Objective To establish a method for qualitative detection of the presence or absence of all KIR genes by quantitative polymerase chain reaction(Q-PCR).Methods Based on the polymorphism of high-resolution level KIR alleles in Chinese population and the IPD-KIR database,KIR gene-specific primers were designed to amplify all the 16 KIR genes and 2DS4-Normal and 2DS4-Deleted subtypes by Q-PCR.Meanwhile,one negative control and one positive control specific amplifying human growth hormone(HGH)gene fragment were set to monitor the false positive and false negative results in PCR amplification,respectively.A total of 302 samples with known KIR genotype previously identified by KIR PCR-SSP commercial kit were randomly selected for blind inspection to verify the reliability of KIR Q-PCR method established by au-thors.Results The results of 300 samples detected by our KIR Q-PCR method were consistent with the known results,but two samples showed inconsistent results.One sample was negative for 2DS5 by Q-PCR but positive by PCR-SSP,another sample was positive for 2DS1 by Q-PCR but negative by PCR-SSP.The two doubtful samples were genotyped by sequencing-based typing(PCR-SBT)for 2DS5and2DS1,respectively.PCR-SBT results confirmed that the results of Q-PCR test was correct.Conclusion The KIR Q-PCR method established in this paper can provide accurate and reliable results for testing the presence or absence of KIR genes.

Objective:To design and develop a portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma,which aimed at a scene of rescue environment at frontline,and its feasibility was evaluated preliminarily through animal experiment.Methods:Based on the actual demands of the rescue environment at frontline,a set of portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma(abbreviation:minimally invasive diagnostic and therapeutic device)was researched,developed and assembled,which included portably integrated host,disposable flexible lens of endoscope,disposable apparatus of minimally invasive surgery,extendable channel device of avoiding pneumoperitoneum and so on.A male Bama miniature pig was selected,and it received two different surgeries included portably minimally invasive diagnostic and therapeutic device,and conventionally laparoscopic surgery after it received general anesthesia.The damage controls included hemostasis of intraoperative parenchyma organ,sealing and repairing of gastrointestinal perforation and drainage of indwelling catheter in abdominal and pelvic cavity between two groups were compared,and the difference of the mobility performance between them also was compared.The operational evaluation of minimally invasive surgery of damage control surgery and the potential of its clinical conversion were conducted.Results:Compared to conventional laparoscopy,this minimally invasive diagnostic and therapeutic device had better mobility,and the transfer time of this device was(3.3±1.0)min,which was significantly shorter than(14.5±3.2)min of conventional laparoscopy,and the difference of that between two device was significant(t=-5.786,P<0.05).The minimally invasive diagnostic and therapeutic device could successfully realize a series of operation of damage control surgery included exploration,flushing,suction,hemostasis,repair and drainage under the pneumoperitoneum or without pneumoperitoneum,which operation was safety and feasibility.Conclusion:The portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma can realize integration and optimization,and mobility and portability on the basis of the current laparoscopic platform,which can successfully realize the operation of damage control surgery.It has favorable application prospects and capabilities of clinical conversion.

Objective:To investigate the risk factors of pathological complete response(pCR)after neoadjuvant therapy for locally advanced gastric cancer(LAGC)and assess the value of gastric tumor markers for predicting pCR in LAGC patients.Methods:We retrospectively ana-lyzed the clinical and pathological characteristics of 213 patients who underwent radical gastrectomy and gastric tumor marker analysis after neoadjuvant therapy at The Chinse PLA General Hospital First Medical Center,between January 2020 and April 2024(20 and 193 cases in the pCR and non-pCR groups,respectively).The interrelationships among pCR,tumor markers,and clinicopathological features were compared,and independent risk factors for pCR were analyzed.A nomogram was constructed to predict the pCR.Results:Among 213 patients,20(9.4% )achieved pCR.Univariate analysis showed that age(P=0.067),tumor bed diameter(P<0.001),gastrin-17 levels(P=0.005),CA72-4 levels(P=0.073),pepsinogen ratio(P=0.024),and neoadjuvant immunotherapy(P=0.022)were strongly associated with pCR in LAGC pa-tients.Multivariate analysis showed that neoadjuvant immunotherapy,CA72-4 levels<2.5 U/mL,gastrin-17 levels<1.48 pmol/L,and tumor bed diameter<2.85 cm were independent predictive factors for pCR in LAGC patients(P<0.05).These indicators were incorporated into a nomogram prediction model;an receiver operating characteristic curve(ROC)was plotted with an AUC(95% CI)of 0.863(0.785-0.942).The calibration and decision curves suggested that the nomogram was well calibrated and had a good net benefit.Conclusions:Gastric tumor markers can effectively predict pCR after neoadjuvant therapy in LAGC patients.Our nomogram showed a good predictive ability for pCR.Thus,our findings can serve as a useful reference for clinical decision making for LAGC patients.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Pregnancy with autoimmune diseases is a challenging area that involves multidisciplinary management of immunology, rheumatology, and obstetrics. During pregnancy, the maternal immune system dynamically regulates the balance between maintaining immune tolerance to the fetus and autoimmune. However, for women with common systemic autoimmune diseases such as systemic lupus erythematosus, antiphospholipid syndrome, and rheumatoid arthritis, this balance can be upset, leading to exacerbation or recurrence of the disease and significantly increasing the risk of pregnancy complications. This paper describes the adaptive changes of the immune system during pregnancy and the effects of systemic autoimmune diseases on pregnancy outcomes, proposes multidisciplinary management strategies and challenges in perinatal pregnancy, and aims to provide references for optimizing clinical practice and improving pregnancy outcomes.

