Objective To evaluate the value of the periportal free air(PPFA) sign at computed tomography(CT) in distinguishing between upper and lower gastrointestinal(GI) tract perforation. Methods CT images of 62 patients with surgically proven GI tract perforation were retrospectively analyzed. 62 cases included upper and lower GI tract perforation in 35 cases and 27 cases,respective-ly. When there was free air in the periportal area,it was defined as positive periportal free air(PPFA) sign. The difference of PPFA sign in upper and lower GI tract perforation, and the sensitivity, specificity, positive predictive value, negative predictive value and ac-curacy of PPFA sign in diagnosing gastrointestinal perforation were analysed respectively. Results The PPFA sign was seen in 33 of 35(94%) patients with upper GI tract perforation,but only in 10 of 27 (37%) patients with lower GI tract perforation,there was sig-nificant difference between them(P<0.05). For diagnosis of super GI perforation with PPFA sign, the sensitivity, specificity, positive predictive value,negative predictive value and accuracy were 94% (33/35), 63%(17/27), 770%(33/43), 89%(17/19) and 81% (50/ 62), respectively. Conclusion The PPFA sign is a useful finding in distinguishing between upper and lower GI tract perforation.

Objective To evaluate the effect and potential mechanism of Shikonin on hormone-induced osteoporosis in rats.Methods RAW264.7 cell lines were treated with indicated concentrations of Shikonin,and the effects of Shikonin on cell viability were evaluated by CCK8 assay.Then the cells were divided into five groups:control group,dexamethasone(osteoporosis agent)group,dexamethasone + recombinant human parathyroid(anti-osteoporosis agent)group,dexamethasone + Shikonin 0.3 μmol·L-1 group,and dexamethasone + Shikonin 1.2 μmol·L-1 group.The effect of Shikonin on osteoclast function was evaluated by TRAP staining and bone resorption area measurement.The rat model of osteoporosis induced by dexamethasone was established,and divided into five groups(n=8):dexamethasone group,dexamethasone + PTH group,dexamethasone + Shikonin 0.3 μmol·L-1 group and dexamethasone + Shikonin 1.2 μmol·L-1 group,while control group was set up,then each group given separately 0.9%NaCl 6 mL·kg-1,PTH 20 μg·kg-1 3 days,Shikonin 0.5 mg·kg-1 day(0.3 μmol·L-1),Shikonin 2 mg·kg-1 day(1.2 μmol·L-1),and 0.9%NaCl 6 mL·kg-1for 3 weeks.ELISA was used to detect Serum type I collagen(CTX),Cathepsin K(Cathepsin K)and oxidative stress related indicators.HE staining was used to detect the changes of bone structure,and dual-energy X-ray scanning was used to detect the changes of bone density(BMD).Western blot and immunofluorescence were used to detect the effect of Shikonin on NF-κB/MAPKs signaling pathway,Western blot and qRT-PCR were used to detect the expression of RANKL-related proteins and mRNA,and Western blot was used to detect the blocking effect of Shikonin on RANKL/TRAF6.Results 1.2 μmol·L-1 Shikonin had the least damage to cells.Shikonin decreased the number of TRAP positive cells(P<0.05),decreased bone absorption area(P<0.05).and decreased the expression of CTX and Cathepsin K(P<0.05),and in 1.2 μmol·L-1 group decreased more significantly(P<0.05).Shikonin increased rat femur BMD(P<0.05),and increased more significantly in 1.2 μmol·L-1 group(P<0.01).Shikonin reversed the reduction and thinning of bone trabeculae induced by dexamethasone.The expression of SOD and GSH in rats increased(P<0.05),while the expression of MDA decreased(P<0.05).Shikonin inhibited the phosphorylation of NF-κB pathway related proteins in vitro.Shikonin inhibited the expression of RANK,RANKL,TRAF6,c-fos and NFATc1 protein and mRNA.Moreover,Shikonin blocked the expression of TRAF6.Conclusion Shikonin can ameliorate glucocorticoid induced osteoporosis by inhibiting RANKL/RANK/TRAF6 and its mediated NF-κB/MAPKs signaling pathway and oxidative stress.

