Objective:To analyze the diversity of Duffy blood group gene among ethnic Hui population from Henan Province using PacBio long-read sequencing technique.Methods:Randomly select 30 individuals with three generations of Hui ancestry from Henan as the study subjects. Full-length sequences of the Duffy blood group gene were obtained through PacBio long-read sequencing. Distribution of the predicted phenotype and genotype frequency were determined, and the linkage between Duffy haplotypes and variation sites was analyzed. Genetic diversity, natural selection pressure, and population genetic characteristics were evaluated. This study was approved by the Medical Ethics Committee of the Second Affiliated Hospital of Zhengzhou University (Ethics No. 202223).Results:The predicted Duffy blood group phenotype in the Henan Hui population was predominantly Fy(a+ b-). Three novel SNPs in the FY*01 allele were identified, with a total frequency of 13.33%, among which FY*01.NEW1 (c.199C>T) was the most common. A total of 32 variant sites were identified, with 28 located in intronic regions, indicating that genetic diversity was primarily concentrated in introns. The Duffy blood group gene was under negative selection pressure ( dN/ dS < 1, Tajima′s D, Fu and Li′s D*& F* significantly deviated from 0), suggesting overall conservation. The allele frequencies of Duffy blood group in the Henan Hui population was similar to that of the Xinjiang Hui, Xinjiang Kazakh, Inner Mongolia Mongolian, and Yuncheng Han populations, but significantly different from those of most Han and other ethnic groups ( P<0.05). Conclusion:This study revealed the characteristics of the Duffy blood group gene among the Henan Hui population and demonstrated the significant advantages of PacBio long-read sequencing technique in haplotype analysis, genetic diversity study, and novel mutation identification.

OBJECTIVE: To analyze the diversity of Duffy blood group gene among ethnic Hui population from Henan Province using PacBio long-read sequencing technique. METHODS: Randomly select 30 individuals with three generations of Hui ancestry from Henan as the study subjects. Full-length sequences of the Duffy blood group gene were obtained through PacBio long-read sequencing. Distribution of the predicted phenotype and genotype frequency were determined, and the linkage between Duffy haplotypes and variation sites was analyzed. Genetic diversity, natural selection pressure, and population genetic characteristics were evaluated. This study was approved by the Second Affiliated Hospital of Zhengzhou University (Ethics No. 2022223). RESULTS: The predicted Duffy blood group phenotype in the Henan Hui population was predominantly Fy(a+b-). Three novel SNPs in the FY*01 allele were identified, with a total frequency of 13.33%, among which FY*01.NEW1 (c.199C>T) was the most common. A total of 32 variant sites were identified, with 28 located in intronic regions, indicating that genetic diversity was primarily concentrated in introns. The Duffy blood group gene was under negative selection pressure (dN/dS < 1, Tajima's D, Fu and Li's D* and F* significantly deviated from 0), suggesting overall conservation. The allele frequencies of Duffy blood group in the Henan Hui population was similar to that of the Xinjiang Hui, Xinjiang Kazakh, Inner Mongolia Mongolian, and Yuncheng Han populations, but significantly different from those of most Han and other ethnic groups (P < 0.05). CONCLUSION This study revealed the characteristics of the Duffy blood group gene among the Henan Hui population and demonstrated the significant advantages of PacBio long-read sequencing technique in haplotype analysis, genetic diversity study, and novel mutation identification.
Female ; Humans ; Male ; Asian People/ethnology* ; China/ethnology* ; Duffy Blood-Group System/genetics* ; Ethnicity/genetics* ; Gene Frequency ; Genetic Variation ; Haplotypes ; Polymorphism, Single Nucleotide

Objective:To analyze the diversity of Duffy blood group gene among ethnic Hui population from Henan Province using PacBio long-read sequencing technique.Methods:Randomly select 30 individuals with three generations of Hui ancestry from Henan as the study subjects. Full-length sequences of the Duffy blood group gene were obtained through PacBio long-read sequencing. Distribution of the predicted phenotype and genotype frequency were determined, and the linkage between Duffy haplotypes and variation sites was analyzed. Genetic diversity, natural selection pressure, and population genetic characteristics were evaluated. This study was approved by the Medical Ethics Committee of the Second Affiliated Hospital of Zhengzhou University (Ethics No. 202223).Results:The predicted Duffy blood group phenotype in the Henan Hui population was predominantly Fy(a+ b-). Three novel SNPs in the FY*01 allele were identified, with a total frequency of 13.33%, among which FY*01.NEW1 (c.199C>T) was the most common. A total of 32 variant sites were identified, with 28 located in intronic regions, indicating that genetic diversity was primarily concentrated in introns. The Duffy blood group gene was under negative selection pressure ( dN/ dS < 1, Tajima′s D, Fu and Li′s D*& F* significantly deviated from 0), suggesting overall conservation. The allele frequencies of Duffy blood group in the Henan Hui population was similar to that of the Xinjiang Hui, Xinjiang Kazakh, Inner Mongolia Mongolian, and Yuncheng Han populations, but significantly different from those of most Han and other ethnic groups ( P<0.05). Conclusion:This study revealed the characteristics of the Duffy blood group gene among the Henan Hui population and demonstrated the significant advantages of PacBio long-read sequencing technique in haplotype analysis, genetic diversity study, and novel mutation identification.