Objective:To investigate the role of complement C3a receptor in the diabetic nephropathy pathogenesis of db/db mice, and to provide a new target for prevention and treatment of diabetic nephropathy.Methods:Twelve 8-week-old male mice with type 2 diabetes mellitus (db/db mice) and 6 wild-type (db/m) mice were reared in the special pathogen free environment. The mice were grouped into db/m group, db/db group and C3a receptor antagonist group, with 6 mice in each group. db/db model mice were intraperitoneally injected with C3a receptor antagonist (SB290157, 10 mg/kg) once every two days for 8 weeks in C3a receptor antagonist group. Blood and urine samples were collected, and body weight of mice, fasting blood glucose, serum creatinine, blood urea nitrogen, urinary microalbumin/urinary creatinine and urinary N-acetyl-β- D-glucosaminidase (NAG) were detected. Renal tissues were collected, and HE, PAS and Masson stainings were used to observe the pathological changes. Immunohistochemistry, immunofluorescence and Western blotting were used to detect the protein expression levels of C3 and C3a receptor. Western blotting was used to analyze the protein expression levels of kidney injury molecule-1 (Kim-1), α-smooth muscle actin (α-SMA), zonula occluden-1 (ZO-1), vimentin and E-cadherin in renal tissues. Immunofluorescence was used to analyze the protein expression levels and distribution of α-SMA, ZO-1 and Kim-1, and immunohistochemistry was used to analyze the protein expression levels of interleukin-1 (IL-1) and tumor necrosis factor-α (TNF-α). TUNEL assay was used to detect apoptotic cells in renal tissues. Results:Compared with db/m group, body weight, fasting blood glucose, urinary microalbumin/urinary creatinine and urinary NAG in db/db group were significantly higher, while these indicators in C3a receptor antagonist group were slightly lower than those in db/db group (all P<0.01). There were no significant differences in serum creatinine and blood urea nitrogen among the three groups (all P>0.01). Compared with db/m group, db/db group had glomerular hypertrophy, necrosis and exfoliation of renal tubular epithelial cells, and dilation of renal tubules, and C3 and C3a receptor protein expression levels were higher (both P<0.01). Compared with db/db group, C3a receptor antagonist group had less glomerular lesions, mild necrosis of renal tubular epithelial cells and less tubular dilation. Compared with db/m group, the protein expression levels of Kim-1, IL-1 and TNF-α in kidney tissues of db/db group were significantly higher, while Kim-1, IL-1 and TNF-α in C3a receptor antagonist group were significantly lower than those in db/db group (all P<0.01). Compared with db/m group, the protein expression levels of α-SMA and vimentin of renal tubular epithelial cells in db/db group were significantly higher, while the protein expression levels of ZO-1 and E-cadherin were significantly lower (all P<0.01). Compared with db/db group, the protein expression levels of α-SMA and vimentin of renal tubular epithelial cells in C3a receptor antagonist group were significantly lower, and the protein expression levels of ZO-1 and E-cadherin were significantly higher (all P<0.01). Compared with db/m group, the number of apoptotic cells of kidney tissues in db/db group was increased, while the number of apoptotic cells in C3a receptor antagonist group was reduced compared with db/db group. Conclusions:The expression levels of C3 and C3a receptor of kidney tissues in db/db mice are significantly increased. Antagonistic C3a receptor can reduce the body weight, blood glucose, urinary microalbumin/urinary creatinine and urinary NAG, alleviate renal pathological injury, inhibit renal tissue inflammation, apoptosis and renal tubule epithelial-mesenchymal transition in db/db mice.