Objective:To assess the value of amide proton transfer weighted (APTw) imaging in the evaluation of pH changes in infarct core (IC) and ischemic penumbra (IP) in subacute cerebral infarction.Methods:The data of twenty-three subacute cerebral infarction patients with unilateral steno-occlusive disease of the middle cerebral artery (subacute infarction group) from April to November 2019 in the First Affiliated Hospital of Dalian Medical University were prospectively analyzed. Fifteen healthy volunteers were enrolled in this study as the control group. All subjects underwent conventional MRI, DWI, 3D-pseudo continuous arterial spin labeling (3D-pCASL) and APTw sequences. Based on DWI images, relative cerebral blood flow (rCBF) and APTw images to determine the region of IC, blood flow penumbra [cerebral blood flow(CBF)-DWI mismatch area, IP CBF] and metabolic penumbra (APTw-DWI mismatched area, IP APT). 3D ROIs were used to semi-automatically measure the APTw signals and the volume of IC and IP CBF of the patients in subacute infarction group. The comparison of APTw signals between the infarct side and the contralateral side in the subacute infarction group, the comparison of bilateral APTw signals in the control group, and the comparison of APTw signals in the IC and IP CBF regions were performed by paired-sample t test or Wilcoxon signed-rank test. The paired-sample t test or Mann-Whitney U test was used to compare the APTw signals between the two groups. The Friedman test was applied to compare the difference of volumes among IP CBF1.5, IP CBF2.5 and IP APT . Results:There was no significant difference of the APTw signals among the IC, the contralateral side in the subacute infarction group and the control group ( P>0.05). The APTw signals of IP CBF and IC of the infarction group were statistically different ( P<0.05). Compared with the contralateral side of IP CBF1.5 (3.7±1.7, -1.84±1.48, 5.57±2.75), the APTwmax (3.07±1.41, t=-3.012, P=0.006), APTw min [-1.30 (-1.74, -0.57), Z=-2.099, P=0.036], and APTwmax-min(4.51±2.58, t=-3.273, P=0.003) signals in the IP CBF1.5 were decreased ( P<0.05). Compared with the contralateral side of IP CBF2.5 [-1.53 (-2.80, -0.91), 5.31±2.61], the APTw min [-1.08 (-1.60, -0.49), Z=-2.616, P=0.009] and APTwmax-min (4.41±2.72, t=-3.228, P=0.004) signals in the IP CBF2.5 were decreased. The volumes of IP CBF1.5 [107.51(50.08, 138.61)mm 3], IP APT [99.00 (53.27, 121.335) mm 3] and IP CBF2.5 [89.91 (51.53, 139.87) mm 3] were successively reduced (χ2=7.913, P=0.019), and the volume of IP CBF2.5 was significantly smaller than that of IP CBF1.5 ( P=0.037). Conclusion:The acid-base metabolism in the IC of subacute cerebral infarction is not obvious, but the blood flow penumbra has local acid-base metabolism imbalance, and the range of metabolic penumbra coincides with the blood flow penumbra.

Objective:To investigate and compare the clinical effectiveness and efficacy of cannulated screws and suture anchors in assistance of locking plates reinforcement for enhancing fixation in the treatment of proximal humeral fractures.Methods:A retrospective analysis was conducted on the data of 121 patients with proximal humeral fractures who were treated by locking plates reinforcement at Hainan Western Central Hospital from January 2018 to January 2020.Based on the intraoperative method of auxiliary reinforcement for the greater tuberosity of humerus,the patients were divided into cannulated screw group(62 cases)and suture anchor group(59 cases).The Disabilities of the Arm,Shoulder and Hand(DASH)score,Constant-Murley Shoulder Score and Neer Shoulder Function(Neer)Score pre surgery,the 3rd month and 12th month after surgery were recorded and compared between the two groups,so as to assess the changes in shoulder function.Results:The time of operation,the intraoperative bleeding volume,the length of stay and the follow up time of cannulated screw group were respectively(94.7±13.4)min,(230.5±39.2)ml,(9.2±1.3)d and(15.3±4.2)months,and the differences of them between two groups were no significant(P>0.05).All patients were satisfactory for the postoperatively proximal humeral fractures and the reduction of greater tuberosity,and there was no acromial impingement.The DASH scores of two groups at the 12th month after surgery were lower than those at the 3rd month after surgery,and the Constant-Murley scores and Neer scores of two groups at the 12th month after surgery were higher than those at the 3rd month after surgery,and the differences of them were significant(t=12.069,11.446,15.553,14.879,16.223,18.209,P<0.05),respectively.There were no statistically significant differences in DASH score,Constant-Murley score and Neer score at 3rd month and 12th month after surgery between the two groups(P>0.05).Conclusion:During surgical treatment for proximal humeral fractures,both cannulated screws and suture anchors can effectively assist to lock plate for strengthening fixation of the greater tuberosity,and maintaining anatomical reduction of the greater tuberosity,and promoting recovery of the function of shoulder joint.

