The coagulation VIII factor (FVIII) contains eight pairs of disulfide bonds, which are involved in maintaining its structure and function. It has been demonstrated that the disulfide bond between Cys¹⁸⁹⁹/Cys¹⁹⁰³ of the A3 domain in the light chain impedes secretion. In our previous work, an engineered inter-chain disulfide in the B domain-deleted FVIII (BDD-FVIII) promoted heterodimer assembly and secretion of separately expressed heavy and light chains. In this study, we constructed two BDD-FVIII variants, one of which contains an engineered inter-chain disulfide bond (F8C) between Met⁶⁶² > Cys and Asp¹⁸²⁸ > Cys mutations and another contains an endogenous A3 domain with a disrupted disulfide bond from F8C (F8CG) by replacement of Cys¹⁸⁹⁹ and Cys¹⁹⁰³ with Gly in F8C. We explored their function and secretion. By transducing F8C and F8CG into HEK293 and COS-7 cells, the formation of disulfide bonds and the secretion and coagulation activity of the two variants in the culture media and their binding affinity for von Willebrand factor (vWF) could be observed. The results show that variants F8C and F8CG are mainly the disulfide bonded heavy and light chain dimer, while the wild type BDD-FVIII (F8) is dominated by the easily dissociated heavy and light chain dimer. The secretion and activity of F8C was significantly higher than that of F8, while the secretion and activity of F8CG was significantly higher than that of F8C. The vWF binding of the two variants is similar to F8. This indicates that the BDD-FVIII variant F8CG may be attractive molecule for protein replacement and as a transgene in gene-therapy strategies. These findings are encouraging for future studies targeting disulfide bond elimination for further enhancement of FVIII secretion.

OBJECTIVE: To investigate the value of red blood cell distribution width (RDW) and fibrinogen (Fib) level for the evaluation of therapeutic efficacy and prognosis in patients with diffuse large B-cell lymphoma (DLBCL). METHODS: The relationship between RDW/Fib at initial diagnosis and efficacy and the clinical outcome was retro-spectively analyzed based on the study of 105 patients with DLBCL. The patients were divided into two groups: low RDW group (≤15%) and high RDW group (>15%), low Fib group (Fib≤4 g/L) and high Fib group (Fib>4 g/L) according to the normal values of RDW and Fib. Therapeutic efficacy, overall survival (OS) time and progression free survival (PFS) time were compared between two groups. The correlation between each factors and efficacy, prognosis was analyzed by univariate and multivariate regression. RESULTS: The therapeutic efficacy (P<0.001), OS time(P=0.004), and PFS time(P=0.007) were poorer in the high RDW group as compared with the low RDW group. The efficacy (P=0.015) and PFS time(P=0.04) were poorer in the high Fib group as compared with the low Fib group. Multivariate analysis showed that high RDW was the independent risk factor for efficacy of DLBCL patients (OR=3.394, 95% CI 1.093-10.539, P=0.035). CONCLUSION High RDW and high Fib associate with poor efficacy in DLBCL patients.

OBJECTIVE: To investigate the efficacy and safety of arsenic trioxide combined with ATRA and chemo- therapy for treatment of relapsed acute promyelocytic leukemia (APL) patients. METHODS: The clinic data of 25 patients with relapse APL treated in our hospital from 1996 to 2013 were collected and analyzed. Among the 25 patients, 15 patients suffered first-time hematological relapse (HR), and the other 10 patients showed first-time molecular relapse (MR). The patients with first-time replase were treated with ATO+ATRA+Anthracycline re-induction chemotherapy. The clinical features, complete remission (CR) rate, overall survival (OS), disease-free survival (DFS) and adverse events after re-induction therapy were analyzed. RESULTS: Fourteen of 15 hematological relapsed patients achieved the second-time hematological complete remission (CR2) after re-induction therapy except one patient died of bleeding complication during the re-induction. 8 of 14 patient showed molecular complete remission (CRm) after two cycles of therapy with this regimen. Totally, eleven out of the 14 HR patients were alive without disease till the last follow-up, and 3 of the 14 HR patients died because of bleeding complications. All of the 10 molecular relapsed patients received the second CRm after treated by the regimen. Among these 10 patients, 6 patients suffered only once relapse and continued with the molecular CR2 status, and for the other 4 patients with more than two-relapses, only 1 survived untill 89.3 months after achieved second-time CRm, and other 3 patients died because of bleeding complications. CONCLUSION For relapsed APL patients, the treatment with ATO+ATRA+chemotherapy regimen after relapse still shows encouraging efficacy, no matter whether or not the application of ATO in the previous regimens. In addition, patients with more than two molecular relapses show a poor prognosis.

