ObjectiveThrough improving the potential of vascular endothelial growth factor receptor (VEGFR)-humanized mouse model (hKDR^+/+) with C57BL/6N background to allow the growth of different mouse tumor cell lines, to establish novel tumor-bearing mouse models which can support in vivo tumorigenesis of different mouse tumor cell lines and be used to evaluate drugs targeting VEGFR.MethodsFirstly, a method to evaluate the in vivo activity of antibody targeting VEGFR based on the hKDR^+/+ humanized mouse model was established. Recombinant activating gene 1 (Rag1) knockout mice (Rag1^-/-) were established using CRISPR/Cas9 technology. Then these Rag1^-/- mice were crossed with hKDR^+/+ mice to get a double gene modified homozygous hKDR^+/+/Rag1^-/- mouse model by screening. Finally, tumor cell lines derived from different mouse strains were tested on the double gene-modified mouse model to compare the differences in tumor growth. ResultsAntibodies designed for VEGFR showed significant anti-tumor activity in hKDR^+/+ mice, which significantly reduced tumor volume and weight compared with the PBS group (P<0.01, P<0.05). The number of B cells and T cells in the peripheral blood of Rag1^-/- mice and hKDR^+/+/Rag1^-/- mice decreased (P<0.05, P<0.001). Tumors were observed in hKDR^+/+/Rag1^-/-, Rag1^-/-, wild-type, and hKDR^+/+ mice after 7 d of inoculation of MC38 cells derived from C57BL/6 mice. Tumors were only observed in groups of hKDR^+/+/Rag1^-/- and Rag1^-/- mice, but not in the wild-type and hKDR^+/+ mice after 10 d of inoculation with CT26 cells derived from BALB/c mice. After 3 weeks of inoculation, the tumor volume of hKDR^+/+/Rag1^-/- mice was significantly larger than that of Rag1^-/- mice (P<0.01). ConclusionRag1 knockout mice were obtained and a novel hKDR^+/+/Rag1^-/- double genes modified mouse model was further screened. The tumor cell lines from different mouse strain origins were more prone to growth in mice with Rag1 gene deficiency. The results suggest that the reduced immune response of hKDR^+/+ humanized mice will improve the capacity of supporting the growth of mouse tumor lines in the model. As a result, more tumor-bearing mouse models may be constructed for the evaluation of drugs targeting VEGFR in this way.

Objective:To explore the effectiveness, safety and cost between urinary follicle stimulating hormone (uFSH) and recombinant follicle stimulating hormone (rFSH) in controlled ovarian stimulation (COS) in China.Methods:Data were collected from 16 reproductive centers in China covering oocytes collection time from May 1, 2015 to June 30, 2018. Eligible patients were over 18 years old, adopting COS with uFSH (uFSH group) or rFSH (rFSH group) as start gonadotropins (Gn), and using in vitro fertilization (IVF) and (or) intracytoplasmic sperm injection for fertilisation, excluding frozen embryo recovery cycle. Generalised estimating equation was used to address the violation of independency assumption between cycles due to multiple IVF cycles for one person and clustering nature of cycles carried out within one center. Controlling variables included age, body mass index, anti-Müllerian hormone level, cause of infertility, ovulation protocol, type of fertilisation, number of embryos transferred, number of days of Gn use.Results:Totally 102 061 cycles met eligibility criteria and were included in the analyses. In terms of effectiveness, after controlling relevant unbalanced baseline characteristics, compared with rFSH group, the high oocyte retrieval (>15 oocytes was considered high retrieval) rate of uFSH group significantly decreased in gonadotropin-releasing hormone agonist protocol ( OR=0.642, P<0.01) and in gonadotropin-releasing hormone antagonist protocol ( OR=0.556, P=0.001), but the clinical pregnancy rate per transfer cycle and the live birth rate per transfer cycle significantly increased ( OR=1.179, OR=1.169, both P<0.01) in both agonist and antagonist protocols. For safety, multiple analysis result demonstrated that in the agonist protocol, compared with rFSH group, the incidence of moderate to severe ovarian hyperstimulation syndrome of uFSH group significantly decreased ( OR=0.644, P=0.002). The differences in ectopic pregnancy rate and multiple pregnancy rate between the uFSH and rFSH groups were not significant ( P=0.890, P=0.470) in all patients. In terms of cost, compared with rFSH group, the uFSH group had lower total Gn costs for each patient ( P<0.01). Conclusion:For patients who underwent COS, uFSH has better safety, and economic profiles over rFSH in China.

