BACKGROUND: Two-dimensional electrophoresis (2-DE) is precise and effective for the isolation and analysis of complex protein mixtures, has become one of valuable key techniques for proteome. Mice models were as research subjects to model human disease status, one of important methods for cardiovascular disease. The establishment and optimization of 2-DE for proteomes of mice myocardium will provide basis for heart disease.OBJECTIVE: To establish and optimize a stable technique of 2-DE for proteomic analysis of mice myocardium.DESIGN: Mice were used as research subjects, observational comparative study.SETTING: Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University.PARTICIPANTS: The experiment was performed at the Department of Pathology, Nanfang Hospital Affiliated to Southern Medical University between February and September 2004. Totally 12 male BABL/c mice of 4-5 weeks without special-pathogen free with the body mass of 12-15 g were selected, which was provided by the Experimental Animal Center of Southern Medical University.METHODS: The mice were put to death by dislocation of cervical vertebra under drugged state to obtain heart. Various protein extraction method,loading sampling buffer, strip range of IPG, sampling volume of protein,staining method and so on were compared, analyzed, scanned and digilized,and then performing image analysis. Each link in the study was established and optimized.MAIN OUTCOME MEASURES: Resolution of protein and reproducibility of the trial.RESULTS: A method that was optimal for cardiac tissue can isolate about a total of 910 protein spots on pH 4-7 gel strips. 200 μg was fit for silver stain while 1000 μg was for Coomassie stains. It was found that in the myocardium of normal mice about (920±30) protein spots were obtained in 17 em pH 3-10 L gel strip, while around (880±30) were gained in the pH 4-7 L gel strip. The mean matching rate achieves to 86. 9%.CONCLUSION: The findings of the trial showed that 2-DE technique can be effectively applied for proteomics of myocardium of mice to regulate and optimize, and relatively stable separation method of myocardial protein was established.

Objective To investigate the impact of graft position shift on anterior cruciate ligament reconstruction induced by femoral fixation of interference screw. Methods Nineteen fresh cadaveric knees were used and assigned to three groups. 1) Study of graft position shift: 5 knees were randomly selected, interference screws of 7 mm, 8 mm and 9 mm were used in autologous tendon fixation, then the graft position shift were measured. 2) Study of isometry: 7 knees were randomly divided into the isometric reconstruction group (D group). In the D group, Retrobutton, interference screw and interference screw in location-corrected bone tunnel were used respectively as fixation. The isometry of grafts was evaluated. 3) Study of tibia anterior translation: 7 knees were randomly divided into the anatomic reconstruction group (J group). In the J group,the tibia anterior translation was measured in four different conditions in the same joint: intact knee joint,knee joint without ACL, ACL anatomic reconstruction by interference screw fixation, and ACL anatomic reconstruction by interference screw fixation with corrected bone tunnel location. Results 1) With 7 mm, 8mm and 9 mm interference screw fixation, graft position shift were (2.36±0.11) mm, (2.72±0.06) mm and (3.00±0.06) mm respectively. 2) Graft length change: graft length change in Retrobutton group and corrected bone tunnel group were less than 3 mm, while graft length change in those fixed with interference screw were stretched in more than 3 mm. 3) Study of tibia anterior translation: there was no difference among the intact group, the anatomic group and the corrected group at 0° and 15°. However, the difference was found between the intact group and other groups at 30°、60° and 90° of flexion, as well as between these two reconstructed methods at 20° joint flexion (P＜0.05). Conclusion In both isometric and anatomic ACL reconstruction with interference screw, the graft is pushed tightly toward the femoral tunnel wall, which shifts the graft away from the desired position. In our study we find out that the corrected location of the femoral bone tunnel significantly improves the isometry of ACL reconstruction and anatomic reconstruction.

Objective To assess the value of fuorine-18-fluom deoxy glucose positron emission tomography /computer tomography (18F-FDG PET/CT) in diagnosis of lymph node metastasis in the cases with non-small cell lung cancer ( NSCLC) .Methods From March 2012 to March 2015,167 patients underwent 18F-FDG PET/CT and contrast enhanced CT inspection within 10 days prior to the surgery and were pathologically diagnosed as non-small cell lung cancer(NSCLC) after surgery.With regard to estimation of the lymphatic metastasis,we com-pared the sensitivity,specificity,accuracy,positive and negative predictive value ,and Youden index between the PET/CT and contrast en-hanced CT.Results There were 731 lymph node stations from the 167 patients.Referred to the final pathological results ,PET/CT has screened 143 lymph node stations in true positive set ,26 lymph node stations in false positive set ,61 lymph node stations in false negative set , and 501 lymph node stations in true negative set .Furthermore,the sensitivity,specificity,the positive and negative forecast values ,and Youden index of PET/CT and CT was 70.10%vs.54.19%(P<0.05),95.07%vs.92.23%(P<0.05),88.10%vs.81.67%(P<0.05),84.62%vs.72.85%(P<0.05),89.15%vs.81.67%(P<0.05) and 0.65 vs.0.46 (P<0.05),respectively.Conclusion It is more effective to to precisely validate lymphatic metastasis of NSCLC by using PET /CT than using contrast enhanced CT inspection .Additionally,PET/CT can provide more information for the preoperative diagnosis , staging and the follow-up treatment of lung cancer .

