Objective To investigate precise and regulable animal model of benign bile duct strictures.Methods Mini pigs were divided into four groups as sham surgery group, conventional surgery group, small caliber group and large caliber group according to random number table, with 6 pigs in each group.For the small caliber group, outside diameter 3.0mm catheter were inserted into bile duct and partial sutured.For the large caliber group, outside diameter 6.0mm catheter were inserted into bile duct and partial sutured.Conventional surgery group were treated with 5-0 absorbable suture on distal bile duct and controlled stricture severity visually.For sham surgery group, bile duct were exposed without suture.Observe the general condition of four groups.Test liver function on 1st, 3rd, 7th, 14th, 21st and 28th day postoperative.Perform cholangiography on 30th day postoperative.All four groups were put to death to observe the histological changes of liver and bile duct.Results Liver function of sham surgery group showed no obvious change during observation.On 28th day postoperative, compared to conventional surgery group, the liver enzymes ALT, AST, γ-GT, ALP, bilirubin level of small caliber group elevated(P<0.05)and the liver enzymes ALT, AST, γ-GT, bilirubin level of large caliber group elevated(P<0.05),while ALP level lowered(P<0.05).Cholangiography showed bile duct stenosis of different degree on 6 pigs of small caliber group with average diameter 2.7mm (from 2mm to 4mm).As for large caliber group, 5 pigs showed stenosis with average diameter 5.5mm (from 5mm to 7mm).3 pigs of conventional surgery group showed no obvious bile duct stenosis.The 6 pigs of sham surgery group showed no bile duct stenosis.According to designed stenosis standard, the success rate of small caliber group and large caliber group are both higher than conventional surgery group.(P<0.05).30 days after surgery, all 6 pigs of small caliber group had postoperative intraperitoneal adhesion, hepatomegaly, obvious expansion of intrahepatic bile duct and gallbladder expanded by 4~5 times.5 pigs of large caliber group had hepatomegaly, slight dilation of intrahepatic bile duct, obvious expansion of gallbladder, clear choledochal edema, sutured tissue thickening and visible bile duct stenosis.HE staining with the small caliber group showed hepatocyte spotty degenerative necrosis, expanded cholestatic intrahepatic small bile duct, swelling hepatocyte and fibrous septa.HE staining with the large caliber group showed intrahepatic small bile duct slightly expanded and hepatocyte arranged basically normal.VG staining with the small caliber group showed excessive accumulaton of collagen fibers outside bile duct mucous cells, randomly and densely arranged and irregularly shaped.VG staining with the large caliber group showed increased collagen fibers in bile duct submucosa, densely and banded arranged.Conclusion This animal model of mini pig is an relatively ideal large animal model of biliary stricture with high surgical success rate and few postoperative complications.

Objective To investigate whether the signaling pathway of Akt/protein kinase B (PKB)is involved in the pathogenesis in the early stage of chronic allograft nephropathy(CAN). Methods The kidneys of Fisher(F344)rats were orthotopically transplanted into Lewis recipients. Animals were harvested respectively at 4th,8th,12th and 16th week after transplantation for renal function and histological examination.The expression of p-Akt and MMP-9 was detected by immuno- histochemical assay,and that expression of p-Akt mRNA by RT-PCR.Results At the 16th week after kidney transplantation,the levels of 24-h proteinuria and serum creatinine in transplant rats were higher significantly than those in LEW controls and F344 controls.The expression of p-Akt protein and mRNA was up-regulated,and there was a significantly negative correlation between the expression of p-Akt and the quantity of smooth muscular cells(SMCs)in arteriolar wall,the quality of tubuloin- terstitial infiltrated mononuclear cells and also MMP-9 level.Conclusion The up-regulation of p-Akt played an important role in the pathogenesis such as proliferation and migration of SMCs,infiltrated mononuclear cells in tubulointerstitial in CAN,and may be the signaling pathway leading to the up- regulation of MMP-9 expression in the early stage of CAN.