Objective To investigate the effect and mechanism of emodin on focal cerebral ischemia in rats based on myeloid differentiation factor 88(MyD88)/extracellular signal-regulated kinase(ERK)pathway and nuclear factor-κB(NF-κB)nuclear translocation.Methods SD rats were randomly divided into sham operation group,model group and emodin group,with six rats in each group.The rat model of transient middle cerebral artery occlusion(tMCAO)was established by middle cerebral artery embolization.Rats in the emodin group were given 40 mg·kg-1 emodin by gavage for three times at 72,48 and 24 hours before modeling.At 24 hours after modeling,the neurological function of rats was scored.TTC staining was used to detect the area of cerebral infarction.HE staining was used to observe the morphological changes of brain tissue.The mRNA expression levels of MyD88 and tumor necrosis factor-α(TNF-α)in brain tissue were detected by RT-qPCR.The expression levels of MyD88,ERK,p-ERK and TNF-α in brain tissue were detected by Western Blot.The protein expression of NF-κB in brain tissue was detected by immunofluorescence.Results Compared with the sham operation group,the neurological function score of the model group was significantly increased(P＜0.01),and the cerebral infarction area was significantly increased(P＜0.01).In the cortical area of the ischemic penumbra,cell necrosis,abnormal cell morphology,nuclear fragmentation and atrophy,and the number of cells decreased significantly;the mRNA expression levels of MyD88 and TNF-α in brain tissue were significantly increased(P＜0.01,P＜0.001),the protein levels of MyD88,p-ERK/ERK and TNF-α were significantly increased(P＜0.05,P＜0.01,P＜0.001),and the proportion of NF-κB into nuclear cells was significantly increased(P＜0.001).Compared with the model group,the neurological function score of rats in the emodin group was significantly decreased(P＜0.05),and the area of cerebral infarction was significantly reduced(P＜0.05).The number and morphology of neurons in the ischemic penumbra cortex were restored to a certain extent.The mRNA expression levels of MyD88 and TNF-α in brain tissue were significantly decreased(P＜0.05,P＜0.01),the protein levels of MyD88,p-ERK/ERK and TNF-α were significantly decreased(P＜0.05),and the proportion of NF-κB into nuclear cells was significantly decreased(P＜0.001).Conclusion Emodin has a preventive and protective effect on rats with focal cerebral ischemia,which may be related to its inhibition of MyD88 activation,ERK phosphorylation and NF-κB nuclear translocation,and then down-regulation of inflammatory cascades and secretion of pro-inflammatory factors such as TNF-α,thereby exerting anti-inflammatory effects.

Objective To retrieve,evaluate and summarize the relevant evidence on malnutrition management among maintenance hemodialysis(MHD)patients to provide clinical medical staff with evidence-based guidelines for managing malnutrition in MHD patients.Methods Using the"6S"pyramid model of evidence,We searched UpToDate,BMJ best clinical practice,the Australian JBI evidence-based health care centre database(JBI),Medive,the International Guidelines Collaborative Network(GIN),Agency for Healthcare Research and Quality Network,the UK National Institute of Clinical Medicine Guidance Library(NICE),Scottish Inter-College Guide Collaboration network,the Ontario Registered Nurses Association(RNAO),Guidelines Network of International Society of Nephrology,American Nephrology Foundation Guidenet,British Nephrology Society,Society of Nephrology,Chinese Medical Association,Improving Kidney Disease Outcomes Worldwide,International Society of Renal Nutrition and Metabolism,American Society for Parenteral Enteral Nutrition,European Society for Clinical Nutrition and Metabolism,Chinese Nutrition Society,PubMed,Web of Science,Embase,Cochrane Library,CNKI,Wanfang,VIP and CBM,to collect relevant guidelines,expert consensus,clinical decision,evidence summary,recommended practice,best practice,systematic evaluation and clinical practice,from the establishment to the date of April 10th,2023,for literature on malnutrition management of MHD patients.The literature included guidelines,expert consensuses,clinical decision-making studies,evidence summaries,recommended practices,best practices,and systematic reviews.Two researchers independently evaluated,extracted,and integrated the evidence.The evidence was graded by the evidence pre-grading system of the Evidence-based Healthcare Center of Joanna Briggs Institute(JBI).Results A total of 16 articles were included and summarized,comprising 5 guidelines,3 expert consensuses,1 clinical decision-making study,1 evidence summary,and 6 systematic reviews.Thirty-nine pieces of the best evidence were integrated,covering 5 key aspects(39 evidences):screening and assessment,nutritional requirements,nutritional supplement,other interventions,and monitoring and education.Conclusions The best evidence summary of malnutrition management among MHD patients in this study is scientifically rigorous and systematic.Clinical medical staff can use the best evidence to develop personalized malnutrition management programs for MHD patients.

