Objective:To explore the values of peripheral blood neutrophil-to-lymphocyte ratio (NLR), monocyte-to-lymphocyte ratio (MLR) and platelet-to-lymphocyte ratio (PLR) in the differential diagnosis of cytopenic diseases.Methods:A retrospective case series study was conducted. The clinical data of 105 newly diagnosed patients with aplastic anemia (AA), myelodysplastic syndrome (MDS) or primary immune thrombocytopenia (ITP) who were admitted to Nanjing Drum Tower Hospital, the Affiliated Hospital of Nanjing University Medical School from August 2017 to November 2020 were retrospectively analyzed. There were 42 patients with MDS (13 cases of hypoplastic MDS, 23 cases of non-hypoplastic MDS, 6 cases could not be classified), 42 patients with ITP, and 21 patients with AA. The peripheral blood lymphocyte count, neutrophil count, monocyte count, and platelet count of each untreated patient at the time of initial diagnosis were recorded, the NLR, MLR and PLR were calculated, and the differences of NLR, MLR and PLR among different diseases were compared; the differential diagnostic values of each index for AA, MDS and ITP were evaluated by using the receiver operating characteristic (ROC) curve.Results:The NLR [ M (IQR)] in ITP, AA and MDS groups was 3.08 (2.42), 0.57 (0.66) and 0.83 (1.27) ( χ2 = 56.84, P<0.001), the MLR was 0.26 (0.15), 0.13 (0.14) and 0.20 (0.33) ( χ2 = 18.71, P<0.001), and the PLR was 5.12 (9.97), 8.67 (14.21) and 49.32 (78.66) ( χ2 = 47.07, P<0.001). The best cut-off value of NLR for distinguishing ITP from MDS was 1.757, and the area under the curve (AUC) was 0.833 (95% CI: 0.811-0.955), while the sensitivity and specificity were 78.6% and 83.3%, respectively. The best cut-off value of NLR for distinguishing ITP from AA was 1.350, and the AUC was 0.993 (95% CI: 0.981-1.000), while the sensitivity and specificity were both 95.2%. The best cut-off value of PLR for distinguishing AA from MDS was 23.542, and the AUC was 0.841 (95% CI: 0.739-0.944), while the sensitivity and specificity were 85.7% and 73.8%, respectively. The best cut-off value of NLR for distinguishing AA from MDS was 0.764, and the AUC was 0.687 (95% CI: 0.556-0.891), while the sensitivity and specificity were 71.4% and 64.3%, respectively. The best cut-off value of MLR for distinguishing AA from MDS was 0.148, and the AUC was 0.736 (95% CI: 0.614-0.859), while the sensitivity and specificity were both 71.4%. Conclusions:The peripheral blood NLR, MLR and PLR have certain reference values for differential diagnosis of AA, MDS and ITP.

Objective:To investigate the molecular pathogenesis of two coagulation factor Ⅺ (FⅪ) deficiency patients.Methods:Coagulant assays: activated partial thromboplastin time(APTT), normal pooled-plasma corrected APTT test, PT, PT-INR and one-stage assay of coagulation factors activities were validated to diagnose coagulation factor Ⅺ deficiency. The patients’ DNA samples were extracted and all exons and flanking sequences of F11 gene were amplified using PCR. After purified, the products of PCR were sequenced directly, the mutations were detected by comparing with wild sequences and analyzed using some bio-informatics softwares. Results:The two patients were diagnosed with coagulation factor Ⅺ deficiency due to prolonged APTT, corrected APTT and low activities of coagulation factor FⅪ. The results of APTT, FⅪ∶C were 88.1s, 1.1% and 107.1s, 3.8% , and the prolonged APTT could be corrected to normal range 32.9s and 31.5s, respectively. Through genetic analysis, we discovered compound heterozygous mutations g. 1305-1G>A and g. 1325delT in patient 1 and the sequencing results of TA plasmid clones showed that the two mutations were located on different strands of chromosomes. Compound heterozygous mutations g. 1124A>G and g. 1550C>G were detected in patient 2 resulting in Lys357Arg and Cys482Trp. Software analysis indicated the mutations probably brought amino acid sequence changed, protein features affected and splice site changed.Conclusion:Compound heterozygous mutations g. 1305-1G>A, g. 1325delT and g. 1124A>G, g. 1550C>G had been identified in two coagulation factor Ⅺ deficiency patients which might be responsible for their prolonged APTT and low FⅪ∶C. To the best of our knowledge, g. 1325delT and g. 1550C>G have been reported, while g. 1124A>G and g. 1305-1G>A are reported for the first time in the literature.

