Mannheimia haemolytica is an important pathogen causing respiratory diseases in sheep,and the development of an inactivated vaccine of sheep origin against Mannheimia haemolytica is an effective means of prevention and control of this pathogen.In this study,Mannheimia haemo-lytica isolated from the diseased material collected from a sheep farm was screened for the most suitable vaccine strain by mouse pathogenicity test,and the inactivated vaccine was prepared by combining four antigenic amounts(106,107,108,109 CFU/mL)with three adjuvants(ISA 201 ad-juvant,ISA 206 adjuvant,and aluminum adjuvant),which were used in immunization and attack protection tests,and then screened for immunity and attack protection tests.Immunization and at-tack protection tests were conducted to screen the optimal antigen amount and adjuvants for inacti-vated vaccine preparation.Finally,the vaccine was prepared under the optimal conditions and the dynamic changes of antibody levels in immunized sheep were observed.The results showed that a total of 24 strains of Mannheimia haemolytica of sheep origin were isolated,among which 12 strains of serotype A1 were the dominant serotypes for the infection in this sheep farm;one strain of Mannheimia haemolytica of sheep origin of type A1(Mh1)was screened and obtained as the strain for vaccine preparation.Its LD50 for mice was 3.16 × 107 CFU/mL,with strong pathogenici-ty,and could cause pathological damage such as inflammatory cell infiltration of varying degrees in heart,liver,spleen,lung and kidney tissues;the results of the immunization and attack protection test showed that the optimal antigenic amount of the inactivated vaccine was 109CFU/mL,and the most suitable adjuvant was ISA 201,which stimulated the production of the highest antibody level and provided 100％protection for the mice.The protection rate of mice reached 100％,and good humoral immunity could be induced after immunizing sheep.The inactivated Mannheimia haemo-lytica type A1 vaccine prepared in this study has good immunoprotection effect,and lays a good foundation for the further development of commercial vaccine against Mannheimia haemolytica of sheep origin.

Background: Enteritis is one of the most frequently reported symptoms in piglets infected with porcine circovirus type 2 (PCV2), but the immunopathogenesis has not been reported. Objectives: This study examined the effect of a PCV2 infection on the intestinal mucosal immune function through morphological observations and immune-related molecular detection. Methods: Morphological changes within the ileum of piglets during a PCV2 infection were observed. The expression of the related-molecules was analyzed using a gene chip. The immunocyte subsets were analyzed by flow cytometry. The secretory immunoglobulin A (SIgA) content was analyzed by enzyme-linked immunosorbent assay. Results: The PCV2 infection caused ileal villus damage, intestinal epithelial cells exfoliation, and an increase in lymphocytes in the lamina propria at 21 days post-infection.Differentially expressed genes occurred in the defense response, inflammatory response, and the complement and coagulation cascade reactions. Most of them were downregulated significantly at the induction site and upregulated at the effector site. The genes associated with SIgA production were downregulated significantly at the induction site. In contrast, the expression of the Toll-like receptor-related genes was upregulated significantly at the effector site. The frequencies of dendritic cells, B cells, and CD8 + T cells were upregulated at the 2 sites. The SIgA content decreased significantly in the ileal mucosa. Conclusions PCV2 infections can cause damage to the ileum that is associated with changes in immune-related gene expression, immune-related cell subsets, and SIgA production.These findings elucidated the molecular changes in the ileum after a PCV2 infection from the perspective of intestinal mucosal immunity, which provides insights into a further study for PCV2-induced enteritis.

Objective? To? explore? the? clinical? significance? of? early? oral? intervention? measures? in? the?prognosis?of?premature?infants.? Methods? 151?preterm?infants?admitted?to?neonatal?intensive?care?unit?(NICU)?of?Liaocheng?People's?Hospital?from?January?2015?to?January?2017?were?enrolled.?Premature?infants?were?divided?into?intervention?group?and?control?group?according?to?random?number?table?method?and?with?the?consent?of?legal?guardian.?Both?groups?received?routine?treatment?of?preterm?infants?after?stable?vital?signs.?The?intervention?group?received?the?oral?massage?method?adopted?by?none-nutritive?sucking,?stimulating?swallowing?function?and?SandraFucile?on?the?basis?of?routine?treatment,?once?a?day?for?14?consecutive?days.?Both?groups?were?followed?up?for?6?months.?The?oral?feeding?ability?of?premature?infants?was?evaluated?by?the?proficiency?(PRO),?rate?of?transfer?(RT),?feeding?process?and??non-nutritive?suction?(NNS).?At?40?weeks?of?postmenstrual?age?(PMA),?neonatal?behavioral?neurological?(NBNA)?was?used?to?assess?neonatal?brain?development;?Infanib?was?used?for?early?motor?development?evaluation?at?3?months?and??6?months?after?birth.? Results? Finally,?151?premature?infants?were?enrolled,?including?78?in?the?intervention?group?and?73?in?the?control?group.?The?time?to?complete?oral?feeding?of?the?intervention?group?was?significantly?shorter?than?that?of?the?control?group?(days:?18.1±3.7?vs.?23.4±5.8,?P??0.05).?Infanib?evaluation?at?3?months?of?age?showed?that?the?proportion?of?normal?children?in?the?intervention?group?was?significantly?higher?than?that?in?the?control?group?[67.95%?(53/78)?vs.?49.31%?(36/73),?P?

