Ankylosing spondylitis (AS) combined with lower cervical fracture is often categorized into unstable fracture, with a high incidence of neurological injury and a high rate of disability and morbidity. As factors such as shoulder occlusion may affect the accuracy of X-ray imaging diagnosis, it is often easily misdiagnosed at the primary diagnosis. Non-operative treatment has complications such as bone nonunion and the possibility of secondary neurological damage, while the timing, access and choice of surgical treatment are still controversial. Currently, there are no clinical practice guidelines for the treatment of AS combined with lower cervical fracture with or without dislocation. To this end, the Spinal Trauma Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate Clinical guidelines for the treatment of ankylosing spondylitis combined with lower cervical fracture in adults ( version 2024) in accordance with the principles of evidence-based medicine, scientificity and practicality, in which 11 recommendations were put forward in terms of the diagnosis, imaging evaluation, typing and treatment, etc, to provide guidance for the diagnosis and treatment of AS combined with lower cervical fracture.

The heart valve prosthesis must have excellent hydrodynamic performance which is usually tested in vitro, not in vivo. This paper comprehensively introduced the principles and methods of hydrodynamic performance in vitro testing, helping clinicians to understand valve performance parameters, evaluate valve applicability, and reduce clinical risk of the valve prosthesis. In vitro testing not only serves as the "gold standard" for valve prosthesis assessment, but also provides detailed data for design and optimization of the prosthesis. ISO 5840 defines the items and methods for valve in vitro testing, which consists of three parts: (1) pulsatile flow testing, which reproduces the pulsating flow of the valve prosthesis after implantation in the human body; (2) steady flow testing, which assesses valve forward flow resistance; (3) durability testing, which evaluates the durability of the valve prosthesis and determines the expected failure   mode. In addition, the paper presented the differences between atrioventricular and aortic valve testing, the method of mitral valve testing, the differences between transcatheter and surgical valve testing, and the method of valve flow visualization.

Objective:To investigate the relationship between the serum alkaline phosphatase (ALP), prostate specific antigen (PSA) and bone metastasis in initially diagnosed prostate cancer (PCa) patients in different ISUP(International Society of Urological Pathology)groups.Methods:The 368 initial diagnosed prostate cancer patients recruited from January 2013 to December 2018 were retrospectively analyzed, including 247 cases in the Third Affiliated Hospital of Sun Yat-sen University, 111 cases in the Yuebei People's Hospital Affiliated to Medical College of Shantou University and 10 cases in Shenzhen Hospital of Southern Medical University. According to whether there was bone metastasis at the initial diagnosis, it was divided into 230 cases in the bone metastasis group and 138 cases in the non bone metastasis group. There was no significant difference between the two groups in age [(71.9±9.4) years and (71.2±8.7) years], body mass index (BMI) [(23.1±3.7) kg/m 2 and (23.7±2.6) kg/m 2]. There were significant differences in PSA [(307.3±847.0) ng/ml and (84.5±257.3) ng/ml] and ALP [(174.5±270.8) U/L and (71.0±23.2) U/L] between the two groups. In different PSA subgroups, there were 45 cases in PSA <10 ng/ml, 35 cases in PSA 10-20 ng/ml and 288 cases in PSA >20 ng/ml. The differences of ALP and PSA between bone metastasis group and non-bone metastasis group based on different ISUP stratification were analyzed, ROC curves were used to predict their risks of bone metastasis. Results:There were 3(1.3%), 22(9.6%), 34(14.8%), 85(37.0%) and 86 (37.4%) prostate cancer patients with bone metastasis from ISUP group 1 to 5, and 14(10.1%), 19(13.8%), 29(21.0%), 32(23.2%) and 44(31.9%) without bone metastasis, respectively. There was significant difference in the serum ALP levels between the bone metastasis group and the boneless metastasis group in the ISUP group 4(157.6±207.7 vs. 66.5±17.0) and 5(189.4±257.5 vs. 69.2±18.4)( P<0.001) and PSA levels had difference in the ISUP group 3(240.3±313.0 vs. 42.4±42.1), 4(152.3±184.5 vs. 44.7±33.3) and 5(435.2±1006.3 vs. 60.8±84.8)( P<0.001). There was statistically significant between the bone metastasis group and the without(336.1±882.2 vs. 139.3±328.1) when PSA>20 ng/ml( P=0.006). ROC curve analysis: the cut-off values of ALP were 115.5, 109.0, 75.5 and 86.0 U/L from ISUP group 2 to 5 respectively, the sensitivity was 23.8%, 56.5%, 66.4% and 50.6% respectively, the specificity was 99.7%, 93.4%, 78.3% and 89.2% respectively, and the accuracy were 59.4%, 73.1%, 69.7% and 63.3%, respectively. The cut-off values of PSA were 39.5, 93.1, 54.2 and 28.9 ng/ml from ISUP group 2 to 5 respectively, the sensitivity was 64.4%, 68.4%, 87.4% and 88.3% respectively, and the specificity was 79.5%, 90.6%, 63.7% and 61.6% respectively, and the accuracy were 71.6%, 78.1%, 80.1% and 79.2%, respectively. Conclusion:ALP increased significantly in ISUP group ≥4 and PSA in ISUP group ≥3, which related to bone metastasis in patients with initial diagnosed prostate cancer.

