Objective To explore the method for early diagnosis of masked polycythemia vera (mPV) by comparing and analyzing the features of mPV and polycythemia vera (PV). Methods A total of 200 newly diagnosed male patients were collected based on diagnostic criteria, including 100 cases of PV and 100 cases of mPV. Erythropoietin (EPO), neutrophil alkaline phosphatase (NAP) score, bone marrow biopsy (BMB), and JAK2 V617F mutation were evaluated in all cases. After 6 months, hemoglobin (Hb) and JAK2 V617F mutation load were detected in patients without special treatment in two groups during follow-up. Results EPO, NAP score, BMB hematopoietic volume and the number of megakaryocyte had no statistical differences between PV group and mPV group [(3.4 ± 0.7) U/ml vs. (3.2±0.6) U/ml, (276±20) score vs. (278±21) score, (78±10) % vs. (76±9) %, (53±6) vs. (51±5), respectively], while JAK2 V617F mutation load in PV group was higher than that in mPV group[(89.2±9.4) % vs. (78.1±8.6) %, P<0.05]. In mPV patients without special treatment, Hb ≥185 g/L was found in 37 patients after 6 months, and the level of Hb and JAK2 V617F mutation load in these 37 patients reached (194±8) g/L and (90.7±9.1) %, respectively. Conclusions There is no significant difference in EPO, NAP score and BMB between PV and mPV, but the JAK2 V617F mutation load is different. In mPV patients without special treatment, Hb level can reach the typical PV diagnostic criteria after 6 months, meanwhile, JAK2 V617F mutation load is also increased.

Vascular endothelial growth factor,hypoxia inducible factor,interleukin,angiopoietin-like protein,integrin and epithelial mesenchymal transition can provide nutritional support and favorable environment for the growth,invasion and metastasis of cancer cells.Researches about mechanisms of the angiogenesis in esophageal squamous cell carcinoma may provide more ideas and potential targets for the anti-angiogenesis targeted therapy.

Objective To investigate the effects of exogenous glial cell derived neurotrophic factor (GDNF)on colon glial cells in slow transit constipation (STC ) rats,and to explore the optimal concentration of GDNF in order to provide evidence for intestinal neurotrophic therapy in the treatment of STC.Methods A total of 132 SD rats were divided into STC group and control group,66 rats in each group.STC rats were established by feeding with rhubarb.Six rats were randomly selected from either groups to verify whether STC model was successfully established.And the left 120 rats of two groups were randomly divided into six subgroups:STC group one to group six and control group one to group six,ten rats in each group,which were untreated,injected through tail vein with saline,and 0.001 ,0.010, 0.050,0.100 μg/L GDNF 2 mL respectively for one week.The expression of Sox-8 at protein level of either group were detected by Western blotting.Independent sample t test was performed for statistical analysis.Results After treated with 0.001 μg/L GDNF (STC group three),there was no significant
difference in expression level of Sox-8 between STC group three and STC group one (13.38 ±0.70 vs 13.39±0.45 ,t = 0.042,P = 0.969 ).After treated with 0.010 μg/L GDNF (STC group four),the difference in expression level of Sox-8 between STC group four and STC group three was significant (21 .11 ±2.56 vs 13.38±0.70,t=5 .040,P <0.01).After treated with 0.050 μg/mL GDNF (STC group five),the expression level of Sox-8 was higher than that in STC group four (31.86±1.57 vs 21.11±2.56,t=-6.198,P <0.01 ).The Sox-8 expression of untreated,saline treated,0.001 and 0.050 μg/L GDNF treated STC rats (STC group one,two,three and five)were lower than those of the corresponding control groups (t= 3.394,12.103,10.302,- 6.120,all P < 0.05 ).Conclusion Exogenous GDNF could increase Sox-8 expression in colon tissue of STC rats,an increase in the number of colon glial cells could repair enteric nervous system,and 0.050 μg/L was the optimal concentration.