OBJECTIVE: To study the changes in the serum levels of Chemerin and Omentin-1 in children with Kawasaki disease (KD) in the acute stage after intravenous immunoglobulin (IVIG) treatment and related clinical significance. METHODS: A total of 60 children who were diagnosed with KD from January 2015 to April 2019 were enrolled as subjects. Forty healthy children and 40 children with acute infectious diseases were enrolled as the healthy control group and the infection control group respectively. According to the sensitivity to IVIG treatment, the children with KD were divided into an IVIG sensitive group with 51 children and a non-IVIG sensitive group with 9 children. According to the presence or absence of coronary artery lesion, the children with KD were divided into a CAL group with 13 children and a non-CAL group with 47 children. ELISA was used to measure the serum levels of Omentin-1 and Chemerin before and after the treatment. RESULTS: The children with KD had significantly higher serum levels of Chemerin and Omentin-1 than the healthy control and infection control groups before treatment (P<0.05). After 48 hours of treatment, the IVIG sensitive group had a significant reduction in the serum level of Chemerin (P<0.05), while there was no significant change in the serum level of Omentin-1 after treatment (P>0.05). Before treatment, the non-IVIG sensitive group had a significantly higher serum level of Chemerin than the IVIG sensitive group (P<0.05), and the CAL group had a significantly higher serum level of Chemerin than the non-CAL group, while there was no significant difference in the serum level of Omentin-1 between the IVIG sensitive and non-IVIG sensitive groups, as well as between the CAL and non-CAL groups (P>0.05). CONCLUSIONS High serum levels of Chemerin and Omentin-1 may play an important role in the development and progression of KD. Chemerin may be involved in the development of CAL in children with KD. The serum level of Chemerin may be used as a new index for predicting the sensitivity to IVIG treatment.
Adipokines ; Chemokines ; Child ; Coronary Artery Disease ; Humans ; Immunoglobulins, Intravenous ; Mucocutaneous Lymph Node Syndrome

OBJECTIVE: To evaluate the efficacy and safety of C-reactive protein (CRP)-guided antibiotic treatment strategy for neonates with suspected early-onset sepsis (EOS). METHODS: A total of 428 neonates, with a gestational age of >35 weeks, who were admitted to the Children's Hospital of Chongqing Medical University from February to July, 2019 and were suspected of EOS were enrolled as the observation group. The effect of antibiotic treatment was prospectively observed, and if clinical symptoms were improved and CRP was <10 mg/L in two consecutive tests, discontinuation of antibiotics was considered. A total of 328 neonates (gestational age of >35 weeks) who were admitted to this hospital from February to July, 2018 and were suspected of EOS were enrolled as the control group, and the use of antibiotics was analyzed retrospectively. The two groups were compared in terms of duration of antibiotic treatment, length of hospital stay, incidence rate of repeated infection and clinical outcome. RESULTS: Compared with the control group, the observation group had significantly shorter duration of antibiotic treatment and length of hospital stay (P<0.05). There were no significant differences in the incidence rate of repeated infection and clinical outcome between the two groups (P>0.05). CONCLUSIONS For neonates with a gestational age of >35 weeks and a suspected diagnosis of EOS, CRP-guided antibiotic treatment strategy can shorten duration of antibiotic treatment and length of hospital stay and does not increase the incidence rate of repeated infection. Therefore, it holds promise for clinical application.
Anti-Bacterial Agents ; therapeutic use ; C-Reactive Protein ; Gestational Age ; Humans ; Infant, Newborn ; Retrospective Studies ; Sepsis ; drug therapy

The problems of the syndrome differentiation of migraine in acupuncture treatment were collected, e.g. inconsistency of syndrome differentiation, unclear staging of syndrome differentiation, lack of standardization in comparison between syndrome differentiation and non-differentiation, insufficient research on the factors of syndrome differentiation. In view of the exiting problems, focusing on two aspects of migraine, namely syndrome differentiation and acupuncture effect, the clinical treatment and research are conducted in migraine treated with acupuncture based on syndrome differentiation. It is believed that the comprehensive observation of the relationship between syndrome differentiation and acupuncture effect, as well as the analysis of the relevant factors of syndrome differentiation of migraine should be the focus in future research.
Acupuncture Therapy ; Humans ; Migraine Disorders ; therapy ; Treatment Outcome

200 µm, a circumference > 60 degrees, and a cap thickness < 450 µm. The percentage distributions of CT pixel attenuation ≤ 20, 30, 40, and 50 HU were calculated using quantitative histogram analysis.RESULTS: A total of 271 transverse sections were co-registered between CCTA and pathological analysis. Overall, 26 lipid cores and 16 fibrous plaques were identified by pathological analysis. There was no significant difference in median CT attenuation between the lipid and fibrous plaques (51 HU [interquartile range, 46–63] vs. 57 HU [interquartile range, 50–64], p = 0.659). The median percentage of CT pixel attenuation ≤ 30 HU accounted for 11% (5–17) of lipid-core plaques and 0% (0–2) of fibrous plaques (p < 0.001). The sensitivity and specificity of the method for diagnosing lipid plaques by the average CT pixel attenuation ≤ 30 HU were 80.8% and 87.5%, respectively. The area under the receiver operator characteristics curve was 0.898 (95% confidence interval: 0.765–0.970; 3.0% was the best cut-off value). The diagnostic performance was significantly higher than those of the average pixel CT attenuation percentages ≤ 20, 40, and 50 HU and the mean CT attenuation (p < 0.05).CONCLUSION: In in vivo conditions, with the pathological lipid core as the gold standard, quantification of the percentage of average CT pixel attenuation ≤ 30 HU in the histogram can be useful for accurate identification of lipid plaques.]]>
Angiography ; Coronary Disease ; Coronary Vessels ; Diagnosis ; Heart Transplantation ; Humans ; Methods ; Sensitivity and Specificity ; Spectrum Analysis