Objective: To investigate the clinicopathological features and molecular phenotypes of gastric cancer with enteroblastic dif-ferentiation (GCED). Methods: A retrospective analysis of 337 patients with gastric adenocarcinoma diagnosed by the pathology de-partment of the First Affiliated Hospital of Zhejiang University in March 2013-2017 was conducted. Of them, 8 patients were diag-nosed with gastric carcinoma with intestinal blastocyte differentiation. All the patients were elderly, including 6 men and 2 women. The onset age was 68-83 years (mean 76.6 years). Two cases had serum AFP≥200 μg/L before treatment. According to the histopatho-logical morphology, the immunophenotype was analyzed by immunohistochemistry, the SALL4 gene was detected using reverse tran-scription-polymerase chain reaction (RT-PCR), and the relevant literature was reviewed. Results: Microscopically, all cases had primi-tive enteroid structures, consisting of cubic or columnar cells with clear cytoplasm, and immunohistochemical staining showed positivi-ty for either AFP and GPC3 or SALL4. The expression of SALL4 mRNA was significantly increased by RT-PCR. Follow-up from 1 to 5 years showed that 5 patients had liver and other organ metastases, 2 patients died, and 1 patient survived without a tumor. Conclusions:GCED is a rare invasive gastric adenocarcinoma with a worse prognosis than that of normal intestinal adenocarcinoma. The treatment of general intestinal adenocarcinoma has little effect. There are some characteristic changes in histology. It would be helpful for diag-nosis and differential diagnosis if clinicians are familiar with the tumor spectrum and genetic characteristics. Target therapy for an origi-nal marker, such as SALL4, has a bright future.

Objective To monitor the autophagy in osteosarcoma cells by constructing three rLC3B fusion expression vectors,respectively.Methods Rat LC3B gene sequence was amplified by PCR and cloned into pEGFP-C 1 and pmCherry-C1 to construct the fusion expression vector of pEGFP-rLC3B and pmCherry-rLC3B.Subsequently,the EGFP-rLC3B sequence was obtained by PCR with the pEGFP-rLC3B as a template,and cloned into pmCherry-C 1,so the pmCherry-EGFP-rLC3B fusion expression vector was constructed.Three plasmids were transfected into U-2OS cells,and the starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A1 was used to inhibit autophagy,to verify the above plasmids' function in autophagy detection by laser scanning confocal microscopy.Western blot was used to detect the endogenous LC3B and exogenous EGFPrLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B,and to verify the correct expression of exogenous rLC3B and their function of autophagy detection.Finally,cleaved free EGFP was detected by western blot to evaluate the level of autophagic degradation.Results Three fusion expression vectors were constructed successfully through sequencing and restriction enzyme digestion validation.The starvation or Rapamycin was adopted to induce autophagy or the chloroquine or Baf-A 1 was used to inhibit autophagy in transfected U-2OS cells.Clear autophagosomes and autolysosomes were observed by laser scanning confocal microscopy.Endogenous LC3B and exogenous EGFP-rLC3B,pmCherry-rLC3B and mCherry-EGFP-rLC3B were detected through western blot.Finally,western blot verified that the expression of cleaved free EGFP was significantly up-regulated with the increase of starvation time.12 h group increased 1.05 times than the control group and 24 h group increased 1.56 times,showing that the levels of autophagic degradation increased.Conclusion EGFP-rLC3B can be used to detect autophagosome and evaluate the level of autophagic degradation.mCherry-rLC3B can be used to detect autophagosome and autolysosome,but can't distinguish autophagosome from autolysosome.The pmCherry-EGFP-rLC3B has an advantage in the detection of autophagic flux which can distinguish autophagosome from autolysosome.

Objective To investigate the influence of isoquercitrin on the inflammatory factors in LPS-induced RAW264.7 cells.Methods MTT method was used to detect inhibition ratio of RAW264.7 cells induced by isoquercitrin.The level of TNF-α in culture medium was measured by ELISA.Nitric oxide (NO) was detected by Nitrate Assay Kit.Western blotting was used to investigate the influence on the productions of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2).Results The half inhibitory concentration (IC50) of isoquercitrin was 65.73 μmol·L-1.LPS had no inhibitory effect on the cells.Compared with LPS group,the level of TNF-α was decreased to 74.80% and 60.57% in isoquercitrin (20,10 μmol·L-1) groups in a dose-dependent manner.The results measured by Nitrate Assay Kit revealed that isoquercitrin (20,10 μmol·L-1) could suppress production of NO,the level of NO decreased to 79.34% and 68.81%(P<0.05).The Western blotting results showed that isoquercitrin (20,15,10 μmol·L-1) inhibited the productions of iNOS and COX-2 (P<0.05).Conclusion Isoquercitrin has anti-inflammatory effects by inhibiting the productions of TNF-α,NO,iNOS and COX-2,and the most effective dose for the inhibition is 10 μmol·L-1.