Objective: To investigate the value of initial CT quantitative analysis of ground-glass opacity (GGO), consolidation, and total lesion volume and its relationship with clinical features for assessing the severity of coronavirus disease 2019 (COVID-19). Materials and Methods: A total of 84 patients with COVID-19 were retrospectively reviewed from January 23, 2020 to February 19, 2020. Patients were divided into two groups: severe group (n = 23) and non-severe group (n = 61). Clinical symptoms, laboratory data, and CT findings on admission were analyzed. CT quantitative parameters, including GGO, consolidation, total lesion score, percentage GGO, and percentage consolidation (both relative to total lesion volume) were calculated. Relationships between the CT findings and laboratory data were estimated. Finally, a discrimination model was established to assess the severity of COVID-19. Results: Patients in the severe group had higher baseline neutrophil percentage, increased high-sensitivity C-reactive protein (hs-CRP) and procalcitonin levels, and lower baseline lymphocyte count and lymphocyte percentage (p < 0.001). The severe group also had higher GGO score (p < 0.001), consolidation score (p < 0.001), total lesion score (p < 0.001), and percentage consolidation (p = 0.002), but had a lower percentage GGO (p = 0.008). These CT quantitative parameters were significantly correlated with laboratory inflammatory marker levels, including neutrophil percentage, lymphocyte count, lymphocyte percentage, hs-CRP level, and procalcitonin level (p < 0.05). The total lesion score demonstrated the best performance when the data cut-off was 8.2%. Furthermore, the area under the curve, sensitivity, and specificity were 93.8% (confidence interval [CI]: 86.8–100%), 91.3% (CI: 69.6–100%), and 91.8% (CI: 23.0–98.4%), respectively. Conclusion CT quantitative parameters showed strong correlations with laboratory inflammatory markers, suggesting that CT quantitative analysis might be an effective and important method for assessing the severity of COVID-19, and may provide additional guidance for planning clinical treatment strategies.

Objective To explore the pancreatic kininogenase enteric-coated tablets in the treatment of diabetic retinopathy (DR) patients and its effect on the optic disc and macula retinal hemodynamics. Methods 86 cases (140 eyes) of DR patients were randomly divided into pancreatic kininogenase group and normal group with 43 cases in each group, two groups were treated with basic therapy, pancreatic kininogenase group were combined with pancreatic kininogenase treatment. The best corrected visual acuity and the clinical effect of optic disc and macula retinal hemodynamic changes were compared between two groups. Results After treatment, the best corrected visual acuity (BCVA) in pancreatic kininogenase group was greater than the normal group (P<0.05), the retinal neovascularization and fluorescein leakage area in pancreatic kininogenase group was less than the normal group (P<0.05). The disc vascular and macular retinal blood flow volume (VOL), blood flow velocity (FLW)values in pancreatic kininogenase group were larger than those of normal group, and the clinical curative effect of pancreatic kininogenase group was better than that of normal group (P<0.05). Conclusion Pancreatic kininogenase enteric-coated tablets in the treatment of DR patients can improve the optic disc and macular retinal hemodynamic parameters, improve visual acuity and reduce the retinal neovascularization and fluorescein leakage area, so as to improve the clinical treatment effect.

Objective To explore the pathogenesis of myelodysplastic syndrome (MDS)transformation, JAK2V617F mRNA expression levels were compared in peripheral blood of MDS-RA(refractory anemia,RA)and MDS-RAEB(RA with excess blasts,RAEB)patients.Methods JAK2V617F mRNA expression level was detected by fuorescent quantitative polymerase chain reaction(FQ-PCR),and this research reviewed 22 patients diagnosed with MDS,including outpatient and hospitalized patients.20 cases of normal control group were healthy ones.FQ-PCR method was applied to monitor JAK2V617F mRNA expression level in peripheral blood of MDS-RA and MDS-RAEB patients.Results The JAK2V617F gene was not expressed or expressed in healthy subjects,and the copy number was (3 851.96 ±470.46).The patients with MDS-RA(4 631.11 ±3 851.96)was significantly higher than that in healthy subjects(t =3.61,P <0.01);MDS-RAEB patients was (22 545.98 ±11 084.17),and was significantly higher than that in MDS-RA patients(t =4.87,P <0.01).Conclusion JAK2V617F signal transduction can play a role in the pathogenesis and transformation of MDS,and the detection of JAK2-V617F mRNA expression level is use-ful for monitoring progression,judging prognosis and gene regulation therapy of MDS.

Objective To explore the effect of irbesartan on cardiac endothelial-mesenchymal transition (EndMT) in diabetic rats.Methods The model of diabetic rat was induced by intraperitoneal injection with streptozotocin (STZ,35 mg/kg) in spontaneous hypertensive rats (SHR).Diabetic rats were divided into diabetic group and the Irbesartan treated group.The pathological changes were investigated by fluorescence microscope and electron microscope.The EndMT was studied in human aortic endothelial cells (HAEC) exposure to high glucose.The concentration of angiotensin Ⅱ in the supernatant was detected by radioimmunoassay.Immunofluorescence staining was performed to detect the co-localization of CD31 and FSP1.Results The significant myocardial fibrosis was presented in the diabetic group.Endothelial protrusions were prominent feature in myocardial microvascular of diabetic rat compared with the control group rats.Double staining of HAEC showed co-localization of CD31 and FSP1,which was decreased by the treatment of Irbesartan (P ＜ 0.05).When HAEC was exposed to high glucose,it showed some cells acquired spindle-shaped morphology and lost CD31 staining,and FSP1 and α-SMA protein expression levels were markedly upregulated,which attenuated by the treatment of Irbesartan.Conclusion Irbesartan might prevent diabetes from myocardial fibrosis via inhibition of EndMT in diabetic rats.