T,and 916-917 ins G were SLC26A4 mutations unreported hitherto, which may be specific to the Chinese population. CONCLUSION The EVA syndrome is a typical autosomal recessivehereditary disease caused by mutations in SLC26A4 gene. Genetic testing of SLC26A4 is the one of the important diagnostic methods for EVA syndrome.

The secondary damage after traumatic brain injury (TBI) involves a variety of pathological processes, and the inflammatory response in the nervous system is an important factor which affects nerve repair and regeneration. As the innate immune cell in the nervous system, microglia (MG) plays an important role in the entire neuroinflammatory environment by regulating the activation state of MG and changing the inflammatory response in the direction of promoting nerve repair and regeneration, which has great potential in treatment of TBI. This article reviews the inflammatory response of the nervous system after TBI and the reactivity of MG, as well as their significance in nerve repair and regeneration.

Objective:To establish a model of patellar dislocation by femoral osteotomy or surgical release of medial retinaculum in immature rabbits, and observe morphological and trabecular microarchitectural changes in the trochlea.Methods:Forty rabbits at 3 months of age were included. All right knees underwent surgery, 20 knees were treated with femoral osteotomy and internal rotation of distal femur to increase femoral anteversion angle (Osteotomy group, OS group), and another 20 knees were treated with surgical release of medial retinaculum and overlap suture of lateral retinaculum (Soft tissue group, ST group). All left knees were acting as internal controls. Micro-CT scans for distal femur were acquired after 4 months post surgery. the height of Medial, central, and lateral trochlear, sulcus angle, and lateral and medial trochlear slope were measured to describe the trochlear morphology, and bone volume fraction (BV/TV), trabecular thickness (Tb.Th), trabecular number (Tb.N), trabecular spacing (Tb.Sp), and bone mineral density (BMD) were calculated to evaluate the microarchitectural structure. All parameters were compared between groups.Results:In OS group, one rabbit sustained a hip dislocation without patellar dislocation. Three knees developed complete patellar dislocation in daily flexion position, and the remaining 16 patellae were dislocated when the knee was placed in the maximal extension position. In ST group, 15 knees were complete patellar dislocation in daily flexion position, and 5 knees were without dislocation. A local boss was formed proximal to the entrance of the groove and the articular cartilage was smooth, and no obvious osteoarthritis was observed in OS group. In ST group no boss was formed, while obvious cartilage degeneration and defect was seen. Compared to the control group, the central trochlear height and sulcus angle were greater in both groups, but without significant difference between the two groups. The Tb.Th was increased in both medial and lateral condyle, and Tb.N was decreased in medial condyle compared with its control knees in OS group. The BV/TV, Tb.Th, Tb.N and BMD were decreased and Tb.Sp was increased in both medial and lateral condyle compared with its control knees in ST group. Compared to the OS group, the BV/TV, Tb.Th, Tb.N and BMD were smaller and Tb.Sp was greater in both medial and lateral condyle in ST group, with significant differences.Conclusion:The model of patellar dislocation could be successfully achieved by femoral rotational osteotomy to increase femoral anteversion or surgical release of medial retinaculum and overlap suture of lateral retinaculum, and subsequent morphological and trabecular microarchitectural changes in the trochlea are different. Different bony and soft tissue factors should be addressed for different patients with patellar dislocation in clinical practice.

Objective: To investigate the relationship between the eosin-5′-maleimide (EMA) binding test and the clinical severity of hereditary spherocytosis (HS). Methods: A total of 258 un-splenectomize HS patients were consecutively enrolled. Correlation of hemoglobin concentration, hemolytic parameters, compensating erythropoiesis and the EMA binding test were evaluated. Results: 258 (128 male and 130 female) patients were included in this study, including 91 compensatory hemolysis patients, 53 patients with mild anemia, 78 patients with moderate anemia and 36 patients with severe anemia. The median age at diagnosis was 23 (2-70) years. The median decreased fluorescence intensity of EMA binding test was 29.97% (16.09%-47.34%) and the average intensity was (29.70±6.28) % of 258 HS patients. The decreased EMA binding fluorescence intensity correlated with MCV (r=-0.343, P<0.001) and MCHC (r=0.223, P<0.001). There was no relationship between EMA fluorescence intensity and absolute reticulocyte count (r=0.080, P=0.198) , reticulocyte percentile (r=-0.015, P=0.813) , IBIL levels (r=-0.009, P=0.902) , HGB levels (r=-0.067, P=0.280). Evaluated as a quartile variable, EMA fluorescence intensity was not correlated with anemia severity (C=0.150, P=0.746). Conclusion EMA binding test does not related to anemia levels and has no major clinical implications for disease severity in HS.

