Objective: To study the local vascular remodeling, inflammatory response, and their correlations following acute spinal cord injury (SCI) with different grades, and to assess the histological changes in SCI rats. Methods: One hundred and sixteen adult female Sprague Dawley rats were randomly divided into 4 groups ( n=29). The rats in sham group were received laminectomy only. A standard MASCIS spinal cord compactor was applied with drop height of 12.5, 25.0, or 50.0 mm to establish the mild, moderate, or severe SCI model, respectively. Quantitative rat endothelial cell antigen 1 (RECA1) and CD68 positive areas and the correlations were studied by double immunofluorescent (DIF) staining at 12 hours, 24 hours, 3 days, 7 days, and 28 days following SCI. Moreover, qualitative neurofilament-H (NF-H) and glial fibrillary acidic protein (GFAP) positive glial cells were studied by DIF staining at 28 days. ELISA was used to detect the levels of tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), and IL-6 in spinal cord homogenates at 12 hours, 24 hours, and 3 days, and the correlations between TNF-α, IL-1β, or IL-6 levels and microvascular density (RECA1) were accordingly studied. Moreover, the neural tissue integrity and neuron damage were assessed by HE staining at 12 hours, 24 hours, 3 days, 7 days, and 28 days, and Nissl's staining at 28 days following SCI, respectively. Results: DIF staining revealed that the ratio of RECA1 positive area was the highest in moderate group, higher in mild and severe groups, and the lowest in sham group with significant differences between groups ( P<0.05). The ratio of CD68 positive area was the highest in severe group, higher in moderate and mild groups, and the lowest in sham group with significant differences between groups ( P<0.05), except the comparisons between mild and moderate groups at 24 hours and 28 days after SCI ( P>0.05). There was no significant correlation between the RECA1 and CD68 expressions in sham group at different time points ( P>0.05). At 12 and 24 hours after SCI, the RECA1 and CD68 expressions in mild and moderate groups showed significant positive correlations ( P<0.05), while no significant correlation was found in severe group ( P>0.05). No significant correlations between the RECA1 and CD68 expressions was shown in all SCI groups at 3 days and in severe group at 7 days ( P>0.05), while the negative correlations were shown in mild and moderate groups at 7 days, and in all SCI groups at 28 days ( P<0.05). In mild, moderate, and severe groups, the axons became disrupted, shorter and thicker rods-like, or even merged blocks with increased injury, while the astrocytes decreased in number, unorganized and condensed in appearance. ELISA studies showed that TNF-α, IL-1β, and IL-6 levels in sham group were significantly lower than those in other 3 groups at different time points ( P>0.05). The differences in TNF-α, IL-1β, and IL-6 levels between SCI groups at different time points were sinificant ( P<0.05), except IL-1β levels between the mild and moderate groups at 12 hours ( P>0.05). Three inflammatory factors were all significantly correlated with the microvascular density grades ( P<0.05). Histological analysis indicated that the damage to spinal cord tissue structure correlated with the extent of SCI. In severe group, local hemorrhage, edema, and infiltration of inflammatory cells were found the most drastic, the grey/white matter boundary was disappeared concurrently with the formation of cavity and shortage of normal neurons. Conclusion: In the acute stage following mild or moderate SCI, progressively aggravated injury result in higher microvessel density and increased inflammation. However, at the SCI region, the relation between microvessel density and inflammation inverse with time in the different grades of SCI. Accordingly, the destruction of neural structures positively relate to the grades of SCI and severity of inflammation.

Objective:To investigate the risk factors and interventions for surgical failure of spinal tuberculosis (STB).Methods:A total of 317 STB patients aged from 11 to 86 years with an average age of 53.5±16.7 years, who received debridement and fusion with bone grafting from January 2013 to December 2019, were retrospectively analyzed, including 206 males and 111 females. The follow-up duration was at least 1 year. During the follow-up, any one of the following 1)-3) was defined as surgical failure, namely 1) the same tubercular lesion treated by surgery more than 2 times, 2) the number of unplanned readmissions related to tubercular lesion≥1, 3) drug-resistant STB or delayed healing, recurrent lesion with cold abscess/sinus tract, combined with other bacterial infection, or loosening of internal fixation. The other cases were regarded as "curative" cases. Patients' symptoms, medication history, auxiliary examination and surgical plan were collected for univariate analysis. Further, the potential risk factors for surgical failure were analyzed by binary Logistic regression. Failed cases were treated with etiological intervention, such as puncture pumping pus or debridement or revision. The necrosis or granulation tissue was collected and further detected by tuberculosis culture, metagenomic next-generation sequencing (mNGS) and real-time fluorescent quantitative PCR (RT-qPCR).Results:There were 27 cases with surgical failure. Abscess or sinus tract formation was developed in 17 cases, which accounted for 63% (17/27). Among these patients, there were 3 cases of resistance to isoniazid or rifampicin and 2 cases of resistance to isoniazid and rifampicin (multidrug resistance, MDR). Seventeen cases were treated by anti-tuberculosis treatment, while 14 cases by puncture drainage (or puncture catheter irrigation) and 3 cases by debridement and suturing. Seven cases with wound infection or poor healing accounted for 26% (7/27). Among them, 5 kinds of pathogens were detected, none of which showed tuberculosis drug resistance. All of them were treated by anti-infection and debridement suturing, while 2 of them were treated with internal fixation removal. Three cases (11%, 3/27) with internal fixation loosening were treated by revision surgery. There was statistically significant difference between the failed group and the cured group in involved multi-/jumping segment, history of type 2 diabetes, a history of more than three basic diseases, CRP at one week after surgery, WBC at one week after surgery, time of first dose, operation duration and intraoperative blood loss ( P<0.10). Binary Logistic regression analysis showed that multi-/jumping segment ( OR= 3.513, P=0.047), CRP at one week after surgery ( OR=1.021, P=0.005), first dose time ≥20 weeks ( OR=2.895, P=0.039), blood loss ≥800 ml ( OR=5.950, P=0.001) and more than three basic diseases involved ( OR=3.671, P=0.027) were independent risk factors for surgical failure. Conclusion:Early diagnosis, especially the diagnosis of drug-resistant STB and standardized anti-tubercular treatment, should be carried out effectively. Puncture and drainage of abscess is an effective therapy to treat the cases with abscess/sinus tract formation. Some cases involved multi-/jumping segments could be with higher risk of failure after internal fixation. Thus, they should be treated individually with emphasis on the segmental stability reconstruction.

