flammatory lesions as well as microbial diversity in the intestinal of yellow-feathered broilers.2401-day-old yellow-feathered broilers were randomly divided into four groups(groups A,B,C and D),groups A,B and C were supplemented with Pithecellobium clypearia extract in basal diets with concentrations of 0.5,1.0 and 2.0 g/kg,respectively.Group D served as the control group without adding Pithecellobium clypearia extract in diets,and the full trial period lasted for 70 d.Duodenum and jejunum samples were collected on the 20th,40th and 70th days of the test,the vil-lous/crypt ratio of duodenum and jejunum were calculated,and the mRNA expression level of in-flammatory cytokine as well as related pathways were detected in each group,respectively.In addition,the contents of cecum were collected at 70 th day of the experiment and the microbial di-versity in cecum were also analysed by 16S rDNA sequencing.The results showed that adding 0.5 and 1.0 g/kg of Pithecellobium clypearia extract in the diet could significantly increase the veloci-ty height/crypt depth ratio of duodenum and jejunum(P＜0.05)compared to control group,as well as the mRNA expression level of tight junction protein(CLDN1 and CLDN5)in jejunum,which further improved the structure of mucous of intestinal.Pithecellobium clypearia extract could significantly(P＜0.05)decrease the mRNA expression level of inflammatory cytokine inclu-ding IL-1β,IL-8 and TNF-α,as well as the related pathway genes such as TLR4,MyD88 and NF-κB in jejunum,thus reduced the inflammatory lesions in intestinal.Pithecellobium clypearia ex-tract also could significantly increase the abundance of beneficial microbial such as Parabacteroide and Prevotellaceae,while significantly decrease the abundance of pathogenic microbial such as Proteobacteria in cecum(P＜0.05)and improve the microbial diversity in intestinal.In summary,Pithecellobium clypearia extract could improve the structure of intestinal tissue and the gut barri-er function,as well as the microbial diversity in cecum,and also decrease the inflammatory lesions in jejunum,which is helpful to the intestinal health for yellow-feathered broilers.The present study provides scientific basis for the development of Pithecellobium clypearia as a safe feed additive in the future.

The standardized workflow of computer-aided static guided implant surgery includes preoperative exami-nation,data acquisition,guide design,guide fabrication and surgery.Errors may occur at each step,leading to irrevers-ible cumulative effects and thus impacting the accuracy of implant placement.However,clinicians tend to focus on fac-tors causing errors in surgical operations,ignoring the possibility of irreversible errors in nonstandard guided surgery.Based on the clinical practice of domestic experts and research progress at home and abroad,this paper summarizes the sources of errors in guided implant surgery from the perspectives of preoperative inspection,data collection,guide de-signing and manufacturing and describes strategies to resolve errors so as to gain expert consensus.Consensus recom-mendation:1.Preoperative considerations:the appropriate implant guide type should be selected according to the pa-tient's oral condition before surgery,and a retaining screw-assisted support guide should be selected if necessary.2.Da-ta acquisition should be standardized as much as possible,including beam CT and extraoral scanning.CBCT performed with the patient's head fixed and with a small field of view is recommended.For patients with metal prostheses inside the mouth,a registration marker guide should be used,and the ambient temperature and light of the external oral scan-ner should be reasonably controlled.3.Optimization of computer-aided design:it is recommended to select a handle-guided planting system and a closed metal sleeve and to register images by overlapping markers.Properly designing the retaining screws,extending the support structure of the guide plate and increasing the length of the guide section are methods to feasibly reduce the incidence of surgical errors.4.Improving computer-aided production:it is also crucial to set the best printing parameters according to different printing technologies and to choose the most appropriate postpro-cessing procedures.

Objective To investigate the effects of homocysteine(Hcy) on myocardial injury and its possible mechanisms.Methods The selected H9C2 cardiomyocytes were intervened with various concentrations of Hcy and 4-phenyl butyric acid(4-PBA).The H9C2 cells were divided into the control group,H400 group and H400P2 group.The control group used the common medium,the H400 group was added with 400 μmol/L Hcy,the H400P2 group was added with 2 mmol/L 4-PBBA on the basis of H400 group.The cell livability was detected by using cell counting kit-8 (CCK-8).Apoptosis was evaluated by using the terminal deoxynucleotidyl transferase mediated nick-end labelling(TUNEL) staining.The ERO1α expression was determined by using immunocytochemistry,and the protein expression difference was determined by using Western blot.Results The injury of Hey on H9C2 cardiomyocytes showed a concentration-dependent manner(F=2 039.958,P＜0.01).Compared with the control group,the apoptosis percentages and expression levels of PERK,p-PERK,CHOP and ERO1α in the H400 group were increased(P＜0.01);while which in the H400P2 group were decreased,the difference was statistically significant(P＜0.05).Conclusion Hcy mediates myocardial apoptosis through endoplasmic reticulum stress mechanism.