Objective:To investigate the expression of microRNA-324-5p (miR-324-5p) and transcription factor forkhead box C1 (FOXC1) in glioma and their relationship with the prognosis of patients.Methods:From March 2012 to March 2015, a total of 72 cases of glioma tissues were collected from glioma patients who were admitted to Chongqing Hygeia Tumor Hospital and the People's Hospital of Nanchuan in Chongqing, and 28 cases of normal human brain tissues resected in craniocerebral surgery were also collected. The expressions of miR-324-5p and FOXC1 mRNA were detected by real-time fluorescent quantitative polymerase chain reaction (qRT-PCR), and the expression of FOXC1 protein was detected by immunohistochemistry. Pearson method was used to analyze the correlation between the expressions of miR-324-5p and FOXC1 in glioma tissues; Kaplan-Meier method was used to analyze the survival of patients with glioma; Cox regression analysis was used to analyze the risk factors affecting the prognosis of patients with glioma.Results:FOXC1 protein was mainly located in the cytoplasm of glioma, and its positive expression rate in glioma tissues was 81.94% (59/72), which was significantly higher than that in normal brain tissues [17.86% (5/28)], and the difference was statistically significant ( χ2 = 35.938, P<0.01). Compared with normal brain tissues, the expression of miR-324-5p was down-regulated in glioma tissues (0.62±0.19 vs. 0.98±0.02, t = 9.974, P < 0.05), and the expression of FOXC1 mRNA was up-regulated (1.41±0.29 vs. 0.99±0.02, t = 7.633, P < 0.05). The expressions of miR-324-5p and FOXC1 protein were correlated with the number of primary lesions, differentiation degree, TNM stage and lymph node metastasis of glioma (all P<0.05). Pearson analysis showed that the expressions of miR-324-5p and FOXC1 mRNA were negatively correlated ( r = -0.550, P<0.01). The 5-year overall survival rate of patients in miR-324-5p high-expression group was significantly higher than that of patients in miR-324-5p low-expression group (45.71% vs. 24.33%, χ2 = 6.531, P = 0.011), and the 5-year overall survival rate of patients in FOXC1 protein high-expression group was significantly lower than that of patients in FOXC1 protein low-expression group (30.41% vs. 42.34%, χ2 = 3.631, P = 0.047). Multivariate Cox regression analysis showed that low differentiation, TNM stage Ⅲ-Ⅳ, lymph node metastasis, low expression of miR-324-5p and high expression of FOXC1 protein were independent risk factors for prognosis of glioma patients (all P < 0.05). Conclusions:The expression of miR-324-5p is low and the expression of FOXC1 is high in glioma. They may be involved in the regulation of tumor differentiation and metastasis, and related to the poor prognosis of patients. They may be potential therapeutic targets for glioma.

Objective To retrospectively analyze the platelet count and related factors in bleeding patients with hematonosis,and to calculate the risk of bleeding when the platelet count is at each exposure level.Methods Retrospective analysis of patients from Department of Hematology Inpatients in Nanjing Drum Tower Hospital,Nanjing First Hospital and Nanjing Jiangning Hospital from July 2013 to June 2017 was collected.And the risk of bleeding for different hematonosis was calculated.Results The tolerance of the 5 categories of hematonosis to low platelet counts is compared:AA and ITP can tolerate lower levels of platelet count;MDS and AML(except M3) are more prone to bleeding;ALL is the most susceptible to bleeding.Conclusion When platelet resources are scarce,priority should be given to ALL,MDS and AML patients,in order to ensure the safety of critically ill patients.For patients with AA and ITP,the platelet infusion threshold may be reduced appropriately,in oder to reduce the incidence of platelet transfusion refractoriness.

