Objective To observe the contribution of acute normovolemic hemodilution ( ANH) in experiment of cryopreserved dog limb replantation.Methods Sixteen healthy Beagle dogs (male:female=1:1) were divided into two groups.Dogs in the experiment group ( Group B) received ANH in the limb replantation, and dogs in the control group ( Group A) received the same amount of lactate Ringer’ s solution intravenously during the surgical operation.We recorded and compared the hemodynamic indexes, HB, HCT, the resuscitation time, the first rising head time, the first standing time and the first eating and drinking time between the two groups.Results ( 1 ) During the operation, both PaO2 and PCO2 in the two groups were normal, as well as the breathing rate.The heart rate in the group B was lower than that in the group A.Before blood transfusion, there was no statistically significant difference in HB and HCT between the two groups, but after transfusion they were significantly higher in the group B than in the group A.(2) The resuscitation time, the first rising head time, the first standing time and the first eating and drinking time of the group B were all better than those in the group A.Conclusions In cryopreserved dog limb replantation experiments, acute normovolemic hemodilution is helpful to improve the general condition and facilitate the recovery of animals after limb replantation.

Objective To investigate the regulatory effects and possible mechanisms of autophagy induction and bactericidal activity in muring bone marrow derived macrophages ( BMDMs) . Methods Murine BMDMs were isolated and cultured in vitro. Then standard strain of E. coli was used to infect BMDMs and cellular autophagy levels and AMPK activation were detected. We next modulated functions of AMPK by pretreating cells with the specific agonist or antagonist of AMPK. Then cellular AMPK activation, autophagy levels and bactericidal activity were observed after E. coli infection. Results In E. coli infected BMDMs,autophagy related molecules like LC3B and Beclin1 were upregu-lated,accompanied with elevated LC3Ⅱ/Ⅰratio. Phosphorylation of AMPK was also upregulated by E. coli treatment. Enhanced AMPK activi-ty by its agonist leads to increased cellular autophagy and bactericidal activity,whereas inhibition of AMPK by its suppressor downregulated autophagy and dampened bactericidal activity. Conclusion AMPK and its related signaling pathway is required for anti-bacterial response in macrophage,which is dependent on its function in upregulating autophagy related molecules and inducing autophagy.

Objective To investigate the differences in the biochemical and pathological features and therapeutic response between the elderly-onset autoimmune hepatitis (AIH) and young-onset AIH.Methods From October 2013 to November 2016,a total of 87 consecutive AIH patients diagnosed by liver biopsy were enrolled and divided into elderly-onset group (≥60 years) and the young-onset group (＜ 60 years).The biochemical and pathological features and therapeutic response of the two groups were compared.Mann Whitney test and chi square test were performed for statistical analysis.Results As AIH diagnosed,the proportion of liver cirrhosis detected by imaging examination of elderly-onset group and the young-onset group were 67.7％(21/31) and 35.7％(20/56),respectively,the former was higher than the latter,and the difference was statistically significant (X2 =8.214,P=0.004).The proportion of patients complicated with extrahepatic autoimmune diseases of the elderly-onset group was 9.7％ (3/31),which was lower than that of the young-onset group (28.6％,16/56),and the difference was statistically significant (X2 =4.173,P=0.041).The median levels of alanine aminotransferase (ALT) and albumin of the elderly-onset group were 50.0 U/L (22.0 U/L,193.0 U/L) and 34.8 g/L (31.3 g/L,40.5 g/L),which were lower than those of the young-onset group (146.0 U/L (43.8 U/L,390.5 U/L),39.4 g/L (35.8 g/L,44.6 g/L),and the differences were statistically significant (Z=-2.109,-2.092;both P＜ 0.05).Between the two groups,there was no statistically significant difference in inflammation grade,fibrosis stage,interface hepatitis,lymphoplasmacytic infiltration,positive rate of hepatocyte rosette pattern,positive rate of centrilobular necrosis and small bile duct injury (all P＞0.05).After treatment,among 17 patients of elderly-onset group,seven patients obtained biochemical remission and the median time to remission was 3.2 months;among 37 patients of young-onset group,18 patients (48.6％) obtained biochemical remission and the median time to remission was 2.3 months.There was no significant difference in the percentage of patients achieved biochemical remission and the median time between the two groups (both P＞0.05).Conclusion The percentage of complicated with extrahepatic autoimmune diseases of elderly-onset group is lower than that of young-onset group,which indicates that age-associated immune dysfunction may involve in the genesis and development of elderly-onset AIH.