Correspondinga uthor:Wan Chun, Email:mdchunwan@163.com
[Absrt act] Obj ective To summarize the clinical experience with isolated complete resection of he-patic caudate lobe in 14 patients. Mte hods The clinical data of 14patients with isolated complete resection of hepatic caudate lobe carried out in our hospital from December 2007 to March 2015 were retrospectively analyzed .During the operation , selective hepatic vascular occlusion slings , and supra-and infra-hepatic in-ferior vena cava slings were placed after full mobilization of the liver .Isolated complete resection of the he-patic caudate lobe was performed through the left and /or the right , the anterior liver-splitting or the retro-grade approaches .Results The mean operation time was (227 ±64) min.The mean amount of blood loss was (530 ±325) ml.The mean vascular occlusion time was (19.5 ±18.6) min.The mean diameter of ne-oplasm was (15 4.±9 .2) cm. All the operations were successfully carried out .There were no massive hem-orrhage, bile leakage or perioperative death .Hydrothorax occurred in 3 patients and ascites in 2.The mean stay in hospital was (21 ±9) days.All the patients were cured and discharged home .Conclusion Isolated complete resection of hepatic caudate lobe was feasible in clinical practice .

Objective To evaluate the safety of mesohepatectomy for centrally located large hepatic tumors.Methods The clinical data of 37 patients who underwent hepatectomy for centrally located large liver tumors in our hospital from October 2010 to August 2015 were retrospectively analyzed.During the operation,the left and right hemilivers were mobilized.Slings for the improved liver hanging maneuver and for selective hepatic vascular occlusion were placed.These slings were used when necessary in order to minimize occlusion and ischemia time to the residual liver,and to maximize the volume of functional liver remnant.Mesohepatectomy was carried out using a microwave hemostatic separator.The feasibility,its effects on preventing hemorrhage,the degree of liver damage and the postoperative complications were evaluated.Results The mean diameter of the neoplasms was (12.6 ± 7.2) cm.The vascular inflow occlusion time of the left and right hemilivers were (12.2 ±3.5) min and (18.5 ±7.1) min,respectively.The blood loss was (487 ± 352) ml.The amount of red blood cell transfusion was (2.7 ± 1.9) U.The operation time was (215 ± 72) min.TBIL,ALT and AST reached their peak levels on the 1 st day after operation and they were higher than before surgery (P ＜ 0.01).The levels were then significantly decreased on the 3rd day after operation.However,the TBIL (P ＜ 0.05),ALT (P ＜ 0.01) and AST (P ＜ 0.05) were higher than the preoperative level,and only returned to normal or were close to the preoperative value on the 7th day after operation (P ＞ 0.05).The initial increase in ALT was quick and then it slowed down,while the initial increase in AST was slow but it decreased quickly.Conclusions Mesohepatectomy for centrally located large hepatic tumors could safely be carried out using the liver hanging maneuver combined with selective hepatic vascular occlusion and a microwave hemostatic separator.This method has the advantages of causing less bleeding,liver damage and rapid recovery.