Objective: To recover broad-neutralizing monoclonal antibodies (BnAbs) from avian influenza A (H5N1) virus infection cases and investigate their genetic and functional features. Methods: We screened the Abs repertoires of expanded B cells circulating in the peripheral blood of H5N1 patients. The genetic basis, biological functions, and epitopes of the obtained BnAbs were assessed and modeled. Results: Two BnAbs, 2-12D5, and 3-37G7.1, were respectively obtained from two human H5N1 cases on days 12 and 21 after disease onset. Both Abs demonstrated cross-neutralizing and Ab-dependent cellular cytotoxicity (ADCC) activity. Albeit derived from distinct Ab lineages, , V 1-69-D2-15-J 4 (2-12D5) and V 1-2-D3-9-J 5 (3-32G7.1), the BnAbs were directed toward CR6261-like epitopes in the HA stem, and HA I45 in the hydrophobic pocket was the critical residue for their binding. Signature motifs for binding with the HA stem, namely, IFY in V 1-69-encoded Abs and LXYFXW in D3-9-encoded Abs, were also observed in 2-12D5 and 3-32G7.1, respectively. Conclusions Cross-reactive B cells of different germline origins could be activated and re-circulated by avian influenza virus. The HA stem epitopes targeted by the BnAbs, and the two Ab-encoding genes usage implied the VH1-69 and D3-9 are the ideal candidates triggered by influenza virus for vaccine development.

Objective To analyze the resistance of influenza virus to neuraminidase inhibitors (NAI) in Hebei province during 2018-2019. Methods Virus were collected from the Hebei Influenza Surveillance Network during 2018-2019. A total of 36 confirmed influenza viruses (with 25 H1pdm09 and 11 H3N2) were selected to test resistance to oseltamivir and zanamivi with fluorescence (FL). Results All 36 influenza viruses tested were sensitive to oseltamivir and zanamivir. The median half maximal inhibitory concentration (IC50) for oseltamivir of H1pdm09 and H3N2 were of 0.50 nM (range 0.07-1.14 nM) and 0.25 nM (range 0.09-0.69 nM) respectively, while 0.29 nM (range 0.09-0.85 nM) and 0.87(range 0.17-1.81 nM) for zanamivir, all were within 10 fold IC50 of the reference virus (corresponding type). Conclusion All the tested influenza strains isolated in Hebei province during 2018-2019 were sensitive to NAI.

Objective: To analyze the dynamic changes in the expression and function of peripheral type Ⅱ innate lymphoid cell (ILC2) subpopulation and the activity of signal transducers and activators of transcription (STAT6) in children with hay fever during pollen season. Methods: A total of 10 patients with hay fever, 10 patients with house dust mite (HDM)-sensitized asthma and 12 healthy controls (HC) were enrolled in this study. Changes in peripheral ILC2 and the intracellular expression of Th2-related cytokines were detected by flow cytometry during and outside the pollen season. Peripheral Lin^- cell population was isolated from each group and cultured with the presence of IL-25 or IL-33 for 7 d. The concentrations of IL-5 and IL-13 in culture supernatants were measured by ELISA. Expression of phospho-STAT6 at protein level was quantified by Western blot. Results: Within the pollen season, the percentage of peripheral ILC2 cells was significantly higher in children with hay fever [(23.09±7.86)%] than in children with HDM-sensitized asthma [(6.84±3.85)%, P<0.05] and healthy children[(1.69±0.87)%, P<0.05]. In the non-pollen season, the peripheral ILC2 cells in children with hay fever presented a decreasing trend [(11.30±2.45)%], but was still higher than that in HDM-sensitized asthmatics [(3.76±1.96)%, P<0.05] and HC [(1.32±0.91)%, P<0.05] at the same time point. Moreover, peripheral IL-13⁺ ILC2 cells in children with hay fever [(6.94±3.16)% vs(4.17±1.98)%, P<0.05] and in HDM-sensitized asthmatics [(1.89±0.70)% vs(1.44±0.55)%, P<0.05] during the pollen season were significantly higher than those in the non-pollen season. After the in vitro stimulation with IL25 or IL-33, the levels of IL-5 and IL-13 in culture supernatants were both increased in children with hay fever and HDM-sensitized asthmatics, and a synergistic action was observed when IL25 and IL-33 were used in combination. Meanwhile, the protein level of phospho-STAT4 in Lin^- cells was significantly up-regulated in the hay fever group after stimulation with IL25 and IL-33. Conclusions During the pollen season, the abnormal number and function of ILC2 subpopulation in children with hay fever might be another cause of the occurrence of clinical symptoms in a short period of time or acute exacerbation.