Objective Neuroinflammation following traumatic brain injury (TBI) may give rise to neurodisorder.This study aimed to investigate the effect of intranasal delivery of nerve growth factor ( NGF) on neuroinflammation following TBI and its action mechanism in rats. Methods Thirty-six male adult Sprague-Dawley rats were equally divided into a sham , a TBI, and a TBI+NGF group.The rats in the TBI +NGF group were treated with NGF intranasally at 12 and 24 hours after TBI.The levels of IL-1βand TNF-αin the injured cerebral cortex were detected by ELISA , the DNA-binding activity of NF-κB evaluated by EMSA , and the expres-sion of amyloid-β( Aβ42 ) determined by Western blot . Results NGF attenuated the inflammation following TBI .Compared with the TBI group, the level of IL-1βwas obviously decreased in the TBI +NGF group at 12 hours (70.65 ±3.10 vs 37.51 ±1.92) and 24 hours (68.85 ±8.10 vs 36.23 ±2.99, P<0.05), and so was that of TNF-α(47.12 ±7.38 vs 27.63 ±5.77 and 56.15 ±11.20 vs 29.94 ±8.62, P<0.05).The DNA-binding activity of NF-κB was reduced to 111.62 ±0.49 and 131.52 ±0.88, and the expression of Aβ42 to 0.23 ±0.008 and 0.52 ±0.004 at 12 and 24 hours respectively after treatment with NGF , both with statistically significant differences from the TBI group (P<0.05). Conclusion Intranasal administration of NGF attenuates TBI-induced neuroinflamma-tion in rats, which may be associated with its regulatory effect on the Aβ42/NF-κB signaling pathway .

Objective:With proteomic approach,to find the differentially expressed proteins spots in the peripheral blood mononuclear cells(PBMCs) of healthy people and intracerebral hemorrhage patients with liver yang forming wind syndrome(LYFWS) treated by Zhenganxifeng decoction(ZGXFD) to investigate the possible proteins of LYFWS and the cure mechanism of ZGXFD.Methods: 10 intracerebral hemorrhage patients with LYFWS were treated by ZGXFD for 7-10 days.10 healthy people were selected taken as healthy controls.10ml venous blood was drawed from every subjects and PBMCs were isolated from blood by using lymphocytes separation medium.The total proteins was extracted from PBMCs.The total proteins from either intracerebral hemorrhage patients or healthy controls were prepared by means of immobilized pH gradient based on two-dimensional gel electrophoresis.After Coomassie brilliant blue G250 staining,gel-image analysis was performed by PDQuest.The differentially expressed proteins spots were identified by matrix-assisted laser desorption ionization-time-of-flight mass spectrometry(MALD I-TOF-MS).Results: Gel-image analysis revealed that there were 22 proteins spots expressed differentially.20 proteins spots were expressed differentially in the intracerebral hemorrhage patients with LYFWS as compared with healthy control,of which 8 proteins were down-regulated,7 proteins were up-regulated and 5 proteins were lost.After treated by ZGXFD,4 proteins were down-regulated,5 proteins were up-regulated and 2 proteins were lost,9 proteins weren’t expressed di erentially as compared with healthy control.8 of 20 di erentially expressed proteins spots in the intracerebral hemorrhage patients with LYFWS identi ed by MALDI-TOF-MS as compared with healthy control.These proteins are related to cell metabolism,signal transduction and so forth,of which 6 proteins were down-regulated,1 proteins were up-regulated and 1 proteins were lost.After treated by ZGXFD,2 proteins were down-regulated,1 proteins were lost,5 proteins weren’t expressed di erentially as compared with healthy control.Conclusion: 2-DE pro les of intracerebral hemorrhage patients with LYFWS in PBMCs had been established.The proteins of intracerebral hemorrhage patients with LYFWS had been identi ed,which were related to cell metabolism,signal transduction and so forth.This research presented that ZGXFD could adjust multiple di erentially expressed proteins of intracerebral hemorrhage patients with LYFWS.It was also indicated that the cure mechanism of ZGXFD was to regulate multitarget proteins.

Objective The population -based study was conducted in Hebei Renqiu 60 -70 years old resi-dents to evaluate cardiovascular disease morbidity risk in next 10 years,to analyze the related risk factors and provide an objective basis for the prevention and control of cardiovascular diseases in the local areas.Methods Cluster sam-pling method was used to select elderly residents (aged from 60 to 70 years)in Renqiu city as the research subjects. A total of 5 010 cases,2 163 males,2 847 females were included in this study.Investigation contents included health questionnaire,anthropometric and laboratory detection.According to the Framingham Risk Score (FRS),10 -year risk probability of cardiovascular disease was calculated.Research subjects were divided into low risk group (<10%),medium risk group (≥10% and <20%),high -risk group (≥20%),and logistic regression was used to analyze the risk degree of the related risk factors.Results The 10 -year cardiovascular risk median value of 60 -70 years elderly residents was 8.0 (4.0 -16.0),among them male was 16.0 (12.0 -20.0),female was 4.0 (2.0 -6.0).Low risk group had 2 784 cases,accounted for 55.7%;medium risk group had 1 452 cases,accounted for 29.0%,high-risk group had 774 cases,accounted for 15.3%.The most important risk factors were smoking and cholesterol increasing,who contributes elderly residents aged 60 to 70 years in Renqiu cardiovascular 10 -year risk.The history of stroke,excessive drinking,waist circumference,diastolic blood pressure,elevated uric acid level and HOMA -IR were the independent risk factors in moderately high risk 10 -year probability of cardiovascular.Conclusion The elderly residents aged 60 to 70 years in Renqiu has higher cardiovascular risk of 10 years.Quitting smoking,limitting alcohol,controlling of blood pressure,blood lipids,uric acid,waist circumference and HOMA -IR are important to improve cardiovascular 10 -year risk.Among them,quit smoking and alcohol limit are the most effective measures.