Objective To explore the effects of calcitonin gene-related peptide (CGRP) on proliferation of vascular endothelial cells (VECs) and osteogenic differentiation of bone mesenchymal stem cells (BMSCs) of osteoporotic rats in vitro,and cell/ molecular mechanism of promoting osteoporotic fracture healing.Methods 10 3-month-old SD rats were employed to make osteoporosis model by taking bilateral ovariectomy.Micro-CT analysis was used to determine the success of modeling at 6 months of postoperation.BMSCs and VECs were isolated from osteoporotic rats by blood adherence mcthod.10-7-10-10 mol/L CGRP were added to the culture medium.CCK-8 method was used to examine the cell proliferation.Effect of CGRP on osteogenic differentiation of BMSCs:Alkaline phosphatase (ALP) activity,and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of ALP,Collagen Ⅰ,BMP-2,Osteonectin and RunX2 gene.Effect of CGRP on vascularization of VECs:Vascular cell endothelial growth factor (VEGF) protein secretion and gene expression level were tested by ELISA and RT-PCR.Resuits BMSCs and VECs of osteoporotic rats were isolated successfully.The optical density (OD) values of CCK-8 method increased in a dose-dependent manner,for BMSCs at 7 d,the control group of OD value was 0.58±0.02,the CGRP group 10-7-10-10 mo]/L was to 0.80±0.03,0.79±0.03,0.74±0.03,0.69±0.03,difference was significant;for VECs,the control group of OD value was 2.20±0.01,the CGRP group 10-7-10-10 mol/L was to 4.12±0.04,3.90±0.02,3.56±0.04,3.12±0.04,difference was significant.Effect of CGRP on osteogenic differentiation of BMSCs:Alkaline phosphatase (ALP) activity,and reverse transcription polymerase chain reaction (RT-PCR) were used to detect the expression of ALP,Collagen Ⅰ,BMP-2,Osteonectin and RunX2 gene.Effect of CGRP on vascularization of VECs:Vascular cell endothelial growth factor (VEGF) protein secretion and gene expression level were tested by ELISA and RT-PCR,difference between the control group and CGRP group were all significant.Conclusion 10-7 mol/L,10-10 mol/L of CGRP can directly promote the proliferation of the BMSCs and VECs of osteoporotic rats,promote the osteogenic differentiation of BMSCs,improve the expression of VEGF gene and protein secretion,promote angiogenesis.

Objective To investigate the efficiency of intraoperative neurophysiological monitoring in cervical spinal tumor surgery.Methods Retrospective case-control study comparing 23 cervical spinal tumor patients received surgery under neurophysiological monitoring (case) and 23 cases without neurophysiological monitoring (control).Results All of 46 cases,there were no significant differences of age,gender and preoperative neurological function.Duration of operation and estimated blood loss in the IONM cohort was less than the control group with significant difference.The preoperative JOA score of IONM group and control group were 12.0±2.4 and 12.7±2.1,with no significant difference.The postoperative JOA score of each group were 15.0±1.5 and 15.2±1.7,with no significant difference.The rate of cervical cord neurological improvement of IONM group was higher than control group,but there was no significant difference.In the IONM cohort,abnormal signal appeared in 8 cases,with 1 case developed new postoperative deficit,and the sensitivity and specificity of SEP were 100％ and 77.3％,the sensitivity and specificity of MEP were 100％ and 86.4％.Compared with preoperative SEP,4 cases showed major improvement (baseline increase ＞ 40％),2 cases showed moderate improvement (baseline increase:15％-40％).The coincidence rate of the SEP improvement and the AISA grade was 66.7％.Conclusion IONM could reduce the duration of operation and intraoperative blood loss.Changes of IONM were correlated with postoperative neurological function improvement.

Objective To determine the reference values of 11 amino acids measured by non-derivatized tandem mass spectrometry ( MS/MS).Methods 22 430 healthy newborns and 14 children with metabolic dysfunction in Nanjing were enrolled in this study.The levels of the 11 amino acids, including Ala, Arg, Cit, Gly, Leu +Ile, Met, Orn, Phe, Pro, Tyr and Val, were measured by non-derivatized tandem mass spectrometry using dry blood spots.After pre-setting up different cutoff values according to the amino acid levels of the newborns, reference value ranges of the 11 amino acids were determined by using the method of percentile combined with ROC curves.Results According to the results of frequency distribution histogram, the levels of the 11 amino acids of newborns belong to approximate normal distribution.By analysis of ROC curves, the cut-off values for amino acids were 0.2%-99.8%.Conclusion It established reference values of 11 amino acids in newborn and provided basis for the other screening center to make the cut-off value.

