Objective To observe the clinical effect and safety of umbilical cord blood mesenchymal stem cell(UCB-MSC) transplantation in the treatment of diabetic foot. Methods UCB-MSC suspension (cell concentration (3 -7) × 107/L,0.3 -0.5 ml per point) was injected into multiple spots on affected lower limb with a 3 cm × 3 cm istance among each point. Demixing injection could be performed in regions with multilayer muscles. Clinical symptoms and related index were routinely observed from the first day to three months after operation. Results After three months of UCB-MSC transplantation, pain of patients was relieved, skin temperature increased, intermittent claudication ameliorated, ulcer healed, ankle-brachial index and transcutaneous partial pressure of oxygen increased. The lower extremity lesions showed an abundant collateral vessel formation after the treatment in 2 patients by angiography. Both patients had no severe complications and adverse reactions, none underwent amputation. Conclusions Umbilical cord blood mesenchymal stem cell transplantation is a safe and effective treatment of diabetic lower limb ischemic disease, which can exempt the patient from amputation and improve their quality of life.

Objective To investigate the effects of allogenic intra-bone marrow bone marrow transplantation (IBM-BMT) on re-establishing hematopoiesis in mice. Methods Bone marrow mononuclear cells (BMNCs) from BALB/ c mice were transplanted into the C57BL/6 mice treated with a lethal dose of ~(60)Coγ-ray radiation through intra-bone marrow injection or intravenous injection. Sixty of the C57BL/6 mice were randomly divided into three groups as higher dose intra-bone marrow injection group (IBM1 group), lower dose intra-bone marrow injection group (IBM2 group) and intravenous injection group (IV group). The nucleated cell numbers of whole bone marrow from the tibia of each recipient mouse were counted respectively at the day 1, day 3, day 6 and day 9 after the transplantation. The donor-derived total nucleated cells and myeloid cells were quantified by flow cytometry. Results At 6th day after transplantation, more total bone marrow nucleated cells, total donor-derived nucleated cells and donor-derived myeloid cells in the tibia of injected side in both IBM1 group and IBM2 group were found than that in IV group (P<0.05 or P<0.01). Conclusion Compared with traditional bone marrow transplantation (IV-BMT),IBM-BMT improves the bone marrow hematopoiesis in the early hematopoietic re-establishing stage in allogenic bone marrow transplantation.

To investigate whether human umbilical cord plasma (HUP) can be used to culture human cord blood mesenchymal stem cells (HUCMSCs), we collected 20 surplus HUP. After being treated with salting out and diasysis, the HUP were used to culture HUCMSCs as 10% volume, and compared with fetal bovine serum (FBS). Morphological characteristics, growth curve and reproductive activity of HUCMSCs cells were observed. The concentration of bFGF and noggin secreted by HUCMSCs cultured with HUP and FBS medium were detected by ELISA. It was found that compared to FBS, the morphology, reproductive activity and characteristic of HUCMSCs cell cultured with HUP were not distinctively different from FBS. The concentration of bFGF in HUP group was significantly higher than that of FBS group, and the concentration of noggin was also different in the two groups. So we concluded that HUP could be used to culture HUCMSCs for a long-time, and the HUP mediumcoild could be more suitable for the culture of human embryonic stem cell (hESC).
Animals ; Cattle ; Cell Culture Techniques ; Cells, Cultured ; Culture Media ; chemistry ; Fetal Blood ; chemistry ; Humans ; Mesenchymal Stromal Cells ; cytology ; Plasma ; chemistry ; Serum ; chemistry

Objective The objective of this study was to investigate the effect of acidic conditions on epithelial mesenchymal transition(EMT) of HepG2 cells.Methods HepG2 cells were cultured under acidic and alkaline conditions to observe the difference of cell morphology.Wound healing assays were performed to detect the migration ability of the two groups.Matrigel invasion assays were used to detect the invasive ability of the cells.The expression of EMT-related genes at mRNA level was detected by RT-PCR.The expression level of EMT-related gene protein was detected by Western blot.Results The morphology of HepG2 cells was changed from epithelium to interstitial type under acid conditions.The migration ability of HepG2 cells under acidic condition was stronger than that of alkaline conditions.The number of HepG2 cells crossing Matrigel was higher than that of alkaline condition.The mRNA expression of Vimentin,Slug,Snail and Zeb1 related to EMT and the protein expression of Vimentin and MMP9 related to EMT in HepG2 cells were significantly higher than those under alkaline condition.Conclusion The acid conditions can induce the occurrence of EMT in HepG2 cells.