Background Age-related cataract is one of the common causes of blindness.Although the pathophysiology of age-related cataract is far from clearly understood,it is well accepted that DNA damage plays an important role in the disease pathogenesis.Objective The purpose of this study was to quantitatively evaluate the DNA damage in peripheral lymphocytes of age-related cataract.Methods A cross-sectional study was carried out.This study complied Declaration of Helsinki and approved by Ethic Committee of Affiliated Hospital of Nantong University.Written informed consent was obtained from each subject.Two hundred and eleven patients with agerelated cataract and 147 normal subjects were enrolled from a “ Jiangsu Eye Study:Funing 2011 Eye Disease Epidemic Survey”.All the subjects aged from 50 through 80 years with matched age and gender between the two groups.The percentage of tail DNA and Olive tail moment (OTM) were detected by comet assay to assess the extent of DNA damage in peripheral lymphocytes.Statistical analyses were performed with SPSS 17.0 software,and the differences of the percentage of tail DNA and OTM were compared between the age-related cataract group and normal control group by independent sample t test as well as among the 50-59 years group,60-69 years group and ≥70 years group by one-way analysis of variance.Results Comet assay showed a round lymph cell with the clear border in the normal group;while in the age-related cataract group,the cell was bigger with a comet-like tail.The percentage of tail DNA and OTM in peripheral lymphocytes were (21.75 ± 3.51) ％ and 6.54 ± 1.65 in the age-related cataract group,and those in the normal control group were (9.31 ±3.60)％ and 2.18 ± 1.10,respectively,with significant differences between them (t =32.67,P =0.00 ; t =28.02,P =O.00).In the 50-59 years subgroup of the age-related cataract group,the percentage of tail DNA and OTM in peripheral lymphocytes were (20.04±2.86) ％ and 5.92± 1.14,and in the 60-69 years subgroup of the age-related cataract group,the percentage of tail DNA and OTM in peripheral lymphocytes were (20.77 ±2.93) ％ and 6.13 ± 1.14,which were significantly reduced in comparison with (22.79 ± 3.67)％ and 6.95±1.91 of the ≥70years subgroup(TailDNA％:q=2.75,P=0.00; q=2.02,P=0.00;OTM:q=1.03,P =0.02 ; q =0.82,P =0.00).Conclusions The pathogenesis and development of age-related cataract probably is associated with DNA damage.

Objective To explore the distribution of water with high level arsenic and prevalence of arsenism along Huai'he River and the surrounding area of Hong'ze lake in Huai'an of Jiangsu. Methods Wate rsamples were collected and tested in 2008 from 18 villages of 6 towns according to history data in 3 counties like Xuyi,Jinhu and Hongze. Samples having arsenic level higher than 0.05 mg/L were investigated by epidemiological method and the patients were diagnosed by Standard of Diagnosis for Endemic Arsenism. Results All 5199 water samples were determined,and 260 water samples were exceeding the national drinking water quality level (0.05 mg/L) in 3 counties,the rates of exceeding diagnosis were 5.6%(247/4454),0.7%(4/597),6.0%(9/148) respectively. Total detected rate of endemic arsenic disease was 5.94%(128/2155). The detected rates of age group of 0 ～ ,20 ～,30 ～ ,40 ～ ,50 ～ ,60 ～ ,70 ～ ,80 ～ were 2.86%(1/35),2.11%(2/95),1.26%(3/239),3.10%(16/516),5.53% (32/579),10.07%(41/407),11.84%(27/228),10.71%(6/56) respectively. The detected rate of male (9.10%,78/857) was higher than that of female(3.85%,50/1298,χ~2 = 25.46,P < 0.01). Conclusions Huai'he River and the surrounding areas of Hong'ze lake like Xuyi,Jinhu and Hongze are identified existing endemic arsenic disease area. The prevention of arsenism should be strengthened in these areas.

This study was aimed to investigate the killing activity of cytokine-induced killer (CIK) cells after being incubated with autologous tumor cell lysate-pulsed dendritic cells (DC) and to evaluate the clinical efficacy and side effect of autologous tumor cell lysate-loaded DC in combination with CIK on relapsed or refractory non-Hodgkin's lymphoma (NHL). Peripheral blood mononuclear cells (PBMNC) were isolated from 9 patients with NHL, and cultured with granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin-4 (IL-4) to produce DC. The DC were pulsed with autologous tumor cell lysate. T lymphocytes from PBMNC were cultured with interferon-gamma (IFN-gamma), IL-2, CD3-moAb, and IL-1alpha to prepare CIK. After receiving the immunotherapy of DC and CIK, immunologic and clinical responses were evaluated. The results showed that the AgNOR, CD3(+)CD8(+) and CD3(+)CD56(+) ratio were markedly improved after the immunotherapy (p < 0.01); IFN-gamma and IL-12 levels in supernatant of DC-CIK group were higher than that in CIK group (p < 0.01); Tumor size were significantly decreased after the immunotherapy (p < 0.05). Except transient fever and chill, no remarkable adverse event happened during or after the treatment. It is concluded that the autologous tumor cell lysate-pulsed DC in combination with CIK show ability to specifically kill the lymphoma cells, obviously increases the IS value of Ag-NOR in peripheral lymphocytes, secretes cytokines higher than CIK cells alone. This combination displays the short-term satisfied efficacy on NHL through inducing specific antitumor immunity, and can be used as an effective adjuvant measure for the routine therapy of NHL.
Adult ; Aged ; Cytokine-Induced Killer Cells ; immunology ; Dendritic Cells ; immunology ; Female ; Humans ; Immunotherapy ; Immunotherapy, Adoptive ; Lymphoma, Non-Hodgkin ; immunology ; therapy ; Male ; Middle Aged ; Recurrence ; Treatment Outcome