Objective To investigate the molecular etiology of protein C (PC) deficiency.Methods Routine diagnosis and genetic analysis were performed on four probands with PC deficiency.Results ①Case 1,female,40 years old,diagnosed of deep vein thrombosis in left lower limb.PC activity (PC∶ C) was 48％,PS activity (PS∶ C) was 26.3％,AT activity (AT∶ C) was 75.6％.Genetic analysis discovered heterozygous mutation C5156T on promoter of PC gene,together with heterozygous mutation A6578T on Exon2 of PC gene.After anticoagulant,thrombolysis and filter implantation therapies,the patient went home with improvement.②Case 2,female,32 years old,diagnosed of deep vein thrombosis in both lower limb,ischemia in both lower and upper limb,and skin infection in both lower limb.PC∶ C 27％,PS∶ C 22.9％,AT∶ C 86.7％.Genetic analysis identified heterozygous mutation C5156T,together with heterozygous mutation A5045T on promoter of PC gene.After anticoagulant and anti-infection therapy,the patient died of respiratory failure,septic shock and DIC.③Case 3,female,28 years old,diagnosed of vein thrombosis in right iliac and femoral vein.PC∶ C 58％,PS∶ C 57.3％,AT∶ C 80.8％.Genetic analysis disclosed heterozygous mutation C4867T on promoter of PC gene,AGA 12702-12704del or 12705-12707del on Exon7,the latter one lead to Arg192 or 193del.Heterozygous mutation G15240A on Exon9 was also found.After anticoagulant,thrombolysis and filter implantation therapies,the patient went home with improvement.④Case 4,male,30 years old,diagnosed of vein thrombosis in right iliac and femoral vein.PC∶C 50％,PS∶C 75.0％,AT∶C 89.1％.Genetic analysis found homozygous mutation C4867T and G4880A on promoter of PC gene,heterozygous mutation A5045T on promoter and heterozygous mutation T6589C on Exon2.After anticoagulant,thrombolysis and filter implantation therapies,the patient went home with improvement.⑤Polymorphism analysis revealed that heterozygous mutation C4867T,homozygous mutation G4880A,and heterozygous mutation C5156T were polymorphism sites of PC gene.Conclusions Polymorphism sites (G4880A,C4867T,C5156T),missense mutation A5045T,A6578T,G15240A,and deletion mutation AGA12702-12704del,12705-12707del may be related to deficiency of PC.Missense mutation A5045T,A6578T,G15240A,and deletion mutation AGA12702-12704,12705-12707del were first reported worldwide.

Objective To discuss the medium-term follow-up of clinically insignificant residual fragments (CIRF) after minimally invasive percutaneous nephrolithotomy lithotripsy (MPCNL).Methods The clinical data of 72 patients with CIRF medium-term follow-up were analyzed retrospectively.Results Seventy-two patients with CIRF.The anatomical distribution of CIRF was 10 at upper pole,15 at middle,35 at lower,10 at renal ureteropelvie junction and 2 at upper and lower pole.Stone analysis showed that 41 cases of calcium oxalate calculi,16 of calcium oxalate calculi mixed with carbonate calculi,3 calcium oxalate calculi mixed with uric acid,4 calcium oxalate calculi mixed with struvite stone,3 struvite stone,2 uric acid stone and 3 carbonate apatite mixed with struvite stone.Fifteen cases had clinical symptoms,including 2 renal colic pain,8 hematuria,5 lower urinary tract symptoms,4 cases CIRF located in upper pole,1 case in middle pole,4 cases in lower pole,6 cases in ureteropelvic junction,the incidence of clinical symptoms in ureteropelvic junction was significantly higher than that in other locations (6/10 vs.4/12,1/15,4/37,P ＜0.05).Eight cases required surgical procedure,5 cases underwent extracorporeal shock wave lithotripsy,3 cases with ureteral CIRF were performed with ureteroscopic lithotripsy.CIRF were clear after surgery,7 patients with ureteral CIRF had renal colic pains.The stones were excluded after spasmolytic analgesic treatments.Conclusions CIRF can be located variously in the kidney and ureter.Most CIRF are calcium oxalate calculi and locate in the lower pole.Patients with the history of previous open surgery or extracorporeal shock wave lithotripsy are more likely to get CIRF.Medium-term follow-up of CIRF reveals that CIRF located in the renal ureteropelvis junction are more likely to have clinical symptoms.

