BACKGROUND: Cryopreservation can decrease tissue and organ immunogenicity. The effects of cryopreservation on cell immunogenicity are disputed.OBJECTIVE: To investigate the effects of liquid nitrogen cryopreservation on osteoblast immunogenicity. DESIGN: Randomized,controlled ,paired-sample experiment. SETTING: This study was performed in the Laboratory Center, Qilu Hospital Affiliated to Shandong University between July 2003 and March 2004. MATERIALS: Four New Zealand rabbits of either gender were included for this study. 3H-TdR was provided by Nuclear Medicine Institute of Shandong University. METHODS: Osteoblasts were cultured from the tibial periosteum of New Zealand rabbits and then cryopreservated in the liquid nitrogen for 3 months and defrosted. Cryopreservated and thawn osteoblasts were set as cryopreserved group and freshly cultured osteoblasts were set as non-cyropreserved group. Major histocompatibility complex (MHC)-I positive rate was examined by flow cytometry assay prior to and after cryopreservation. Simultaneously, mixed lymphocyte-osteoblast cultures were established. Lymphocyte stimulation index was calculated after counting the flares using β liquid scintilloscope. MAIN OUTCOME MEASURES: MHI-I antigen positive rate and lymphocyte stimulation index prior to and after cryopreservation of osteoblasts. RESULTS: MHI-I antigen positive rate and lymphocyte stimulation index of osteoblasts was significantly higher in the non-cryopreserved group than in the cryopreserved group (P＜0.01). CONCLUSION: The immunogenicity of cryopreserved osteoblasts was significantly decreased. Liquid nitrogen cryopreservation is an ideal method to decrease the immunogenicity of osteoblasts.

ObjectiveTo investigate the clinical characteristics and pathogenic microorganisms in culture-positive sepsis, to identify its risk factors, and evaluate the prognosis on polymicrobial infection in intensive care unit (ICU).Methods A descriptive retrospective study was conducted. Clinical data of patients aged≥ 18 years, diagnosed as culture-positive sepsis, and admitted to six ICUs of Guangdong General Hospital from October 12th, 2012 to December 1st, 2014 were enrolled. Based on the number of isolated pathogens, patients were divided into polymicrobial infection group (≥two pathogens) and monomicrobial infection group (one pathogen) to investigate the clinical characteristics of patients with culture-positive sepsis and the causative pathogens. Multiple logistic regression was conducted to identify the risk factors for polymicrobial infection. Kaplan-Meier curve was plotted to analyze a 90-day survival rate from the onset of positive blood culture.Results 299 patients with positive blood culture were enrolled. A total of 450 strains of pathogens were isolated including 246 gram-positive cocci (54.67%), 167 gram-negative bacilli (37.11%) and 37 fungi (8.22%). Ninety-one patients had polymicrobial infection, and 208 with monomicrobial infection. Compared with monomicrobial infection group, patients suffering from polymicrobial infection had more advanced age (years: 73.19±18.02 vs. 60.83±18.06,t = -5.447,P = 0.000), also with higher incidence of cerebrovascular diseases [39.56% (36/91) vs. 17.79% (37/208),χ2 = 16.261,P = 0.000] or chronic renal insufficiency [15.38% (14/91) vs. 7.21% (15/208),χ2 = 4.828,P = 0.028], higher incidence of recent hospital stay (≥2 days) within 90 days [73.63% (67/91) vs. 61.54% (128/208),χ2 = 4.078,P = 0.043], longer mechanical ventilation duration [days: 4 (0, 17) vs. 1 (0, 6),U = 7 673.000,P = 0.006], longer length of hospital stay before blood was drawn for culture [days: 21 (7, 40) vs. 9 (3, 17),U = 6 441.500,P = 0.006], and higher incidence of pre-admission intravenous use of antibiotics [84.62% (77/91) vs. 66.83% (139/208),χ2 = 9.989,P = 0.002]. Multiple logistic regression analysis showed that advanced age [odd ratio (OR) = 1.032, 95% confidential interval (95%CI) = 1.015-1.050,P = 0.000], cerebrovascular diseases (OR = 2.247, 95%CI = 1.234-4.090,P = 0.008), prolonged mechanical ventilation (OR =1.041, 95%CI = 1.014-1.069,P = 0.003), and recent hospital stay (≥2 days) within 90 days (OR = 1.968, 95%CI =1.079-3.592,P = 0.027) were the independent risk factors for polymicrobial infection. In the polymicrobial infection group, the length of ICU stay [days: 46 (22, 77) vs. 13 (7, 22),U = 3 148.000,P = 0.000] and hospital stay [days:81 (47, 118) vs. 28 (17, 46),U = 3 620.000,P = 0.000] were significantly longer, and the ICU mortality [65.93%(60/91) vs. 43.75% (91/208),χ2 = 12.463,P = 0.000] and hospital mortality [68.13% (62/91) vs. 45.67% (95/208),χ2 = 12.804,P = 0.000] were significantly higher, and on the other hand the 90-day survival rate was significantly lower than that in the monomicrobial infection group (χ2 = 8.513,P = 0.004).Conclusions The most common pathogen of ICU sepsis is gram-positive cocci. Independent risk factors for polymicrobial infections were found to be advanced age, occurrence of cerebrovascular disease, prolonged mechanical ventilation, and recent hospitalization. Polymicrobial infection is associated with longer length of ICU and hospital stay, as well as higher mortality.