Objective To explore the risk factors for renal anemia in hemodialysis patients with end-stage kidney disease (ESKD). Methods A total of 48 ESKD patients undergoing hemodialysis in our hospital from December 2021 to December 2022 were selected as study objects. They were divided into renal anemia group (86 cases) and non-anemia group (62 cases) based on hemoglobin (Hb) levels and diagnostic criteria for renal anemia. Clinical data between the two groups were compared. Random forest algorithm and multivariate Logistic regression analysis were used to screen for factors influencing renal anemia, and a multivariate Logistic regression model was established. Cross-validation was also employed to verify the stability of the model. A risk stratification system was developed, and patients were stratified based on cut-off values obtained from X-Tile software. The areas under the receiver operating characteristic (ROC) curves were used to evaluate the discrimination ability of the multivariate Logistic regression model and the risk stratification system. Results Among 148 ESKD patients, the incidence of renal anemia was 58.11% (86/148). Hypertension, increased neutrophil-to-lymphocyte ratio (NLR), elevated C-reactive protein (CRP), increased serum ferritin (SF), and elevated intact parathyroid hormone (iPTH) were identified as risk factors for renal anemia (P < 0.05). Administration of α-calcidol, increased erythropoietin (EPO), elevated triglyceride (TG), and increased serum albumin (ALB) were protective factors (P < 0.05). The goodness-of-fit test for the multivariate Logistic regression model showed a Nagelkerke R2 of 0.593. Based on the cut-off values for risk scores obtained from X-Tile software, patients were stratified into low-risk group (< 4 points, 29 cases), medium-risk group (4 to < 7 points, 64 cases), and high-risk group (≥7 points, 55 cases), with renal anemia incidence rates of 24.14%, 57.81%, and 76.36%, respectively. The difference in renal anemia incidence rates among the three groups was statistically significant (P < 0.001). In the training set, the areas under the ROC curves for the multivariate Logistic regression model and the risk stratification system were 0.826 and 0.811, respectively, and were 0.804 and 0.789, respectively in the validation set. Conclusion The occurrence of renal anemia in hemodialysis patients with ESKD is influenced by various factors, including hypertension, NLR, CRP, etc.

Objective:To construct a simple model of arteriovenous fistula classification,and to achieve the classification of arteriovenous fistula in hemodialysis patients.Methods:The study was a retrospective analysis, a total of 304 hemodialysis patients with internal fistula in People′s Hospital of Deyang City from January 2016 to January 2021 were selected by convenience sampling method,depending on whether the internal fistula was dysfunctional, patients were divided into 64 in the internal fistula failure group and 240 in the internal fistula patency group. Independent influence factors and their regression coefficient were obtained by single-factor analysis and logistic regression analysis, The risk score formula was established based on the regression coefficient to form a simple model of internal fistula classification.The model was evaluated by receiver operating characteristic curve and the scoring criteria for internal fistula classification was determined.Results:Logistic regression analysis showed that diabetes mellitus, hypotension, age≥60 years old, compression time≥30 min, blood phosphorus>1.78 mmol/L, triglyceride>1.71 mmol/L and fibrinogen>4 g/L were independent influencing factors of internal fistula failure (all P<0.05).The area under the receiver operating characteristic curve was 0.858(95% CI 0.789-0.928, P<0.01), and the best critical value of the internal fistula classification was 7.5, the sensitivity was 80.4% and the specificity was 84.8%. Conclusions:By obtaining the predictors of internal fistula failure, conducted the risk score, and constructed a simple model of internal fistula classification, which can effectively predicted the risk of internal fistula failure. It is conducive to the implementation of internal fistula classification management and the puncture of corresponding grade, to ensure the pathway safety of patients.