Objective To investigate the effect of triptolide (TP) on proliferation and apoptosis of cyst-lining epithelial cells with autosomal dominant polycystic kidney disease ( ADPKD). Methods Primary cultured cyst-lining epithelial cells were treated with TP at different concentrations for 12 h,24 h,48 h and 72 h, respectively.The proliferation activity of the cells was evaluated by Brdu assay. The cell cycle distribution was determined by flow cytometry. The apoptotic and apoptotic ratio were determined by FITC-AnnexinV binding/ PI. The morphological changes of cyst-lining epithelial cells were observed under transmission electron microscope. Results TP significantly inhibited the proliferation of cyst-lining epithelial cells and induced apoptosis in a dose- (10-40 ng?mL-1 )and time-dependent(12-48 h) manner. Typical ultrastructural changes of apoptotic cells were observed under electron microscope. Conclusion TP significantly inhibited the proliferation of cyst-lining epithelial cells and induced the apoptosis of cyst-lining epithelial cells, thus inhibited cyst forming and delayed cyst developing. The mechanism may involve several targets and pathways.

BACKGROUND:The umbilical cord blood is rich in hematopoietic stem/progenitor cel s that have strong proliferation and differentiation ability as wel as ability to form colonies, and exert important roles in stimulating bone marrow function, improving blood cel viability and quantity, promoting immune cel maturation, and maintaining immune balance.
OBJECTIVE:To evaluate the clinical effects of autologous umbilical cord blood mononuclear cel transplantation on the immunologic function and prognosis for premature infants.
METHODS:Sixty-two preterm infants who entered into NICU immediately after birth, weighing ≤ 1 500 g, were divided into treatment group and control group according to parent’s wil ingness. In the treatment group, the umbilical cord blood was extracted from the umbilical vein and re-infused into the preterm infants after density gradient centrifugation within 4 hours. The cel ular immunity levels, humoral immunity levels and clinical parameters were monitored before and after treatment.
RESULTS AND CONCLUSION:After 1 week of treatment, the CD4, CD4/CD8 levels were significantly increased compared with the control group (P=0.01, 0.03), but CD8 level had no changes. At 1 week after treatment, IgM levels were both increased in the two groups, especial y in the control group (P=0.00);IgA levels had no changes;IgG levels were decreased, especial y in the control group (P=0.02). The incidence of severe infection during hospitalization was 13%in the treatment group, which was lower than the control group (16%), but there was no difference between the two groups. The proportion of infants undergoing mechanical ventilation and average length of stay had significant differences between the two groups (P<0.05). After 12 months, the incidence of recurrent respiratory tract infections was zero in the treatment group and one case in the control group, and there was a significant difference between the two groups. These findings indicate that autologous umbilical cord blood mononuclear cel transplantation can improve the immunologic function, slower the reduction of IgG levels, reduce the usage of breathing machine, shorten the length of stay, and reduce the incidence of recurrent respiratory tract infections in preterm infants.

Objective To investigate the clinical features,treatment and the causes of death in severe infectious disease complicated with capillary leak syndrome(CLS)in neonates.Methods The clinical data,laboratory finding,treatmand clinical outcome of 11 neonates who had severe infectious disease complicated with CLS in our NICU from Jan 2009 to Jul 2010 were collected and analysed retrospectively.Results Among the 11neonates,five had pneumonia and the other six had sepsis.All the 11 cases appeared progressive edema on skin and mucosa,dyspnoea,infective shock,oliguria and hypoalbuminemia(10～20 g/L).We treated the 11cases with hydroxyethyl starch(10～15 ml/kg,every 8 to 12 h)at early stage on the basis of infection control,anti-shock treatment,mechanical ventilation,symptomatic treatment and a stable internal environment.At last,6 cases were rescued,in whom 4 cases were well-developed,had normal intelligence and athletic ability,the other two cases had hydrocephalus or muscle tone high of both lower extremities.Five cases died.Conelusion CLS is a severe complication of neonatal severe infection,and had high mortality.Treating with hydroxvethvl starch at an early stage can increase the survival rate.

Objective To invest the effect of minimal inflammation on patients with maintenance hemodialysis(MHD) and the intervention effect of ftuvastatin.Methods Blood indicators such as PA,ALB and BUN,Cr,TC,TG,HDL-C,LDL-C,Lp(a),SF,hs-CRP,IL-6 and TNF-α were detected at the first day of the study and six months later in all patients.Patients were divided into inflammation group(CRP≥8 mg/L) and non-inflammation group(CRP<8 mg/L) according to CRP levels.Only inflammation group used fluvastatin.Two groups were observed for six months.Results The level of hs-CRP,IL-6,TNF-α,TG,Lp(a),SF in MHD patients was obviously higher than the normal control group.The level of ALB was obviously lower than the normal control group.The angiocardiopathy incidence of inflammation group was obviously higher than that of the non-inflammation.The more obvious of minimal inflammation,the more significant of lipid metabolism disturbance and malnutrition.Hs-CRP,IL-6,TG,TC,LDL-C and Lp(a) of inflammation group decreased obviously(P<0.01 or P<0.05).ALB、HDL-C increased obviously(P<0.05).Indicators of the non-inflammation had no statistical difference after therapy.Conclusions Patients with MHD generally have minimal inflammation and malnutrition.Minimal inflammation has important effect on the angiocardiopathyincidence,lipid metabolism disturbance and malnutrition.Fluvastatin not only could regulate lipid metabolism but also improve the minimal inflammation of patients with MHD.Early detection and therapy of minimal inflammation has important significance on improving prognosis of patients with MHD.