One influenza H3N2 virus, A/swine/Shandong/3/2005 (Sw/SD/3/2005), was isolated from pigs with respiratory disease on a farm in eastern China. Genetic analysis revealed that Sw/SD/3/2005 was a triple-reassortant virus with a PB2 gene from human-like HIN1, NS from classical swine H1NI, and the remaining genes from human-like H3N2 virus. These findings further support the concept that swine can serve as reservoir or mixing vessels of influenza virus strains and maintain genetic and antigenic stability of viruses. Furthermore, we have successfully established a reverse genetics system based on eight plasmids and rescued Sw/SD/3/2005 through cell transfection. HI tests and RT-PCR confirmed that the rescued virus maintained the biological properties of the wild type Sw/SD/3/2005. The successful establishment of the reverse genetics system of Sw/SD/3/2005 will enable us to conduct extensive studies of the molecular evolution of H3N2 influenza viruses in swine.

Objective To express recombinant chicken interleukin 2 (IL-2) in Pichia yeast eukaryotic cell expression system for a massive preparation of chicken IL-2. Methods Spleen lymphocytes taken from Xiaoshan chicken were activated by ConA in vitro. Chicken IL-2 gene was amplified and cloned into pPIC9. The pPIC9-IL-2 was transformed into Pichia yeast by electroporation. The recombinants were induced by methanol for the expression of recombinant protein. Results The protein was examined with SDS-PAGE and Western bloting. A specific molecular weight band at about 30 ku appeared. Conclusion The recombinant protein of chicken IL-2 has been expressed in Pichia yeast.

Objective To evaluate the improvement of lower urinary tract symptoms in benign prostatic hyperplasia (BPH) patients with bladder calculi by lithotripsy and adjuvant traditional Chinese decoction. Methods A total of 72 BPH patients with bladder calculi were recruited and randomly divided into the non-adjuvant treatment group (37 patients) and the adjuvant treatment group (35 patients). The adjuvant treatment group received adjuvant traditional Chinese decoction from 2 days before lithotripsy for 2 weeks. The maximum urinary flow rate (Qmax) , residual urine volume (RU), International Prostate Symptom Score (IPSS) and Quality of Life (QOL) were assessed before and after the treatment. The time of urine routine returned to normal and the indwelling time of catheter were compared between two groups. Urinary incontinence and recurrent bladder calculi were followed up for 6 months. Results After the treatment, the score of the IPSS (12.9 ± 4.5 vs. 15.7 ± 3.9;t=2.826, P=0.006) and the RU (47.3 ± 9.2 ml vs. 58.4 ± 11.3 ml;t=4.556, P<0.001) in the adjuvant treatment group were significantly lower than those in the non-adjuvant treatment group, and the Qmax (30.4 ± 4.7 ml/s vs. 21.4 ± 3.9 ml/s;t=8.862, P<0.001 ) was significantly higher. The indwelling time of catheter (5.7 ± 2.1 d vs. 8.1 ± 2.2d;t=4.730, P<0.001) and the time of urine routine returned to normal (6.9 ± 2.3 d vs. 10.2 ± 3.1 d;t=5.106, P<0.001) in the adjuvant treatment group were significantly shorter than those in the non-adjuvant treatment group. The 6-month follow-up showed that the incidence of urinary incontinence (2.9% vs. 18.9%;χ2=4.698, P=0.030) and recurrent bladder calculi (5.7% vs. 24.3%;χ2=4.813, P=0.028) in the adjuvant treatment group were significantly shorter than those in the non-adjuvant treatment group, and the total effective rate was significantly higher (62.9%vs. 29.7%; χ2=6.672, P=0.011). Conclusions Lithotripsy and adjuvant traditional Chinese decoction can reduce the IPSS score and RU, increase Qmax, decrease urinary incontinence and recurrent bladder calculi, and improve lower urinary tract symptoms in BPH patients with bladder calculi.

Objective To assess the efficacy and safety of dl-3-butylphthalide on the treatment of acute ischemic stroke. Methods A total of 197 patients who were in the period of 72 hours of first attack of ischemic stroke of internal carotid artery with NIHSS from 5 to 25 scores were enrolled in this multi-center, randomized, double-blind and aspirin-control study. Compound " Dan Shen" was used as a baseline therapy. Results Basical recovery plus significant improvement was seen in 74.7% of the patients in dl-3-butylphthalide group and 60.9% in aspirin group (CMH value 4.0,P=0.047);There was a significant improvement for dl-3-butylphthalide group regarding NIHSS total score, total score difference value and Barthel index on the day 11th and 21st after treatment compared with control group. The main adverse reaction of dl-3-butylphthalide was increased aminotransferase and mainly the slight increase of aspartate aminotransferase, by 4.34% and 0 respectively. Conclusion dl-3-butyiphthalide should be regarded as an effective and safe treatment for ischemic stroke and a treatment without severe side effects.