Objective: To reveal the genetic characteristics of erythrocyte membrane protein in hereditary spherocytosis (HS) in China. Methods: Next-generation sequencing technology was used to detect mutations in genes of erythrocyte membrane proteins in 51 clinically diagnosed HS patients. The relationship between gene mutations and clinical phenotypes was analyzed. Results: Mutations in erythrocyte membrane protein genes were detected in 37 patients, including 17 with ANK1 mutations (17/37, 45.9%), 14 with SPTB mutations (14/37, 37.8%), and 5 with SLC4A1 mutations (5/37, 13.5%). One patient carried both heterozygous ANK1 mutation and SPTB mutation (1/37, 2.7%). SPTA1 and EPB42 mutation was not fou nd in any patient. Nonsense mutations (36.8%) and missense mutations (31.6%) were most common. Of the 38 mutations detected, 34 were novel mutations and have not been reported elsewhere (89.5%). Sixteen HS patients underwent parental genetic validation, 6 patients (37.5%) inherited gene mutation from parents and 10 (62.5%) were de novo. The peripheral blood cell parameters of HS patients were not related to the mutant genes and gene mutation types. However, it seems that HS patients with mild clinical status are prone to carry SPTB mutations while more patients with severe clinical status have ANK1 mutations. Conclusions ANK1 and SPTB are the most common mutant genes in Chinese HS patients, mainly with missense mutations and nonsense mutations. There was no significant correlation between the mutation of HS related genes and the severity of HS.

ObjectiveTo systematically evaluate the incidence rate of low-level viremia （LLV） in chronic hepatitis B （CHB） patients and related influencing factors， and to provide evidence-based medicine evidence for effective intervention and prevention of LLV in clinical practice. MethodsThis study was conducted according to the PRISMA guideline， with a PROSPERO registration number of CRD42023455304. CNKI， Wanfang Data， VIP， SinoMed， PubMed， Embase， Web of Science， and the Cochrane library were searched for observational studies on LLV and related influencing factors in CHB patients published up to July 21， 2023. Stata 16.0 software was used to perform the meta-analysis. ResultsA total of 12 articles were included， with a total sample size of 3408 cases， among whom there were 1181 patients with LLV. The meta-analysis showed that the incidence rate of LLV was 32.8% （95% confidence interval ［CI］： 27.6%‍ ‍—‍ ‍38.3%） in treatment-experienced CHB patients. High HBsAg quantification （odds ratio ［OR］=2.107， 95%CI： 1.782‍ ‍—‍ ‍2.491， P<0.001）， positive HBeAg （OR=3.258， 95%CI： 2.629‍ ‍—‍ ‍4.038， P<0.001）， high HBV DNA level at baseline （OR=1.286， 95%CI： 1.157‍ ‍—‍ ‍1.430， P<0.001）， and history of entecavir treatment （OR=3.089， 95%CI： 1.880‍ ‍—‍ ‍5.074， P<0.001） were risk factors for LLV； duration of antiviral therapy ≥3 years （OR=0.175， 95%CI： 0.093‍ ‍—‍ ‍0.331， P<0.001） and high alanine aminotransferase level at baseline （OR=0.985， 95%CI： 0.978‍ ‍—‍ ‍0.992， P<0.001） were protective factors against LLV. The sensitivity analysis showed no significant change in effective value， suggesting that the results of the meta-analysis were relatively stable. The funnel plot of the studies included was basically symmetrical， and the results of the Egger’s test and the Begg’s test suggested that there was no obvious publication bias in the articles included. ConclusionClinicians should guide decision making based on the influencing factors for LLV and related clinical evidence， so as to reduce long-term clinical risks and avoid adverse outcomes.

OBJECTIVETo identify the characteristics of chronic natural killer cell lymphocytosis (CNKL).

METHODSThe clinical data of eight cases defined by the World Health Organization classification was retrospectively analyzed and related literatures were reviewed.

RESULTSHalf of the 8 patients were asymptomatic. Among them, the most common abnormalities were leukocytosis with the median of 11.8(4.5-20.0)×10⁹/L and high proportion of lymphocytes [0.78(0.51-0.89)], while anemia and thrombocytopenia only in 1 patient respectively. The absolute CD3⁻CD16⁺ NK cell count with the median of 5.7(2.4-9.6)×10⁹/L increased in peripheral blood. By the end of follow-up, except one case was lost, the other seven patients were in stable condition,including four cases without any medications and three patients receiving chlorambucil or glucocorticoid.

CONCLUSIONAs an indolent chronic lymphoproliferative disease, CNKL was presented with mild clinical symptoms and increased number of CD3⁻CD16⁺ NK cells in peripheral blood. Patients with symptoms could be treated with immunosuppressive therapy while those asymptomatic could be followed up without intervention.

Adult ; Aged ; Chronic Disease ; Female ; Follow-Up Studies ; Humans ; Killer Cells, Natural ; Lymphocytosis ; blood ; Male ; Middle Aged ; Prognosis