Objective To investigate the risk factors and prognosis of blood stream infection in patients of intensive care unit (ICU).Methods Clinical data of all patients with culture-positive sepsis were collected from all ICUs of Guangdong General Hospital from October 12th, 2012 to December 1st, 2014 for retrospective study.Physiological characteristics and laboratory data were compared between patients with blood culture-positive sepsis group and patients without sepsis of control group.Logistic regression analysis was made to identify the risk factors for blood stream infection.Patients with blood culture-positive sepsis group were further divided into survivor and non-survivor groups according to the clinical outcomes.Physiological and laboratory data were compared between two groups.Logistic regression analysis was also performed to identify the risk factors for mortality.Results There were 299 patients with positive blood culture sepsis admitted in the ICUs in two years.Of them, 250 patients infected with Gram positive cocci including staphylococcus haemolyticus, staphylococcus epidermidis, staphylococcus capitis and staphylococcus aureus accounting for the majority.There were 174 patients infected with Gram negative bacilli including acinetobacter baumannii, Escherichia coli and Klebsiella pneumoniaesubsp.pneumoniae accounting for the majority.A univariate analysis demonstrated that there were significant differences in hypertension (P =0.001), diabetes (P =0.01), coronary heart diseases and heart failure (P =0.000), chronic renal insufficiency (P =0.000), prolonged mechanical ventilation (P =0.000), pre-admission intravenous administration of antibiotics (P =0.000), and hypoalbuminemia (P =0.008) between culture positive group and control group.A logistic regression analysis demonstrated that diabetes [OR =2.158, 95％ CI (1.230, 3.787), P =0.007], chronic renal insufficiency [OR =13.410, 95％ CI (1.715, 104.879), P =0.013], pre-admission intravenous administration of antibiotics [OR =8.375, 95％ CI (5.267, 13.317), P=0.000] were independent risk factors for bloodstream infections in ICU.In patients with positive blood culture, the non-survivor group had patients with higher advance of old age, higher rate of hypertension, coronary heart diseases or congestive heart failure, tumor and chronic renal insufficiency, prolonged mechanical ventilation and higher incidence of surgery and pre-admission intravenous administration of antibiotics compared with the survivor group.The advance of old age [OR =1.023, 95％ CI (1.008-1.037), P =0.002], prolonged mechanical ventilation [OR =1.055, 95％ CI (1.024, 1.088), P =0.000] and hypoalbuminemia [OR =0.933, 95％ CI (0.898, 0.971), P =0.001] were independently associated with mortality of bloodstream infection in ICU.Conclusions Diabetes, chronic renal insufficiency and pre-admission intravenous administration of antibiotics were associated with the development of blood stream infection in ICU.The advance of old age, prolonged mechanical ventilation and hypoalbuminemia were independent risk factors for mortality in patients with culture-positive sepsis in ICU.

Objective To investigate microbial characteristics and predisposing factors in gram-negtive bacteria blood stream infection. Methods A descriptive retrospective study was conducted. Patients diagnosed as sepsis with blood culture of G- bacilli and without sepsis were enrolled. The patients were all admitted to ICUs of Guangdong General Hospital from October, 2012 to December, 2014. The clinical characteristics and outcomes were compared. Multiple logistic regression was used to analyse the predisposing factors for sepsis of G- bacilli. Results A total of 148 patients suffered from sepsis of G-bacilli including Acinetobacter baumannii, Escherichia coli and Klebsiella pneumoniae were enrolled. Single-factor analysis showed that patients with sepsis of G- bacilli infection had older ages, higher incidence of coronary heart diseases or congestive heart failure, cerebrovascular diseases or chronic renal insufficiency, hypertension, also higher incidence of longer length of hospital stay before blood was drawn for culture, and higher incidence using of vasoactive agents and pre-admission intravenous antibiotics and lower plasma albumin level (P < 0.05). Conclusions Coronary heart disease or congestive heart failure, chronic renal insufficiency and pre-admission intravenous antibiotics were independent predisposing factors for sepsis of G-bacilli.