Objective To observe the influence of different level of hyperhomocysteinemia on mRNA and protein expressions of KV1 .3 ,CaN,NFAT,IL-6 and TNF-αin lymphocytes of patients with acute ST-segment elevation myocardial infarction (STEMI).Methods We selected 90 STEMI patients and divided them into three groups according to the level of plasma homocysteine:the first experimental group (STEMI group,Hcy<1 5μmol/L, n=30),the second experimental group (STEMI with mild Hhcy group,Hcy 15~30μmol/L,n=30)and the third experimental group (STEMI with intermediate Hhcy group,Hcy>30 μmol/L,n=30 ).Another 30 healthy examined people were selected as control group (n=3 0 ).Peripheral lymphocytes were isolated by Ficoll density gradient centrifugation.The Hcy in the plasma was measured with the IMX assays.Real-time quantitative PCR (RT-PCR)was used to detect mRNA expressions of KV1.3,CnAα,NFAT1,IL-6 and TNF-αand Western blot technique was used to detect the expressions of KV1.3,CnAαand NFAT1.Results The mRNA and protein expression levels of KV1.3,CnAαand NFAT1 in each experimental group were significantly higher than those in control group (P<0 .0 5 or P<0 .0 1 ).Multiple comparison in each experimental group showed that compared with that in the first experimental group,the expression level of the second experimental group increased (P<0.05 or P<0.01)and compared with first and second experimental groups,the expression level of the third experimental group increased (P<0.05 or P<0.01).The mRNA expression levels of IL-6 and TNF-αin each experimental group were significantly higher than those in control group (P<0.05 or P<0.01 ).Multiple comparison in each experimental group showed that compared with that in the first experimental group,the expression level of the second experimental group increased (P<0 .0 5 or P<0 .0 1 )and compared with first and second experimental groups,the expression level of the third experimental group increased (P<0.01).Plasma total Hcy levels were positively correlated with mRNA and protein expressions of KV1.3 in all observed groups (r=0.503 P=0.000,r=0.726 P=0.000).Conclusion The higher level of Hcy in plasma,the higher mRNA and protein expression levels of KV1.3,CnAα,NFAT1 and the higher mRNA expression levels of IL-6,TNF-αin the lymphocyte of STEMI patients,which may be one mechanism for Hcy exacerbating the inflammatory reaction of STEMI.

[ ABSTRACT] AIM:To investigate the effects of Chaihu Shugan powder ( CSP) on lipid metabolism and the pro-teins involved in adenosine 5’-monophosphate-activated protein kinase (AMPK)/sirtuin 1 (SIRT1) pathway in the liver tissues of the rats with non-alcoholic fatty liver disease (NAFLD).METHODS: Sprague-Dawley rats were randomly di-vided into normal control ( NC) group, with HFD ( HFD) group and CSP group.The NAFLD models were established by feeding with HFD for 16 weeks in the rats.The rats in CSP group were intragastrically administered with CSP extracts (9.6 g· kg-1 · d-1 ) , and blood and liver samples were collected 16 weeks later.Serum and liver levels of total cholesterol ( TC) and triglyceride ( TG) , and serum levels of alanine aminotransferase ( ALT) and aspartate aminotransferase ( AST) were measured using an automatic biochemical analyzer.The histological changes of liver tissues were observed with HE staining, while the lipid deposition was observed with Oil Red O staining.The ultrastructural changes of the liver tissues were observed under transmission electron microscope.Moreover, the protein levels of AMPK, phosphorylated AMPK (pAMPK), SIRT1 and uncoupling protein 2 (UCP2) in the liver were detected by Western blot.RESULTS:The results of HE staining, Oil Red O staining and electron microscopy demonstrated that NAFLD rat model was successfully estab-lished.Compared with NC group, the serum and liver levels of TC and TG, and serum level of AST in model group were markedly elevated ( P<0.01) .Moreover, the protein levels of pAMPK and SIRT1 in HFD group were markedly reduced (P<0.01), whereas UCP2 level was elevated (P<0.01).Furthermore, liver levels of TC and TG, and serum level of AST in GSP group were markedly reduced as compared with HFD group ( P<0.05 ) .The protein levels of pAMPK and SIRT1 were elevated ( P<0.05 ) , whereas the UCP2 level was reduced as compared with HFD group ( P<0.01 ) .The protein level of AMPK between the 3 groups had no significant difference.CONCLUSION: CSP attenuates hepatic lipid disorder and hepatic lipid deposition in NAFLD rats induced by feeding with HFD for 16 weeks, which is associated with the activation of AMPK/SIRT1 pathway.