Objective:To explore the application of high-throughput sequencing (HTS) technology in pathogens detection for spinal infection.Methods:From January 2019 to May 2020, a total of 41 patients including 31 males and 10 females with an average age of 59.7±11.9 years (29-75 years) were suspected of spinal infections. There were 37 patients with local pain, 15 with fever (≥38 ℃) and 18 with neurological dysfunction. The infected sites were as follows, 4 cases of cervical spine, 8 cases of thoracic spine and 29 cases of lumbar spine. There were 36 patients met the surgical indications and underwent open debridement, bone grafting, fusion and internal fixation, while the other 5 patients underwent conservative treatment (three received drug therapy and two were transferred to the internal department for chemotherapy). Lesions obtained from open surgery patients were underwent pathology and HTS examination. In 5 cases with conservative treatment, two of them underwent CT guided percutaneous puncture for samples, while one case underwent ultrasound guided percutaneous puncture for pus, one case for venous blood, and one case received lumbar puncture for cerebrospinal fluid. The samples were sent for pathological and HTS examination, while liquid specimens were sent for bacterial culture and HTS. The sensitivity and specificity of HTS results were determined according to pathological examination which was regarded as the "gold standard". Based on HTS results combined with the clinical manifestations, imaging examination and pathological results of the patients, targeted antibiotics or anti-tuberculosis drugs were selected for postoperative drug therapy. Patients with bacterial infection received anti-infection treatment for 3 months after operation. For tuberculosis patients, "tetrad" (isoniazid+rifampicin+pyrazinamide+ethambutanol) anti-tuberculosis treatments were underwent for one year. Inflammation indicators from the blood samples were observed before and after treatment, including white blood cell count (WBC), C-reactive protein (CRP) and erythrocyte sedimentation rate (ESR). These indicators were used to monitor disease progression and the curative effects. All patients were followed up for at least 3 months after surgery.Results:A total of 41 patients with suspected spinal infection were included in this study. The HTS pathogen detection results were obtained within 48 h. For the initial 5 patients, first-generation sequencing verification was conducted with coincidence rate 100%. Further, no further verification was conducted in the rest patients. Among the 41 cases, a total of 26 cases had positive results with a positive rate of 63.4%(26/41). Among them, thirteen cases were with mycobacterium tuberculosis (31.7%) and 6 cases with staphylococcus (14.6%). Fungi and Brucellosis were diagnosed in 2 cases respectively, accounting for 4.9% respectively. The test were negative in 15 patients (36.6%), including 2 patients with tumor or tumor-like lesions (1 hematologic tumor and 1 eosinophilic granuloma). A total of 38 patients underwent pathological examination, which confirmed 7 cases of suppurative infection, 12 cases of tuberculosis, 2 cases of tumor or tumor-like lesions and the remaining 17 cases of inflammatory lesions. The sensitivity and specificity of HTS were 80%(16/20) and 55.6% (10/18) with positive predictive value (PPV) 66.7% (16/24) and negative predictive value (NPV) 71.4% (10/14). All patients were followed up for 3 months. The inflammation indicators of blood at 3 months were all lower than that at admission. WBC decreased from (7.50±3.26)×10 9/L at admission to (6.22±2.53)×10 9/L at 3 months after treatment without statistically significant difference ( t=1.082, P=0.290). The CRP decreased from (32.2±34.1) mg/L to (4.5±10.5) mg/L, and ESR from (44.2±26.5) mm/1 h to (18.6±12.1) mm/1 h with statistically significant difference ( t=8.963, P<0.001; t=5.421, P<0.001). Conclusion:High-throughput sequencing technology can be used in detection of spinal infection pathogens, due to its relatively high positive rate, satisfied sensitivity and good diagnostic value.