Objective To evaluate the differences among five representative and useful Doppler parameters in the diagnosis of the three common types of renal artery stenosis (RAS). Methods Five Doppler parameters including renal peak systolic velocity (RPSV), renal-aortic ratio (RAR), renal-interlobar ratio (RIR),acceleration time (AT),and resistant index (RI) were measured in 221 patients before renal arteriography. Differences between the groups of patients with various clinical causes of RAS were analyzed by Chi-Squared test. One-way ANOVA or t test were used to compare the means between different groups.The optimal cutoff value was determined with the maximum sum of sensitivity and specificity. Results Of the 442 renal arteries (main and accessory renal arteries) demonstrated at arteriography,214 were normal or stenosed less than 50 %, 204 stenoses 50% - 99 %, and 24 occlusions. RIR, RAR and RI were significantly different between the atherosclerotic and non-atherosclerotic RAS groups (P ＜0.05), while RPSV and AT were not. The optimal cutoff values of RAR,RIR and RI for detecting RAS between the atherosclerotic and non-atherosclerotic groups were much different (2.5 versus 1.9, 5. 1 versus 6.5, 0.57 versus 0.50,respectively) ,but those of RPSV and AT were similar or the same (170 cm/s versus 200 cm/s,51 ms versus 51 ms,respectively). Conclusions In the case of RAS (diameter reduction≥50%),it is advised to establish separate cutoff values of RAR, RIR and RI according to atherosclerotic and non-atherosclerotic RAS, but the same cutoff value of RPSV and AT can be applied. RIR is a good Doppler parameter in the diagnosis of RAS,especially atherosclerotic and fibromuscular dysplasia RAS.

Objective:To investigate the incidence of thyroid dysfunction in patients with chronic kidney disease and analyze the influencing factors.Methods:One hundred and ninety-eight patients with chronic kidney disease who were treated in Chronic Disease Management Department of the First Affiliated Hospital of Hunan University of Traditional Chinese Medicine from Apr. 2021 to Apr. 2022 were selected, including 71 patients with abnormal thyroid function and 127 patients with normal thyroid function. The differences in TT3, FT3, TT4, FT4, and TSH between patients with abnormal thyroid function and those with normal thyroid function were analyzed. At the same time, the abnormal thyroid function of patients with different clinical characteristics and the influencing factors were analyzed. The intergroup differences were analyzed using t-test or χ2-test, and the influencing factors were analyzed using logistic regression analysis. Results:In one hundred and ninety-eight patients with chronic kidney disease, thyroid function abnormality occurred in 71 patients (35.86%), including two or more abnormal thyroid function indicators in 35 patients (49.30%). The total triiodothyronine (TT3), free triiodothyronine (FT3), total thyroxine (TT4) and free thyroxine (FT4) in patients with abnormal thyroid function were (1.02 ± 0.29) nmol/mL, (3.03 ± 0.88) pmol/L, (77.93 ± 20.02) nmol/mL and (11.02 ± 1.95) pmol/L respectively, which was significantly lower in patients with normal thyroid function (1.32±0.25) nmol/mL, (4.20±0.92) pmol/L, (93.30±19.28) nmol/mL and (13.54±1.82) pmol/ ( P<0.05), while thyroid stimulating hormone (TSH) was (3.14 ± 0.96) mIU/L, which was significantly higher than that in patients with normal thyroid function (1.84±0.89) mIU/L ( P<0.05). The incidence of thyroid dysfunction in female patients was 50.59% (43/85), It was significantly higher than 24.78% (28/113) of male patients (P <0.05) ; The incidence of thyroid dysfunction in patients aged 60 years was 49.55% (55/111), It was significantly higher than 18.39% (16/87) of the patients aged <60 years ( P<0.05) ; The incidence of thyroid dysfunction in patients with 1-year duration of disease was 71.43% (30/42), It was significantly higher than 25.28% (41/156) of patients with a course of disease <1 year ( P<0.05) ; The incidence of thyroid dysfunction in patients with clinical stage G 4 to 5 was 50.62% (41/81), It was significantly higher than 25.64% (30/117) of patients in G1~3 stages ( P<0.05) ; The incidence of thyroid dysfunction in patients with diabetes was 74.36% (29/39), This was significantly higher than 26.42% (42/159) in patients without diabetes mellitus ( P<0.05). Logistic regression analysis showed that gender, age, course of disease and clinical stage were the influencing factors of thyroid dysfunction in patients with chronic kidney disease ( P<0.05) . Conclusion:A high proportion of patients with chronic kidney disease have abnormal thyroid function, which is affected by the patient's sex, age, course of disease and clinical stage,clinical attention should be paid to targeted intervention to prevent the incidence of thyroid dysfunction in chronic kidney disease population.