AIM:To investigate the effects of ubiquitin-specific peptidase 9, X-linked (USP9X) down-regula-tion on apoptosis and invasion ability in gastric carcinoma cells, and to explore its possible molecular mechanisms. METH-ODS:USP9X small interfering RNA (siRNA) and control siRNA were used to be transfected into gastric carcinoma AGS cells. The cells were divided into 3 groups, including untreated AGS group, control siRNA group and USP9X siRNA group. The expression of USP9X at mRNA and protein levels in the AGS cells with different treatments was determined by real-time PCR and Western blot. The cell viability was analyzed by CCK-8 assay. Flow cytometry and Boyden chamber were employed to examine the apoptosis and invasion ability of the AGS cells. RESULTS:USP9X siRNA significantly down-regulated the expression of USP9X at mRNA and protein levels in the AGS cells. Down-regulation of USP9X markedly induced apoptosis and reduced invasion ability of the gastric carcinoma AGS cells. Notably, down-regulation of USP9X sig-nificantly reduced the protein expression of Mcl-1 and MMP-2, but markedly increased the protein level of Bax. CON-CLUSION:USP9X may be a key regulator for apoptosis and invasion in gastric carcinoma.

OBJECTIVETo observe the changes of vascular endothelial functions and general neuro-endocrine-immunity (NEI) network under the state of qi-deficiency syndrome induced by excessive idleness and to approach their internal relevance and illuminate initially the pathophysiological mechanism of vascular lesion induced by excessive idleness.

METHODSA total of 100 male Wistar rats were randomly divided into the control group and the qi-deficiency syndrome model group, 50 rats in each group. The qi-deficiency syndrome model was established by feeding the animals with hyper-alimentation diet in combination with restricting movement for 10 weeks. Changes of common chemical signal molecules related to NEI and vascular endothelial functions were measured by the end of the experiment. Furthermore, their internal relevance was analyzed by the method of canonical correlation analysis.

RESULTSThe vascular endothelial structure and function were obviously injured in the model group. Compared with the control group, the chemical signal molecules, such as 5-hydroxytryptamine (5-HT), corticosterone (CORT), triiodothyronine (T3), tetraiodothyronine (T4), angiotensin II (Ang II), interleukin-1 (IL-1), and tumor necrosis factor-alpha (TNF-alpha) in peripheral blood of the model group (n=43) were changed significantly (P<0.05 or P<0.01). Canonical correlation analysis showed that vascular endothelial dysfunction was correlated to the changes of these signal molecules in the NEI network.

CONCLUSIONSComfort-based lifestyle induced not only vascular endothelial dysfunction but also an imbalance of the NEI network. Vascular endothelial dysfunction and the imbalanced NEI network interacted with each other, and an imbalance of the NEI network may be the pathophysiologic basis for the genesis and development of vascular endothelial dysfunction, even diseases of the blood vessel.

Animals ; Aorta ; metabolism ; pathology ; physiopathology ; ultrastructure ; Biomarkers ; analysis ; metabolism ; Cardiovascular Diseases ; etiology ; metabolism ; pathology ; physiopathology ; Disease Models, Animal ; Endothelins ; metabolism ; Endothelium, Vascular ; metabolism ; pathology ; physiopathology ; Immune System ; metabolism ; pathology ; physiology ; Male ; Neuroimmunomodulation ; physiology ; Neurosecretory Systems ; metabolism ; pathology ; physiology ; Nitric Oxide ; metabolism ; Qi ; Rats ; Rats, Wistar ; Sedentary Lifestyle ; Syndrome ; Yin Deficiency ; etiology ; metabolism ; pathology ; physiopathology