OBJECTIVE: To develop a polymerase chain reaction-sequence specific primer (PCR-SSP) method for simultaneous amplification and identification of the KIR genes among Chinese population. METHODS: Peripheral blood samples from 132 healthy donors who had given blood at Shenzhen Blood Center from January 2015 to November 2015 were selected as the study subjects. Based on the polymorphism and single nucleotide polymorphism (SNP) information of high-resolution KIR alleles in the Chinese population and the IPD-KIR database, specific primers were designed to amplify all the 16 KIR genes and the 2DS4-Normal and 2DS4-Deleted subtypes. The specificity of each pair of PCR primers was verified by using samples with known KIR genotypes. During PCR amplification of the KIR gene, co-amplification the fragment of human growth hormone (HGH) gene by multiplex PCR was used as the internal control to prevent false negative results. A total of 132 samples with known KIR genotypes were randomly selected for blind inspection to verify the reliability of the developed method. RESULTS: The designed primers can specifically amplify the corresponding KIR genes, with clear and bright bands for the internal control and KIR genes. The results of detection are fully consistent with the known results. CONCLUSION The KIR PCR-SSP method established in this study can yield accurate results for the identification of the presence of KIR genes.
Humans ; Receptors, KIR/genetics* ; Reproducibility of Results ; Polymorphism, Genetic ; Genotype ; Multiplex Polymerase Chain Reaction

OBJECTIVE: To investigate the association of molecular genetic polymorphism of KIR-HLA systems with acute lymphoblastic leukemia (ALL) and acute myelocytic leukemia (AML) in southern Chinese Han. METHODS: A total number of 323 cases of adult ALL patients, 350 adult AML, and 745 random healthy controls were tested by KIR PCR-SSP and HLA-A, -B, -C sequence-based typing (PCR-SBT) methods. The molecular genetic polymorphisms of KIR genes and KIR gene profiles, classⅠ HLA ligands, and KIR receptor +HLA ligand combinations were compared between patient and healthy control groups. RESULTS: A total number of 32 and 33 different kinds of KIR profiles were identified in the ALL and AML patient groups. Compared with the observed frequencies of KIR profiles in healthy controls, the observed frequencies of KIR profile AA1 were significantly lower in both the ALL and AML groups (ALL group: 45.79% vs. 55.30%, Pc=0.004; AML group: 48.27% vs. 55.30%, Pc=0.030). In the ALL group, the observed frequencies of 2DL2 gene and 2DL2+HLA-C1 combination, 2DS2 gene and 2DS2+HLA-C1 combination were significantly higher than those in healthy controls (P<0.05), whereas the frequencies of 2DL3 gene, HLA-A3/A11 ligand and 3DL2+HLA-A3/A11 combination were significantly lower than those in healthy controls. However, no significant differences remained after Bonferroni correction (Pc>0.05). In AML group, the observed frequencies of both 2DS1 and 2DL5 genes were significantly higher than that in healthy controls, whereas the frequencies of HLA-C2 ligand and 2DL1+HLA-C2 combination were significantly lower than that in healthy controls(P<0.05). However, no significant difference existed after Bonferroni correction (Pc>0.05). CONCLUSION This study revealed some potential susceptibility or protective factors related to acute leukemia in southern Chinese Han, especially the protective factor KIR profile AA1, which might provide new clues and theoretical basis for the pathogenesis of acute leukemia and individualized immunotherapy.
Adult ; China ; Gene Frequency ; Genotype ; HLA-A3 Antigen/genetics* ; Humans ; Leukemia, Myeloid, Acute/genetics* ; Ligands ; Polymorphism, Genetic ; Receptors, KIR/genetics*