Objective:This study evaluates the performance of chemiluminescence assay, which is designed to detect Hepatitis D Virus (HDV) Immunoglobulin G (IgG) antibodies.Methods:A comparative analysis was conducted among chemiluminescence anti-HDV IgG reagent, the magnetic particle-based domestic reagent A and domestic reagent B, and the Robo Gene HDV RNA kit, using 1909 HBsAg-positive plasma samples. This comparison aimed to delineate clinical specificity and detection accuracy. The anti-HDV IgG reagent precision was assessed at three different concentration levels following the Clinical Laboratory Standards Institute EP5-A2 guidelines. The specificity of the assay was validated using 200 HAV IgM positive, 545 HBsAg-positive but anti-HDV IgG-negative, 350 anti HCV positive plasma samples and 200 healthy human blood samples. Additionally, a concordance study was conducted with 545 HBsAg-positive and 37 anti-HDV IgG-positive plasma samples, comparing the anti-HDV IgG reagent against reagent A.Results:1 909 HBsAg-positive plasma samples were tested using 3 anti HDV IgG reagent and 1 HDV RNA reagent, 19 samples were identified as anti-HDV IgG-positive. The anti-HDV IgG demonstrated superior accuracy and specificity. The assay exhibited excellent precision, with intra-assay coefficient of variation (CV) values ranging from 1.57% to 4.30%, and inter-assay CV values between 1.71% and 4.67% for detecting samples at high, medium, and low concentration levels. Concordance with Reagent A showed consistent results in both positive and negative detections.Conclusion:In this study, the anti-HDV IgG reagent (chemiluminescence method) displayed outstanding specificity in detecting clinical samples and exhibited a high conformity rate with commercialized reagents, making it potentially suitable for screening anti-HDV IgG in HBsAg-positive samples.

Objective:This study aims to evaluate the quality and explore the preliminary clinical applications of a domestically developed hepatitis D virus nucleic acid quantification reagent (abbreviated as"domestic HDV RNA reagent").Methods:The sensitivity and accuracy of the reagent were evaluated in accordance with the WHO HDV RNA international standard, employing the Bio-Rad CFX Opus 96 real-time fluorescence quantitative PCR analysis system. Serial dilutions of pseudo-viruses or cell culture-derived virus were used to determine the linear range of the domestic HDV RNA reagent. Specificity was assessed using positive samples of HAV, HBV, HCV infection, and HEV national reference materials. Precision was evaluated with samples at both high and low concentrations. In a comparative analysis, 30 HDV IgG positive samples were tested using both the domestic HDV RNA reagent and the RoboGene HDV RNA kit based on the ABI 7500 FAST DX system. The Pearson correlation coefficient (r) was used to examine the correlation between the two reagents.Results:The domestic HDV RNA reagent demonstrated a high sensitivity of up to 6 IU/ml, consistent with that of the comparator reagent. The calibration curve for WHO HDV RNA standards had a slope of -3.286, with an amplification efficiency of 101.6%. The linear detection range spanned from 10 to 10 8 IU/ml for eight HDV genotypes. The domestic HDV RNA reagent exhibited exceptional specificity, without cross-reactivity observed with HAV, HBV, HCV, or HEV. Accuracy assessments at five concentration levels met the required standards, with intra-assay precision coefficient of variation ( CV) ranging from 1.20% to 4.20%, and inter-assay precision CV from 1.20% to 7.90%. The detection results for HDV IgG positive samples were highly correlated with the comparator reagent ( r=0.984, P<0.001), achieving a diagnostic accuracy of 100% compared to sequencing results. Conclusion:In this study, the domestic HDV RNA reagent possesses excellent specificity, accuracy, precision, and a broad linear range, attaining a sensitivity level on par with international reagents of the same type.