Objective:To summarize the characteristics of nailfold capillaroscopy (NC) in patients with systemic lupus erythematosus (SLE) and explore its clinical significance.Methods:NC examination was performed in 162 SLE patients. The clinical data of SLE patients was collected. Tianniu NC scoring standard was used. The t test was applied to analyze the measurement data, the χ2 test was applied to analyze the counting data. the Pearson or Spearman test was used to evaluate the correlative factors of NC in patients with SLE. Results:NC abnormalities were seen in 87.7%(142/162) of SLE patients, and the incidence of mild, moderate and severe abnormalities was 29.0%(47 cases), 45.1%(73 cases) and 13.6%(22 cases) respectively. The most common NC abnormal manifestation in SLE patients was decreased blood flow velocity (86.4%). In patients with moderate to severe NC abnormalities, the proportions of patients with Raynaud's phenomenon (37.9% vs 23.9%, χ2=2.955, P=0.043) and interstitial lung disease (8.0% vs 0, χ2=5.213, P=0.023), and the level of D-Dimer [(1 992±2 279) μg/L vs (1 248±1 721) μg/L, t=-1.624, P=0.013] were significantly higher than those in the groups with normal/mild NC abnormalities. Correlation analysis demonstrated that Raynaud's phenomena, interstitial lung disease, pulmonary hypertension and D-Dimer were positively correlated with the NC abnormality. Conclusion:NC abnormalities are common in SLE patients. Decreased blood flow velocity is the most frequent manifestation. SLE patients with moderate to severe NC abnormalities should be actively screened for pulmonary hypertension and interstitial lung disease.

Objective:To explore the clinical characteristics of adult-onset non-radiographic axial spondyloarthritis (nr-axSpA) in different genders.Methods:A total of 662 patients with adult-onset nr-axSpA (age at disease onset ≥16 years) who visited the Rheumatology Department of the First Affiliated Hospital of Shantou University Medical College from 1999 to 2020 were included in the study. Comparisons of baseline demographic and clinical characteristics between different genders were performed.Results:Overall, the male-to-female ratio was 1.17∶1, and the prevalence of human leukocyte antigen-B27 (HLA-B27) positivity was 71.8%(475/662). The median baseline disease duration and age at diagnosis was 1.6 (0.5, 4.0) years and 25.0 (21.0, 33.0) years respectively. The males had a significantly earlier age at disease onset and diagnosis [21.0 (18.0, 28.0) vs 25.0 (21.0, 30.0), Z=5.63, P<0.001; 24.0 (19.0, 32.0) vs 27.0 (23.0, 34.5), Z=4.90, P<0.001, respectively] than females. HLA-B27 positivity was more frequent in males than in females [78.4% (280/357) vs 63.9%(195/305), χ2=17.06, P<0.001]. The prevalence of inflammatory back pain (IBP), morning stiffness, nocturnal pain, enthesitis, hip and groin pain were higher in males, whereas females showed a higher prevalence of small joint involvement of the hands. At baseline, males had higher median ankylosing spondylitis disease activity score (ASDAS)-C-reaction protein (CRP) [3.0(2.3, 3.8) vs 2.4(2.0, 3.0), Z=5.59, P<0.001] and a greater prevalence of high disease activity ASDAS-CRP>2.1 [81.9%(185/227) vs 67.9%(133/195), χ2=11.08, P=0.001] than females. The proportions of male patients with elevated CRP levels and erythrocyte sedimentation rate (ESR) were also higher than those of female patients [49.0%(175/357) vs 27.9%(85/305), χ2=30.85, P<0.001; 49.3%(176/357) vs 33.4%(102/305), χ2=16.98, P<0.001, respectively]. Conclusion:The adult-onset nr-axSpA in China is characterized by a comparable sex ratio. Males have an earlier age at disease onset and are higher HLA-B27 positivity with higher prevalence of IBP, enthesitis, hip and groin pain, as well as high disease activity.

Systemic lupus erythematosus (SLE) complicated with acquired hemophilia A (AHA) is a rare condition with frequently delayed diagnosis and a high mortality rate, so it is necessary to strengthen the understanding of this disease. In this study, the characteristics and treatment in 1 case of SLE complicated by AHA is reported and analyzed, and a literature review is conducted. The patient was a 29-year-old young female with a 10-year history of SLE, the main clinical manifestation was severe abdominal bleeding. Laboratory tests revealed that the activated partial thromboplastin time (APTT) was notably prolonged (118.20 s), and the coagulation factor VIII activity (FVIII꞉C) was extremely decreased (0.20%) with high-titer of factor VIII (FVIII) inhibitor (31.2 BU/mL). After treating with high-dose glucocorticoid, immunoglobulin, cyclophosphamide, rituximab, blood transfusion, and intravenous infusion of human coagulation FVIII, the coagulation function and coagulation FVIII꞉C were improved, and FVIII inhibitor was negative without serious adverse reactions. During the next 5-year follow-up, the patient's condition was stable and no bleeding occurred. In the case of coagulation dysfunction in SLE, especially with isolated APTT prolongation, AHA should be screened. When the therapeutic effects of glucocorticoid combined with immunosuppressants are not desirable, rituximab could be introduced.
Female ; Humans ; Adult ; Hemophilia A/therapy* ; Rituximab ; Glucocorticoids ; Factor VIII ; Lupus Erythematosus, Systemic/complications* ; Hemorrhage/complications*

CRISPR-Associated Protein 9 ; Gene Editing ; CRISPR-Cas Systems/genetics*

COVID-19 ; Coronavirus Nucleocapsid Proteins ; Humans ; Nucleocapsid Proteins ; Phosphoproteins ; SARS-CoV-2