Objective:To observe the effects of miR-21 knockout on proliferation and drug resistance in K562/G01 cells, and to preliminarily explore the mechanism of imatinib sensitivity by knocking out miR-21 in K562/G01 cells.Methods:Using CRISPR/Cas9 to knock out the miR-21 gene in K562/G01 cells, and single-cell-derived clones of miR-21 knockout were obtained by genomic DNA PCR screening, Sanger sequencing, and real-time PCR. We used MTT and cell colony formation assays to assess the cell proliferation, and determined imatinib sensitivity by MTT assay and Annexin-Ⅴ-APC/7-AAD double staining flow cytometry. Using western blot, we examined the potential mechanisms affecting imatinib sensitivity by knocking out miR-21 in K562/G01 cells.Results:Three miR-21 knockout K562/G01 single-cell-derived clones were successfully constructed. The mutation efficiency mediated by CRISPR/Cas9 was 7.12%-8.11%. MiR-21 knockout inhibited the proliferation of K562/G01 cells; the clone formation rates of WT and 1#, 2#, 6# K562/G01 single-cell clones were (57.67±8.25) %, (26.94± 5.36) %, (7.17±2.11) %, (31.50±3.65) %, respectively. MiR-21 knockout increased the sensitivity of K562/G01 cells to imatinib, IC 50 of imatinib in WT, and 1#, 2#, 6# K562/G01 single-cell clones were (21.92±1.36) μmol/ml, (3.98±0.39) μmol/ml, (5.38±1.01) μmol/ml, (9.24±1.36) μmol/ml. After the knockout of miR-21, the activation of PI3K/Akt signaling molecules was inhibited, while the expression of P210 BCR-ABL and p-P210 BCR-ABL was downregulated; however, the expression of PTEN was not affected. Conclusion:The knockout of miR-21 can suppress cell proliferation and improve sensitivity to imatinib in K562/G01 cells, which may be achieved by inhibiting the PI3K/AKT signaling pathway and BCR-ABL expression.

OBJECTIVE To analyze the chemical constituents from classical prescription Xiehuang San(XHS)standard decoc-tion by UPLC-Q-TOF-MS technology,and classify the chemical composition and analyze the representative components.METHODS Acquity HSS T3 column(2.1 mm×100 mm,1.8 μm)was used as the chromatographic column,with 0.1%formic acid solution-0.1%formic acid acetonitrile as the mobile phase for gradient elution.The volume flow rate was 0.4 mL·min-1 and the column tem-perature was 40℃.Mass spectrometry data of XHS were collected in positive and negative ion modes.The chemical constituents from classical prescription XHS were analyzed and identified by Masslynx 4.1 software comparison with reference materials,mass spectrome-try data analysis and reference to relevant literature.RESULTS A total of 107 compounds were analyzed and identified from XHS,including 45 flavonoids,27 triterpenoids,11 monoterpenoids,10 phenylpropanoids,6 chromogenic ketones,5 alkaloids and 3 other other compounds.CONCLUSION The study provides an experimental basis for the further research on the substance basis and qual-ity control of XHS.

Objective To investigate the effect of Trichomonas vaginalis macrophage migration inhibitory factor (TvMIF) on THP-1 macrophages.. Methods Recombinant TvMIF protein was prokaryotic expressed and purified, and endotoxin was removed after identification. Following exposure to TvMIF at concentrations of 0, 1, 5, 10, 50 and 100 ng/mL, the cytotoxicity of the recombinant TvMIF protein to THP-1 macrophages was tested using cell counting kit (CCK)-8 assay, and the apoptosis of THP-1 macrophages and reactive oxygen species (ROS) were detected using flow cytometry. The relative expression of nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3), caspase-1, interleukin-1β (IL-1β) and IL-18 genes was quantified using real-time fluorescent quantitative PCR (qPCR) assay, and the expression of caspase-1, NLRP3, gasdermin D (GSDMD), gasdermin D N-terminal (GSDMD-NT) and pro-IL-1β proteins were determined using Western blotting assay. Results Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) displayed successful expression and purification of the recombinant TvMIF protein with a molecular weight of 15.5 kDa, and the endotoxin activity assay showed the successful removal of endotoxin in the recombinant TvMIF protein (endotoxin concentration < 0.1 EU/mL), which was feasible for the subsequent studies on protein functions. Flow cytometry revealed that the recombinant TvMIF protein at a concentration of 10 ng/mL and less promoted the apoptosis of THP-1 macrophages, and the highest apoptotic rate of THP-1 macrophages was seen following exposure to the recombinant TvMIF protein at a concentration of 5 ng/mL, while the recombinant TvMIF protein at concentrations of 50 and100 ng/mL inhibited the apoptosis of THP-1 macrophages. Exposure to the recombinant TvMIF protein at a concentration 1 ng/mL resulted in increased ROS levels in THP-1 macrophages. qPCR assay quantified significantly elevated caspase-1, NLRP3, IL-18 and IL-1β expression in THP-1 macrophages 8 hours post-treatment with the recombinant TvMIF protein at a concentration 1 ng/mL, and Western blotting determined increased caspase-1, NLRP3, pro-IL-1β, GSDMD and GSDMD-NT protein expression in THP-1 macrophages following exposure to the recombinant TvMIF protein at a concentration 1 ng/mL. Pretreatment with MCC950 significantly reduced GSDMD and GSDMD-NT protein expression. Conclusions High-concentration recombinant TvMIF protein inhibits macrophage apoptosis, while low-concentration recombinant TvMIF protein activates NLRP3 inflammasome and promotes macrophage pyroptosis.