For a long time, macrolides have been the first choice for the antibacterial treatment for pertussis.However, in the past decade, resistance to macrolide antimicrobials has been common in clinically isolated Bordetella pertussis in China, which is in contradiction with the recommended macrolide treatment.Therefore, Trimethoprim-Sulfamethoxazole (TMP-SMZ) is suggested as the first choice for antibacterial treatment for pertussis in China, with a dosage determined according to age and body weight, lasting 14 days.If TMP-SMZ cannot be used, full-dose and full-course β-lactam antimicrobials may be used, of which the effects should be assessed carefully.The impact of other antibacterial drugs, such as quinolones and tetracyclines, on the elimination of Bordetella pertussis should also be evaluated as soon as possible to treat adult pertussis and potential cases caused by drug-resistant bacteria in future.

Objective:To determine the erythromycin resistance of Bordetella pertussis isolates and their ptxP1 and ptxP3 phenotypic composition and compare clinical manifestations of children with pertussis caused by the two types of strains. Methods:This was a cross-sectional study, the pertussis cases diagnosed using bacterial culture from January 2019 to December 2022 in Beijing Children′s Hospital and the First People′s Hospital of Wuhu were collected.Any suspected Bordetella pertussis colonies were identified by the slide agglutination test.The susceptibility of isolates to erythromycin was detected by the E-test and K-B test.The ptxP gene was amplified by polymerase chain reaction and sequenced to determine its genotype. t-test, Mann-Whitney U-test, Chi-square test and Fisher′s exact test were use to statistical analysis. Results:A total of 192 strains of Bordetella pertussis were identified, including 188 (97.9%) erythromycin-resistant strains.Among the 188 strains, 30.3%(57/188) belonged to the ptxP1 genotype and 69.7%(131/188) belonged to the ptxP3 genotype.In children aged below 1 year old, the incidence of paroxysmal cough caused by infection with the ptxP3 strain was higher than that with the ptxP1 strain (57.1% vs.29.4%, P<0.05), and children infected with the ptxP3 strain were more likely to develop apnea or asphyxia (23.8% vs.17.6%), post-tussive vomiting (44.4% vs.32.4%), whooping cough (72.0% vs.50.0%) and pneumonia or bronchitis (85.7% vs.73.5%) compared to those infected with the ptxP1 strain, but the differences were not statistically significant(all P>0.05). In children aged 1 year old and above, the white blood cell count of children infected with the ptxP1 strain was higher than that of infections with the ptxP3 strain [13.5(9.9, 24.5)×10 9/L, 10.3 (7.0, 16.4)×10 9/L, P<0.05], and children infected with the ptxP1 strain were more likely to contract other pathogen infections than those infected with the ptxP3 strain (17.4% vs.4.4%, P>0.05). Conclusions:ptxP3 erythromycin-resistant Bordetella pertussis has become the main pathogen of pertussis.Infants with pertussis caused by the ptxP3 erythromycin-resistant strain show more significant manifestations and a higher possibility of severe symptoms than those infected with the ptxP1 erythromycin-resistant strain.