Objective To investigate the relationship between plasma homocysteine (Hcy), Kv1.3 channel and cardiac troponinI (cTnI) in patients with acute ST-segment elevation myocardial infarction (STEMI). Methods According to the level of Hcy, 80 STEMI patients were divided into STEMI with Hhcy group (Hcy > 15 μmol/L, n=41) and control group (STEMI group, Hcy≤15μmol/L, n=39). The Hcy, blood lipid and cTnI were detected with automatic biochemistry analyzer, respectively. Peripheral lymphocytes were isolated by ficoll density gradient centrifugation. Real-time PCR was used to detect mRNA expression of Kv1.3, and Western blot assay was used to detect protein expression of Kv1.3. Results cTnI concentrations were obviously higher in STEMI with Hhcy group than those in STEMI group (μg/L:22.997 ± 5.880 vs. 12.881 ± 6.343;P<0.01). Multiple linear regression analysis showed that age, gender, hypertension, diabetes mellitus, smoking, family history, total cholesterol (TC), triglyceride (TG), high density lipoprotein-cholesterol (HDL-C) and low density lipoprotein-cholesterol (LDL-C) had no obvious influence on Hcy (P>0.05). The relative expression levels of Kv1.3 mRNA and protein were significantly higher in STEMI with Hhcy group (1.35±0.14, 0.85±0.12) than those in STEMI group (1.00 ± 0.07, 0.64 ± 0.05, P<0.05). Moreover, there was a positive relation between Hcy level and the mRNA and proteinexpression of Kv1.3 channel (r=0.299, r=0.542, P<0.05). There was a positive relation between protein expression levels of Kv1.3 channel and cTnI (r=0.644, P<0.05). Conclusion Our results support that Hcy could exacerbate the concentration of cTnI through playing an important role in the Kv1.3 mRNA and protein expression in lymphocytes.

The purpose was to optimize the vitrification for porcine embryos cryopreservation. Blastocyst/Morula (5-6th day-embryos) were collected from superovulated Bama mini-pigs (sows/gilts). We compared different cryopreservation methods, cryopreservation tools, thining of zona pellucida (ZP) and recipient breeds on the efficiency of porcine embryo cryopreservation. The results showed that: in embryo survival rate and blastocyst cell number, there were no significant differences between cryopreservation method I [embryos were vitrified by two step method with open pulled straw (OPS) and glass micropipette (GMP) in solution 1 (TCM199 + 20% FBS + 10% EG + 10% DMSO) for 3 min, and solution 2 (TCM199 + 20% FBS + 20% EG + 20% DMSO + 0.4 mol/L SUC) for 1 min, stored in liquid nitrogen] and method II[Blastocysts were cultured for 25 min in NCSU23 + 7.5 microg/mL cytochalasin B, centrifuged at approximately 13 000 xg for 12-13 min, and recovered back into pNCSU23. They were then equilibrated for 5 min in 2 mol/L ethylene glycol in pNCSU23, washed quickly in the vitrification medium, 8 mol/L ethylene glycol, 7% polyvinylpyrrolidone (PVP) in pNCSU23, loaded into OPS/GMP, and plunged into liquid nitrogen]. GMP vitrification method was more suitable and efficient than OPS method (P < 0.05) in embryo survival rate (83.8% vs 77.6%) and blastocyst cell number (53.1 vs 47.5) after thawing. Thining of ZP did not increase the survival rate, but significantly improved blastocyst cell number in the survival blastcysts (60.1 and 46, P < 0.01). Local pig breeds (Fengjing sows) were more suitable as recipients for embryo transfer of vitrified/warmed blastcysts, which can improve pregnant rate and embryo efficiency.
Animals ; Blastomeres ; cytology ; Cryopreservation ; methods ; veterinary ; Embryo Transfer ; veterinary ; Embryo, Mammalian ; Swine ; Swine, Miniature ; Vitrification ; Zona Pellucida ; physiology