Objective: To investigate the long-term outcomes of posterior long segment instrumentation and fusion in adult degenerative scoliosis (ADS), and to explore the correlative factors of sagittal imbalance after long segment instrumentation and fusion, and to summarize the impacts of different lowest instrumented vertebra (LIV) on sagittal complication rates. Methods: ADS patients who underwent long segment instrumentation and fusion between January 2008 and January 2014 were retrospectively reviewed and divided into two groups depended on LIV (L ₅ group and S₁ group). The follow-up time was at least five years. Radiographic parameters, Visual Analogue Score (VAS), Oswestry Disability Indexscore(ODI), the 12-item Short Form Health Survey-Physical Component Summary (SF-12 PCS)and sagittal complications were analyzed. And the correlation between sagittal imbalance and sagittal complications were calculated. Patients were divided into two groups(sagittal imbalance group and sagittal balance group) to explore the correlative factors of sagittal imbalance. Results: All of 56 patients were included and evaluated in this study, and 35 cases stopped at L ₅, 21 cases at S₁. The mean follow-up time was 6.6±1.6 years. No significant difference were found in age, gender, follow-up time and surgery method (P>0.05) between two groups. No significant differences were found in Cobb angle(11.8°±7.3° vs 12.6°±6.4°), coronal balance distance(CBD) (0.7±0.6 cm vs 1.0±0.5 cm), sagittal vertebral axis(SVA)(3.0±3.1 cm vs 2.6±5.6 cm), thoracic kyphosis(TK) (21.4°±9.6° vs 22.5°±6.9°), lumbar lordosis(LL) (-32.7°±7.3° vs-34.8°±9.1°), pelvic tilt (PT) (23.4°±9.8° vs 23.3°±8.1°), VAS score(2.0±1.2 vs. 2.2±0.9), ODI score(18.5%±6.4% vs 19.3%±5.2%) and SF-12 PCSscore(45.5±5.2 vs 42.4±7.0) between two groups at final follow-up time(P>0.05). Radiographic parameters including Cobb angle, CBD, SVA and health related quality of life scores including VAS score, ODI score and SF-12 PCS scores were improved significantly at two weeks postoperatively, two-year follow-up time andfinal follow-up time in two groupscompared with those preoperatively (P<0.05). And no significant difference were found in radiographic parameters and health related quality of life scores between those at two years follow-up time and those at final follow-up time; Sagittal complication rates at final follow-up were higher than these at two years follow-up (82.1% vs 33.9%, P=0.000). Sagittal imbalance was related to internal fixation failure in L ₅ group (r=0.691, P=0.011), internal fixation failure in S₁ group (r=0.691, P=0.001) and pseudoarticulation formation (r=0.499, P=0.021). 10 patients in the sagittal imbalance group were with preoperative sagittal imbalance, 9 with preoperative coronal imbalance, 6 with internal fixation failure, 5 with proximal junctional kyphosis (PJK), 4 with adjacent segment degeneration during the follow-up periods, the incidence rates of which were higher than those in the sagittal balance group. LL in the sagittal imbalance group was lower than that in the sagittal balance group at two years follow-up time and final follow-up time. Compared with the sagittal imbalance group, the sagittal balance group achieved a higher SF-12 PCS (45.4±5.1 vs 41.6±7.4, P<0.05) and lower ODI scores (17.8%±6.1% vs 21.5±4.6%, P<0.05), and no significant difference was found in VAS scores between two groups (1.9±1.2 vs 2.4±0.8, P>0.05). The incidence of adjacent segment degeneration in L ₅ group was higher than that in S₁ group (P<0.05) at final follow-up time, and the incidence of pseudojoint in S₁ group was higher than that in L ₅ group (P=0.002) at final follow-up time. Conclusion Posterior long segment instrumentation and fusion for adult degenerative spinal deformity can obtain satisfying long-term clinical outcomes, and postoperative sagittal imbalance was related to preoperative sagittal/coronal imbalance, postoperativeinternal fixation failure, PJK, pseudoarticulation formation and adjacent segment degeneration. Distal fusion at L ₅ or S₁ may lead to high risk of adjacent segment degeneration or pseudoarticulation formation respectively.