Aim To study the inhibitory effect of isoquercitrin on Raf/MEK/ERK signaling pathway in HepG2 cells.Methods MTT was used to detect the proliferation of human liver cancer HepG2 cells after the treatment of isoquercitrin.The morphology and growth of cells were observed under inverted microscope after the different concentrations of isoquercitrin(0, 40, 80, 160, 320 μmol·L-1) to treat HepG2 cells for 24 and 48 h.Cell cycle was assessed by flow cytometry.Ras, Raf, MEK, ERK expression was assayed by Western blot, and mRNA expression was detected by quantitative fluorescence PCR.Results Isoquercitrin could inhibit the growth of HepG2 cells in a concentration-and time-dependent manner.Typical morphological changes of apoptosis were observed by inverted microscopy after HepG2 cells were treated with different concentrations of of isoquercitrin for 24 h or 48 h.The cell cycle assay showed that with the increasing concentration of isoquerditrin, the number of cells that was arrested in G1 phase gradually increased.Compared with the blank group, the expressions of Ras, Raf, MEK, ERK mRNA were down-regulated, and related proteins expression were also down-regulated(P<0.05), and these results had statistical significance.Conclusion Isoquercitrin can induce the apoptosis of HepG2 cells, which may be related to the Raf/MEK/ERK signaling pathway.

Objective To investigate whetherthe extract of HUANGPI inhibitthe secretion of TNF-αvia TLR4/MyD88/TRAF6 signaling pathway. Methods ELISA assay was performed to determine TNF-α level in cell culture medium. MTT assay was used to detect the effects of the extract of HUANGPI and LPS on the viabilities of RAW 264.7 cells. Proteinexpressions of TLR4 and TRAF6 were detected by Western blotting assay. Results The extract of HUANGPI inhibited the secretion of TNF-αin a dose-dependent manner. Compared to LPS group , were TNF-αwas significantly suppressed in the cells in LPS+MyD88 inhibitor group , LPS+extract group and LPS+extract+MyD88 inhibitor group,with the corresponding reductions of TLR4 and TRAF6 protein expression at74% and21%,70% and27%,44% and8.5%, respectively. Conclusion MYD88-dependent signaling pathway might be involved in the mechanism underlying the effect of the extract of HUANGPI on suppressing LPS-induced inflammation.

Objective To evaluate the two-stage repair in hypospadias cripples.Methods 35 patients with hypospadias cripples underwent two-stage repair.At the first stage,a full-thickness graft of skin or buccal mucosa was used for urethral plate reconstruction after the release of chordee.The second stage was performed 6 months after completion of the first stage.Tubularization of the replaced urethral plate was the preferred technique.Results There was no urethra stricture or “preplaced urethral plate ” atrophy.Fistula developed in 2 of 35 cases.Conclusion The twostaged technique was a useful strategy to deal with the myriad abnormalities encountered in crippled hypospadias.This technique could not only create a neourethra successfully,but also give the penis a near-normal shape and appearance.

Objective To retrospectively analyze treatment status quo of giant cell tumor of bone around the knee in several institutes,and to investigate risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes.Methods A total of 222 patients with giant cell tumor of bone around the knee confirmed by pathology,who had undergone surgical treatment in 5 institutes from March 2000 to May 2012,were enrolled in this study.There were 120 males and 102 females,with an average age of 35.5 years.The epidemiology,clinical and radiographic features and risk factors affecting selection of surgical manners,tumor recurrence,and functional outcomes were analyzed.Results Intralesional curettage was performed in 128 patients(57.6％),intralesional curettage combined with partial resection in 13 patients (5.8％),and marginal excision in 79 patients (35.5％).A total of 159 patients were followed up.The local recurrence rate was 19.0％ for patients treated with intralesional curettage,8.9％ for marginal excision and 0％ for intralesional curettage combined with partial resection,and the total local recurrence rate was 14.5％(23/159).The mean duration from primary operation to recurrence was 23.9 months.Univariate analysis indicated that surgical manner was the only factor affecting local recurrence.Pathological fracture,Campanacci grades,Enneking system as well as soft tissue mass had a significant influence on the selection of surgical manners.The mean MSTS score after intralesional curettage was obviously higher than that after marginal excision.Conclusion The surgical manner is the only factor affecting local recurrence and limb function.