Objective To investigate analgesic effects of the selective blocking of descending facilitation targeting μ opioid receptor positive neurons in the rostral ventromedial medulla ( RVM) in a rat model of bone cancer pain. Methods Forty-eight adult female Wistar rats weighing 180-200 g were randomly divided into 6 groups: group Ⅰ control ( n = 3) ;group Ⅱ bone cancer pain induced by intra-tibia inoculation of Walker 256 mammary gland carcinoma cells ( n = 9) ;group Ⅲ-Ⅵ received a single intra-RVM micro-injection of PBS (group Ⅲ), dermorphin (group Ⅳ) , saporin (group Ⅴ) and dermorphin-saporin ( group Ⅵ) respectively at 28 days before intra-tibia inoculation ( n = 9 each) . Starting from 3 to 20 days after intra-tibia inoculation, mechanical allodynia was assessed and recorded. The animals were sacrificed on 7, 14 and 20 days after intra-tibia inoculation, after repetitive non-noxious tactile stimulation of the hindpaw. The total number of Fos-positive neurons in the spinal dorsal horn was measured as a marker indicative of central sensitization. Results The animals developed nociceptive hypersensitivity after intra-tibia cancer cell inoculation in group Ⅱ -Ⅵ . Nociceptive hypersensitivity was significantly decreased during 4-7 days after the onset of nociception in group Ⅵ (dermorphin-saporin). The number of Fos positive neurons in bilateral spinal dorsal horn was significantly increased by intra-tibia inoculation of cancer cells in group Ⅱ-Ⅵ as compared with control group and was significantly lower at day 14 and 20 after inoculation in group Ⅵ (dermorphin-saporin) than in group Ⅱ - Ⅴ.Conclusion Selective blocking of descending facilitation targeting μ opioid receptor positive neurons in RVM can effectively reduce nociceptive hypersensitivity induced by intra-tibia inoculation of Walker56 mammary gland carcinoma cells.

This is a study on the biodegradable polymers as gene controlled-released coatings for gene transfer. The PELA (poly (Dl-lactic acid)-co-poly (ethylene glycol), and PLGAE (poly (lactic acid)-co-poly (ethylene glycol)-co-poly (glycolic acid) random copolymer) were synthesized and prepared as the coatings of plasmid pCH110 in the transfection. All kinds of factors affecting the loading efficiency, cytotoxicity, transfection efficiency and the course of the degradation and release in vitro were discussed. The average diameters of microspheres of PELA and PLGAE were 1-3 microm and 0.72 microm respectively. The loading efficiency levels of them were 62% and 70% respectively. The transfection efficiency levels of two kinds of pCH110 delivery system for COS-1 cells were higher and two of them had few cytotoxicity. After transfection, the X-gal assay was performed and reported positive for 96 h. The biodegradable polymeric materials as gene carriers possess their potential superiority.
Biocompatible Materials ; DNA ; chemistry ; Drug Carriers ; chemical synthesis ; chemistry ; toxicity ; Gene Transfer Techniques ; Lactates ; chemical synthesis ; chemistry ; toxicity ; Lactic Acid ; chemical synthesis ; chemistry ; toxicity ; Polyethylene Glycols ; chemical synthesis ; chemistry ; toxicity ; Polyglycolic Acid ; chemical synthesis ; chemistry ; toxicity ; Polymers ; chemical synthesis ; chemistry ; toxicity ; Transfection

Salmonella Typhi capsular polysaccharide vaccines were encapsulated in the Micro-particles made from polyethylene glycol-poly-DL-lactide (PELA). BALB/c mouse were divided into three groups with 20 mice in each. Mouse were immunized respectively with controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines and Salmonella Typhi capsular polysaccharide vaccines by oral and subcutaneous administration. The mice blood and salvia were collected at the 2nd, 4th and 8th weeks respectively for the titrating of IgG and sIgA antibodies by RIA. At the 8th week, live typhoid bacteria were injected into the immunized mice for the calculation of the rate of immunization protection. The IgG titers of the controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines group were higher than those of the other groups(P < 0.05). The IgA titers of the low groups of controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines (oral and subcutaneous) were higher than those of the group of Salmonella Typhi capsular polysaccharide vaccines (P < 0.05). The immunization protection rates of the three groups were 40%, 100% and 60% respectively. The controlled release microencapsulated Salmonella Typhi capsular polysaccharide vaccines possess the advantages of releasing slowly in vivo and persisting long time immunogenicity.
Administration, Oral ; Animals ; Delayed-Action Preparations ; Female ; Immunoglobulin A, Secretory ; analysis ; Immunoglobulin G ; blood ; Injections, Subcutaneous ; Mice ; Mice, Inbred BALB C ; Microspheres ; Polysaccharides, Bacterial ; administration & dosage ; immunology ; Typhoid-Paratyphoid Vaccines ; administration & dosage ; immunology ; Vaccination