Objective:To investigate the effect of lipopolysaccharide(LPS)on the crawling ability of fibroblasts derived from the scrotal skin under hypoxic conditions,its impact on vascular regeneration during wound healing,and the potential mechanism un-derlying chronic non-healing scrotal wound caused by gram-negative bacterial infection.Methods:We extracted peripheral blood buffy coats from healthy donors after venous blood centrifugation,divided them into four groups(normoxia,normoxia+LPS,hypoxia,and hypoxia+LPS)for pretreatment,and isolated the corresponding cultured supernatant.Then we measured the levels of VEGF-A,FGF and IL-8 secreted using enzyme-linked immunosorbent assay(ELISA),determined the cytotoxicity of different pretreated superna-tants by lactate dehydrogenase(LDH)release assay,and detected the cell migration ability and vascular formation in human-umbical vein endothelial cells(HUVEC).Results:Under hypoxic conditions,the expression of HIF-1 α was significantly lower in the fibro-blasts from the chronic scrotal wound than in the normal fibroblasts(0.09±0.00 vs 0.48±0.01,P＜0.05),the viability of the cells showed no statistically significant difference from that under normoxic conditions(P＞0.05),and the proliferation and migration abilities of the cells were markedly improved([83.09±2.56]％vs[17.84±1.12]％,P＜0.05).The viability of the scrotal fibroblasts was significantly lower in the normoxia+LPS than in the hypoxia+LPS group([20.15±1.19]％vs[32.54±1.57]％,P＜0.05).The cells treated with LPS,compared with those in the non-LPS groups,exhibited remarkably decreased endo-thelial cell tubes formed(5.20±0.15 va 55.30±1.15,P＜0.05),down-regulated expression of VEGF(OD value:0.59±0.07 vs 1.25±0.11,P＜0.05)and up-regulated expression of IL-8(OD value:2.42±0.07 vs 0.87±0.11,P＜0.05).Conclusion:The supernatant derived from the peripheral blood buffy coats treated with LPS has a stronger cytotoxicity and weaker an-giogenesis than that untreated under hypoxic or normoxic conditions,which may be one of the critical factors for difficult healing of scrotal wound.

The bZIP (basic leucine zipper) gene family is one of the largest transcription factor families in eukaryotes, and its members play important roles in stress response, secondary metabolism, plant growth, seed development and other aspects. To investigate the biological functions of the bZIP (CsbZIP) gene in Cannabis sativa L., we systematically investigated the CsbZIP gene family using bioinformatics methods based on the whole-genome and transcriptome data. The results showed that 55 CsbZIP gene family members (CsbZIP1-CsbZIP55) were identified and distributed on 10 chromosomes, belonging to 12 subfamilies. The gene structure and protein motif distribution of the same subfamily members were similar. Segment repeats were the main reasons for the expansion of CsbZIP gene family. Cis-elements analysis showed that the promoter regions of 73 lipid synthesis genes contained G-box or A-box element. qRT-PCR showed that the relative expression levels of 7 CsbZIP genes and 7 lipid synthesis genes were relatively high in hemp seed. 7 CsbZIP genes had a significant positive correlation with 7 lipid synthesis genes. This study revealed the structural features, evolutionary patterns and expression patterns of CsbZIP, providing important clues for further study on the regulation of CsbZIP on oil metabolism of hemp seed.

OBJECTIVE: To analyze the report status of outcomes and measurement instruments of randomized controlled trials (RCTs) of acupuncture for post-stroke dysphagia, so as to provide a basis for designing clinical trials and developing the core outcome set in acupuncture for post-stroke dysphagia. METHODS: RCTs of acupuncture for post-stroke dysphagia were searched in databases i.e. CNKI, SinoMed, Wanfang, PubMed, EMbase, Web of Science and clinical trial registries i.e. ClinicalTrials.gov and Chinese Clinical Trial Registry (ChiCTR), from January 1st, 2012 to October 30th, 2021. By literature screening and data extraction, outcomes and measurement instruments were summarized and analyzed. RESULTS: A total of 172 trials (including 165 RCTs and 7 ongoing trials registrations) were included, involving 91 outcomes. The outcomes could be classified into 7 domains according to functional attributes, namely clinical manifestation, physical and chemical examination, quality of life, TCM symptoms/syndromes, long-term prognosis, safety assessment and economic evaluation. It was found that there were various measurements instruments with large differences, inconsistent measurement time point and without discriminatively reporting primary or secondary outcomes. CONCLUSION The status quo of outcomes and measurement instruments of RCTs of acupuncture for post-stroke dysphagia is not conducive to the summary and comparison of each trial's results. Thus, it is suggested to develop a core outcome set for acupuncture for post-stroke dysphagia to improve the normative and research quality of their clinical trial design.
Humans ; Deglutition Disorders/therapy* ; Randomized Controlled Trials as Topic ; Acupuncture Therapy ; Databases, Factual ; Physical Examination ; Stroke/complications*