Objective To investigate the effects and mechanism of advanced glycosylation end products (AGEs)on endothelial cell senescence and endothelial barrier dysfunction.Methods Human umbilical vein endothelial cells (HUVECs)were isolated and cultured.The cells were randomized into three groups:the control group(normal medium),the bovine serum albumin-treated group(BSA control group)and AEGs group(treated with AEGs-BSA).Senescence of HUVECs were detected by senescence-associated β-galactosidase (SA-beta-Gal)staining.The mRNA and protein expressions of senescence-related genes of p53,p21 and p16 in each group were determined by reverse transcription and real-time PCR(RT-qPCR)and Western blot.Reactive oxygen species(ROS)level was determined by dichlorodihdrofluorescence diacetate (DCFH-DA).The transendothelial electric resistance(TER)were measured by endothelial electric resistance meter.The protein levels of myosin light chain kinase (MLCK),phosphorylated myosin light chain (p-MLC),myosin light chain (MLC)were detected by Western blot.Results Compared with the control group and the BSA control group,the AGEs group showed the significantly increased positive rate of senescence-associated SA-beta-Gal staining (67.30 ± 0.75 ％ vs.7.81 ±0.35 ％ and 7.64 ± 0.91％,respectively,P ＜ 0.01)and the expressions of aging-related genes of p53,p21 and p16 were significantly increased (P ＜ 0.05)There was no significant difference in transendothelial electric resistance(TER)between the control group and theBSAgroup(48.0±6.3 Ω· cm2 vs.42.0±7.8 Ω· cm2,P＞0.05),while TER was lower in the AEGs group than in control group and the BSA group[(27.0±4.2)Ω · cm2 vs.(48.0±6.3)Ω · cm2 and (42.0 ± 7.8) Ω · cm2,P ＜0.01].ROS production had no significant difference between the control group and the BSA group[(38.36 ± 8.55) ％ vs.(41.67 ± 6.93) ％,p ＞ 0.05],while was increased in the AEGs group versus control group and the BSA group[(69.31±8.47)％ vs.(38.36±8.55) ％ and (41.67 ± 6.93) ％,P ＜0.05).The protein expression levels of MLK and p-MLC/MLC were higher in the AGEs group than in the control group and the BSA group(P＜0.05).Conclusions AGEs may lead to endothelial cell senescence and endothelial barrier dysfunction by promoting ROS production and oxidative stress,and by regulating MLCK signaling pathway.

ObjectiveTo investigate the clinical effect of the double stapling technique for anus-preserved (Dixon) and Miles procedure of rectal carcinoma. MethodsFifty-eight cases were divided into 29 cases as control group and 29 cases as observation group by digital table method. The distance of their carcinoma was 4-7 cm to anus. The control group was treated with Miles procedure of rectal carcinoma, the observation group was treated with double stapling technique for Dixon. ResultsThe operation time of observation group was ( 180 ± 56) min, the control group was (240 ± 73)min(P ＜ 0.05). Five-year survival rate of observation group was 77.9％, the control group was 79.3％ (P ＞0.05);the recurrence rate of observation group was 6.9％(2/29),the control group was 6.9％(2/29) (P ＞ 0.05). In the observation group,nobody suffered encopresis after 1 year,postoperative stool frequency was ≤5 times/d,while the control group occurred encopresis gradually. ConclusionIf the distance of carcinoma is 4-7 cm to anus, the way of the double stapling technique for Dixon can get the similar short term effect with the way of Miles, and can get more ability to control it.

To obtain a recombinant human plasminogen (hPLG) with potential anti-platelet aggregation activity, we cloned the cDNA coding Pro544 to Asn791 of hPLG, a kringle-deficit derivative (hPLG-deltaK). The Pro559 in activation loop was then mutated into Asp559 to provide Arg-Gly-Asp (RGD) motif. The constructed pPICZalphaA-RGD-HPLG-deltaK plasmid was expressed in yeast Pichia pastoris GS115, which produced RGD-hPLG-deltaK about 0.160 g/L broth. After affinity chromatography, the purity of the recombinant protein reached above 90%. Western blotting test confirmed that it retained the immunological reaction capability as human PLG. Its urokinase activation rate in 24 hours and its fibrinolytic activity made no deference against native hPLG-deltaK (P=0.630, n=5). Importantly, after activation by urokinase, RGD-hPLG-deltaK showed a significantly higher platelet aggregation inhibition rate (Ri) (21.8% +/- 1.57%) than hPLG-deltaK (3.8% +/- 0.33%) (P=0.000, n=5). These results proved that we constructed an hPLG mutant with anti-platelet aggregation activity, which made a foundation for developing innovative thrombolytic drugs with multifunction.
Humans ; Kringles ; genetics ; Oligopeptides ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Plasminogen ; biosynthesis ; genetics ; Platelet Aggregation Inhibitors ; isolation & purification ; pharmacology ; Point Mutation ; Recombinant Proteins ; biosynthesis ; genetics ; isolation & purification ; pharmacology