Objective To evaluate the clinical effects and safety of percutaneous nephrolithotomy (PCNL) by middle renal calice used as the main target for the treatment of staghorn stones with the combination of pneumatic and ultrasonic lithotrite.Methods Clinical data of 73 patients underwent PCNL by middle renal calices as main access with 57 incomplete staghorn stones and 35 complete staghorn stones.To observe the situation calculus removal rate and complications.Results Seventy cases (88 sides) underwent one session PCNL by single access tract (middle caliees),3 cases (4 sides) underwent one session PCNL by double access tracts (2 cases by middle and low calices,1 case by up and middle caliees).After the first period of lithoclasty,17 patients (25 sides) residual stones and the stone removal rate 72.8％ (67/92),among these patients,1 case (1 side) had fragments of lateral renal calyeeal stones with no further treatment.Other 16 cases (24 sides)underwent second session PCNL,all were treated by single access tract (middle calices) and 2 cases (2 sides)had extracorporeal shock wave lithotripsy before the second PCNL.After the second period of lithoclasty,76 sides composed of 27 complete staghorn stones and 49 incomplete staghorn stones had no residual fragments with the stone removal rate 82.6％ (76/92).The operative time lasted 120-320 min.Hemoglobin dropped 1-4 g/L,11 cases in the operation procedure and 3 cases after operation needed blood transfusion respectively.One case of renal pelvic infection after operation and 1 case had split renal dysfunction with peri-parenchyma infection.The hospitalization time was 9-18 days.Conclusion It is effective and safe to perform PCNL for staghorn stone by middle calices as a main access.Combining pneumatic and ultrasonic lithotrite will be very useful with high stone clearance,short procedure time and less complications.

Objective To observe the effect of bone marrow-derived mesenchymal stem cells (bMSCs) on graft healing within a bone tunnel after anterior cruciate ligament (ACL) reconstruction in rabbits.Methods The study involved 24 New Zealand white rabbits undergone ACL reconstruction with an autologous ipsilateral gastrocnemius tendon graft.Both hindlimbs were included.In one hindlimb,graft coated with fibrin glue compound by bMSCs was employed (bMSCs group).Whereas in the contralateral hindlimb,graft coated with fibrin glue without cells was employed (control group).At postoperative 2,4,6 and 8 weeks,specimens were harvested to have a biomechanical test of tensile strength and stiffness of tendon-bone interface.Results Tensile strength and stiffness of tendon-bone interface in both experiment and control groups presented a rising trend with the prolong of repair time.In contrast,significantly higher tensile strength and stiffness of tendon-bone interface were observed in experiment group since the 6 weeks (P ＜ 0.05).Conclusion bMSCs transplantation significantly enhances the early tensile strength and stiffness at tendon-bone interface after ACL reconstruction in rabbits and improves the graft healing within a bone tunnel.

Objective To compare the outcomes of antegrade and retrograde approach ureteroscopy for impacted upper ureteric calculi and assess the safety and efficiency of the two types of minimally invasive technique. Methods A total of 106 patients with impacted upper ureteric calculi were treated with ureteroscopy. The procedure was performed via antegrade percutaneous nephrostomy tract in 50 patients (antegrade group) and via retrograde transurethral access in 56 patients (retrograde group). Results The success rate of retrograde group was 92.9% (52/56). Operating time was (45 ± 5 ) min, hospital stay was (6 ± 1) days. The stone free rate was 80.4%(45/56) at 1 month follow-up,7 patients with residual calculi required ESWL combination. Complication rate was 5.4% (3/56). The success rate of antegrade group was 100.0% (50/50). Operating time was (55 ± 8 ) min, hospital stay was (8 ± 2) days. The stone free rate was 100.0% (50/50) and no complication was noted. The stone free rate and the complication rate indicated significant difference between the two groups (P ＜ 0.05). Conclusions Antegrade and retrograde access ureteroscopy for impacted upper ureteric calculi are safe and effective. Success rate and stone free rate of antegrade approach are higher than those of retrograde approach.

Juvenile hemochromatosis is an autosomal recessive disease characterized by progressive tissue iron overload which leads to irreversible organ damage and even death.This disease is mainly caused by mutations in two genes:hemojuvelin gene and hepcidin gene.Different mutations have different phenotype.The two genes may act as modifying genes in HFE hemochromatosis.Hepcidin secreted by liver plays a central role in the regulation of iron homeostasis.HJV can act as a bone morphogenetic protein(BMP)co-receptor which is required for HJV to regulate hepcidin expression and iron homeostasis.Recent researches suggest that the bone morphogenetic protein(BMP)signaling pathway mediated by HJV is a significant mechanism for HJV to regulate hepcidin expression and iron homeostasis.HJV mutant impaires BMP signaling which results in hepcidin expression decrease and abnormal iron metabolism.