Objective To ivevstigate the feasibility, the effects and the therapy of peritoneal dialysis(PD) on severe acute pancreatitis (SAP). Method From January,2001 to Jaly 2006,48 patients of SAP were divided randomly into PD group and non-PD group in Qingzhou hospital. Both groups were treated by the conventional mode of therapy. In PD group , using the concept of PD,24 patients of SAP were treated with PD and NPD group were treated only with common therapy. The release time of abdominal pain and distention, CT scores, APACHE II scores, the time of hospital stay, cost of treatment in hospital, operative rate and rate of complications and recovered rate of the two groups were compared. Simutaneously. the concentration of serum and fluid filtrated inflammatory cytokines TNFα,IL-6,IL-8,IL-10 and CRP were also determined pro and post the therapy. Results In the PD and NPD group, the duration for disappear-ance of abdominal pain and tenderness,and amelioration for abdominal distension was (19.70 ±7.32) hvs. (81.46±36.68) h and (23.16±6.95) h vs.(78.19± 29.26) h;So that the PD group was precede to that in NPD group ( P < 0.05). The concentration of serum and fluid filtrated pro-inflammatory cytokines TNFα, IL-6, IL-8 and CRP at each observation points after PD was decreased significantly (P <0.05) in the PD group. But the concentration of the serum and fluid filtrated anti-inflammatory cytokines IL-10 was increased significantly (P < 0.05) as compared with that of the NPD group. Conclusions Through PD, the imbalance of pro-inflammatory and anti-inflammatory cytokines has been corrected at early stage of SAP. But the PD is method that is easy, quick, small wou-ndell,few complicated and effective in patients with early-phase SAP, and will be of great value in wide application and further study.

Objective To investigate the significance of renal tubular dysfunction in patients with refractoriness nephrotic syndrome(RNS)and the effect of interference treatment of Valsartan (VAL).Methods 79 cases of RNS and 68 healthy controls were recruited into the study. The 79 patients of RNS were divided randomly into the VAL group and the dipyridam group. On the basis of routine therapy, the VAL group was given VAL(80mg/d),and the dipyridam group taken dipyridam (150mg/d)orally for 12 weeks The glomerular tubular function(u-RBP,α1-MG,β2-MG,mAlb,NAG)were detected and the pathologic changes of tubukinterstitium were observed by using the methods of ELISA, biochemistry and scoring of the pathologic damage of tubulointerstitium before and after 12 weeks of VAL treatment in all of the cases. Results Urinary RBP,α1-MG,β2-MG,mAlb and NAG in all patients with RNS was evidently higher than that in healthy controls(P＜0.01).Those had positive correlations with damages of tubulointerstitium(r=0.436,0.626,0.499,0.668,0.657,P＜0.01).The interference outcome displayed that the excretion rates of urinary series of protein after oral use VAL in treatment group were markedly lower, while the control group had no distinct change. Conclusion There were various injury of tubulointerstitium and the disfiguration of renal tubular function in all cases with RNS. Damages of renal tubular function had positive correlation with tubulointemtitium injury and renal globular injury. Interference treatment with ARB in patients with RNS could improve renal tubular function, which is of great significance in delaying the progress of RNS.

In order to study the in vitro culture and expansion of bone marrow mesenchymal stem cells in rats (rMSCs) and the possibility of rMSCs differentiation into retinal neural cells, the bone marrow-derived cells in SD rats were isolated and cultured in vitro. The retinal neural cells in SD rats were cultured and the supernatants were collected to prepare conditioned medium. The cultured rMSCs were induced to differentiate by two steps. Immunofluorescence method and anti-nestin, anti-NeuN, anti-GFAP and anti-Thy1.1 antibodies were used to identify the cells derived from the rMSCs. The results showed that the in vitro cultured rMSCs grew well and expanded quickly. After induction with two conditioned media, rMSCs was induced to differentiate into neural progenitor cells, then into retinal neural-like cells which were positive for nestin, NeuN, GFAP and Thy1.1 detected by fluorescence method. The findings suggested that rMSCs could be culture and expanded in vitro, and induced to differentiate into retinal neural-like cells.
Animals, Newborn ; Bone Marrow Cells/*cytology ; Cell Differentiation/*physiology ; Cells, Cultured ; Mesenchymal Stem Cells/*cytology ; Neurons/*cytology ; Rats, Sprague-Dawley ; Retina/*cytology