Objective To investigate the effects of low density lipoprotein receptor (LDLr) pathway on podocyte injury in diabetic nephropathy (DN) under inflammatory stress.Methods Male db/db mice and db/m mice were randomly divided into four groups (8 mice in each group):db/m group (control),casein injected db/m group (db/m + casein),db/db group (db/db),and casein injected db/db group (db/db + casein).An inflamed model of DN was established according to our previous study.24-hour urinary protein was measured every week.The plasma lipid profile was detected by clinical biochemistry assay.Podocyte changes were evaluated by electron microscope and immunofluorescent staining.Lipid accumulation in the kidney was evaluated by oil red O staining and intracellular cholesterol quantitative assay.The protein expression of Wilm's tumor-1 (WT-1),nephrin,α-smooth muscle actin (t-SMA),and molecules correlated with LDLr pathway were examined by immunohistochemical staining or Western blotting.The colocalized protein expression of LDLr with WT-1 was examined by immunofluorescent staining and laser confocal microscopy.Results There were no differences in plasma levels of LDL and HDL among four groups.Compared with db/db group,the db/db+ casein group showed markedly increased 24-hour urinary protein,more significant podocyte foot process effacement and podocyte damage,increased lipid droplet accumulation in kidneys,increased protein expressions of LDLr,SCAP and SREBP-2 in kidneys (all P ＜ 0.05).Interestingly,increased LDLr protein expression in kidneys of db/db mice was negatively correlated with decreased nephrin protein expression (r =-0.855,P ＜ 0.01) and positively correlated with increased α-SMA protein expression (r=0.768,P ＜ 0.01).Conclusions The disruption of LDLr pathway induced by inflammation contributes to podocyte injuries in diabetic nephropathy.

Objective To investigate the effect of human telomerase reverse transcriptase C27 polypeptide (hTERTC27) over-expression on orthotopic implanted tumor growth which induced by nasopharyngeal carcinoma cells C666-1 in vivo.Methods Stably transfected C666-1 cell lines were used to establish subcutaneously transplanted tumor mouse model.The tumor growth was observed and the tumor growth curve was made by measuring the volumes of tumors every day.HE staining was used to observe the change of histomorphology.The expressions of cleaved Caspase-3,Caspase-9,PARP,bax and bcl-2 were detected by Western blot analysis.Results The in vivo mouse model showed that over-expression of hTERTC27 significantly reduced tumor volume and tumor weight (P ＜ 0.05),and decreased the local muscle infiltration.Over-expression of hTERTC27 significantly up-regulated the expressions of cleaved Caspase-3,Caspase-9,PARP and bax (P ＜ 0.01),and down-regulated the expression of bcl-2 (P ＜ 0.01).Conclusion The results indicate that over-expression of hTERTC27 can obviously inhibit the growth of transplanted tumor in nude mice,which is possibly related to the regulation of bax and bcl-2 expression.

Objective To explore the clinical application and curative effect of iron removal treatment with week-therapy in chronic iron overload related to blood transfusion.Methods Twenty patients who were diagnosed as chronic iron overload were retrospectively studied.The amelioration of liver and pancreatic fanctions in patients with chronic iron overload treated with week-therapy of iron chelating agent through intravenous drip.Results The changes in the level of serum ferritin [2693.7±709.9) μg/L] were not significant after treatment at six months,hut the cilinical symptoms were relieved obviously.The levels of serum ferritin in patients after the treatment at twelve months were significantly different from the level before treated [(2083.7±714.4) μg/L vs (2771.5±725.2) μg/L,t =3.35,P ＜ 0.01],and the decreases were significant in sixteen patients.The liver functions in seven patients were improved and pancreatic functions were better in five patients.Conclusion The study suggests that iron removal treatment with week-therapy is effective for chronic iron overload,and improves the functions of damaged organs.

Objective To evaluate the effect and safety of the intermittent deferoxamine therapy on relieving iron overload caused by transfusion in senile. Method Twenty-three senile with iron overload caused by transfusion were administered in a total daily dose of 20 - 50 mg/ ( kg·d ) for 5 - 6 days per week every 4 -6 weeks,the maximal dose was 2000 mg/d, add in 0.9％ sodium chloride 500 ml, continuous intravenous drip for 6 h. Result Nine months after therapy, serum ferritin fell from (2771.5±735.3)μg/L to (2483.7 ±724.4) μg/L (P＜0.01), and urine ferritin elevated from (9.68 ±5.39)μg/L to (12.14±5.50) μ g/L (P＜0.01 ). Conclusion It shows that intermittent deferoxamine therapy can reduce the serum ferritin with no significant toxicity.