Objective:To investigate the association of human leukocyte antigen (HLA) class Ⅰ (A, B and C), class Ⅱ (DRB1, DQB1 and DPB1) allelic and haplotypic polymorphisms with acute lymphoblastic leukemia (ALL), acute myeloid leukemia(AML) and chronic myeloid leukemia (CML) in Han nationality of southern China.Methods:The peripheral blood samples of 845 leukemia patients (323 cases of ALL, 350 cases of AML and 172 cases of CML) and 745 healthy blood donors from Han nationality of southern China in Shenzhen Blood Center were collected. The HLA-A, -B, -C, -DRB1, -DQB1 and -DPB1 genes were genotyped by using polymerase chain reaction-reverse sequence specific oligonucleotide (PCR-rSSO) and polymerase chain reaction-sequence based typing (PCR-SBT) methods to identify the first 4 digits of HLA alleles. The Arlequin 3.5 software was used to analyze HLA haplotypes. The correlations between HLA allelic and haplotypic polymorphisms and three types of leukemia were statistically analyzed at HLA low-resolution level (the first 2 digits of alleles) and high-resolution level (the first 4 digits of alleles), respectively.Results:P-values were adjusted by Bonferroni correction. In ALL group, the frequencies of A*02 (36.22% vs. 28.26%, χ 2 = 13.41, PC < 0.01) and its haplotype A*02-B*46-C*01 (15.35% vs. 10.23%, χ 2 = 10.90, PC = 0.02), DRB1*12 (15.79% vs. 11.10%, χ 2 = 9.02, PC = 0.03), A*02:03 (9.75% vs. 5.32%, χ 2 = 14.25, PC = 0.002) and its haplotype A*02:03-B*38:02-C*07:02 (3.80% vs. 1.51%, χ 2 = 10.41, PC = 0.02) were higher than those in healthy controls, which implied that these factors could confer risk effect in ALL. In AML group, the frequency of A*11-B*15-C*08-DRB1*15-DQB1*06-DPB1*02 (1.34% vs. 0.07%, χ 2 = 12.54, PC = 0.003) was significantly higher than that in healthy controls, suggesting that the risk effect might be conferred by this haplotype. In CML group, the frequencies of A*02 (36.63% vs. 28.26%, χ 2 = 9.33, PC = 0.02) and its haplotype A*02-B*15-C*04 (2.17% vs. 0.29%, χ 2 = 11.74, PC = 0.02), and DRB1*03:01-DQB1*02:01-DPB1*02:01 (1.86% vs. 0.14%, χ 2 = 13.10, PC = 0.01) were higher than those in healthy controls, which implied that these factors could confer risk effect in CML, whereas the frequency of DRB1*13 (1.45% vs. 5.25%, χ 2 = 9.29, PC = 0.03) was lower than that in healthy controls, suggesting that it was a CML antagonistic gene. Conclusion:Leukemia susceptible or antagonistic HLA alleles and haplotypes are found at low-resolution and high-resolution levels of HLA, which might provide reference for investigating the pathogenesis of leukemia and guiding formulation of effective treatment strategies in Han nationality of southern China.

【Objective】 To establish a blood transfusion outcome prediction model for comprehensivel evaluation of coagulation function of patients with upper gastrointestinal bleeding by thrombelastogram (TEG) and blood coagulation indicators. 【Methods】 The data of 101 patients with upper gastrointestinal hemorrhage, admitted to the Department of Gastroenterology of Zhejiang Provincial People′s Hospital and its Chun′an Branch from June 2018 to June 2021, were collected through Tongshuo blood transfusion management system and His system. Those patients were divided into blood transfusion group (n=56) and non-transfusion group (n=45), and into cirrhosis group (n=74) and non-cirrhosis group (n=27), and 40 patients, with non-upper gastrointestinal bleeding, were enrolled as the control. The results of TEG indicators (R, K, α, MA), coagulation function (PT, INR, APTT, TT, Fib), blood routine (Hb, Plt, WBC, NEUT%) and biochemical detection(Alb, SCr, ALT, AST, GGT) before transfusion were compared between groups and the correlation between TEG indicators and traditional coagulation parameters was analyzed. Single-factor and multi-factor analysis were used to screen blood transfusion-related factors to establish a predictive model. 【Results】 The comparisons of paremeters between transfusion and non-transfusion group were as follows, K (min), α (°), and MA (mm) was 3.86±3.12 vs 2.50±1.47, 54.00±14.08 vs 61.05±10.88, and 51.12±13.37 vs 58.26±11.08, respectively (P<0.01); PT (s) and Fib (g) was 16.36±7.45 vs 13.44±1.50 and 1.59±0.87 vs 2.35±1.09 (P<0.01); NEUT% and Hb (g/L) was 0.75 ±0.13 vs 0.66±0.15 and 68.04±14.49 vs 100.73±22.92 (P<0.01); Alb (g/L) and SCr (nmol/L) was 29.73±6.08 vs 33.73±7.19 and 99.50±53.55 vs 76.25±19.28 (P<0.01). Correlation analysis showed that APTT was positively correlated with R and K values, and negatively correlated with α and MA. Fib was negatively correlated with K values, and positively correlated with α and MA. Plt was negatively correlated with K values, and positively correlated with α and MA (P<0.01). Eight pre-transfusion indicators as K, MA, PT, Fib, NEUT%, Hb, Alb, and SCr were subjected to Logistic regression to establish a blood transfusion prediction model. The optimal ROC curve of blood transfusion threshold (blood transfusion predictive value of patients), sensitivity, specificity and AUC were 0.448, 92.9%, 88.9%, and 0.969, respectively. 【Conclusion】 The establishment of Logistic regression model by integrating detection indicators of TEG, coagulation function, blood routine and biochemistry in patients with upper gastrointestinal bleeding have showed significant correlation with blood transfusion prediction, and good clinical practicability.