Objective:To explore the clinical value of peripheral remnant lipoproteins (RLP), low density lipoprotein cholesterol particle (LDL-P) and sdLDL particle (sdLDL-P) measurement in the diagnosis of carotid plaque, so as to provide practical basis for the accurate diagnosis of carotid plaque and the control of carotid plaque related cardiovascular and cerebrovascular diseases.Methods:People who underwent carotid plaque ultrasound examination in Xingtai Third Hospital , from January 2020 to June 2021 were selected as the research object. According to the ultrasound results, they were divided into carotid plaque group ( n=146) and control group without carotid plaque ( n=149). The fasting RLP, LDL-P and sdLDL-P of the two groups were measured by vertical auto profile (VAP) centrifugal separation phase, and the fasting TG and LDL-C were detected by routine mixed phase method. The indexes were compared between the two groups and the true positive rate, true negative rate, false positive rate and false negative rate of the diagnosis of carotid plaque were analyzed. The receiver operating characteristic curve of each test index was drawn, and AUC was used to evaluate the clinical diagnostic value of each test index for carotid plaque. Results:The levels of RLP, LDL-P and sdLDL-P in carotid plaque group were significantly higher than those in non-carotid plaque group ([1.07±0.36] mmol/L vs [0.59±0.17] mmol/L,[1 300±370] nmol/L vs [781±215] nmol/L,[435±139] nmol/L vs [156±59] nmol/L, all P<0.01). The true positive rate (78.08% [114/146],81.51% [119/146]) and true negative rate (84.56% [126/149], 86.58%[129/149]) of serum RLP and LDL-P for the diagnosis of carotid plaque were significantly higher than TG (58.90%[86/146], 43.62%[65/149]) and LDL-C (59.59% [87/146], 46.98% [70/149]), and the false positive rate (15.44% [23/149], 13.42% [20/149]) and false negative rate (21.92% [32/146], 18.49% [27/146]) were significantly lower than TG (56.38% [84/149], 41.10% [60/146]) and LDL-C (53.02% [79/149], 40.41% [59/146], all P<0.01). The AUC of the ROC curve of RLP (0.890), LDL-P (0.902) and sdLDL-P (0.973) for the diagnosis of carotid plaque was higher than TG (0.682) and LDL-C (0.712). The AUC of ROC curve of the RLP combined with sdLDL-P (0.977) for the diagnosis of carotid plaque was higher than the RLP and sdLDL-P (all P<0.01). Conclusion:The serum RLP, LDL-P and sdLDL-P can be used as indicators of carotid plaque, and their clinical diagnostic value are superior to TG and LDL-C; the combined diagnostic effect of lipoprotein subclass is better than that of single index alone.

Objective To investigate the clinical usefulness of quantitative dual-source dual-energy CT (DECT) iodine enhancement metrics combined with morphological CT features in distinguishing different lung cancer subtypes. Methods One hundred and sixty-two consecutive patients suspected with lung cancer were prospectively enrolled and underwent DECT in arterial phase prior to biopsy or surgery.Tumor histological subtypes were determined in 110 patients. Two radiologists interpreted CT morphologic features of 110 lesions in a consensual manner. In addition, two radiologists independently contoured lesions and placed regions of interest in descending aorta or subclavian artery on the same section for normalization , from which automated computer measurements were generated:iodine density and iodine ratio (the ratio of iodine density of lesion to that of artery on the same section). DECT metrics and morphological CT features were compared among different lung cancer subtypes. Chi-square was used to compare qualitative parameters. One way ANOVA was used to compare quantitative parameters satisfying normal distribution, while those parameters not satisfying normal distribution or ranked data were compared by Kruskal-Wallis rank sum test. Multinomial logistic regression models were used to differentiate the histological subtypes of lung cancer: adenocarcinoma, squamous cell carcinoma (SCC), small cell lung cancer (SCLC). Results There were 48 cases of adenocarcinomas, 36 cases of SCC and 26 cases of SCLC. In analysis of CT features, tumor diameter, distribution, spiculation, pleural retraction, vascular involvement, confluent mediastinal lymphadenopathy, encasement of mediastinal structures and enhancement heterogeneity showed statistical difference (all P<0.05). The diameter of SCC[(5.73 ± 3.67)cm] and SCLC [(6.08 ± 4.39)cm] were larger than adenocarcinoma [(3.75 ± 2.80 cm)] (H=13.806,P<0.05). Adenocarcinomas were mostly located in the periphery (31 cases), while SCC (26 cases) and SCLC (21 cases) were mainly centrally located. Spiculation was mostly found in adenocarcinoma (44 cases) rather than SCLC (13 cases). Pleural retraction was mostly observed in adenocarcinoma (36 cases) rather than SCC (10 cases) and SCLC (5 cases). Vascular involvement was mostly found in SCLC (19 cases) rather than adenocarcinoma (15 cases). Confluent mediastinal lymphadenopathy was more frequently found in SCLC (15 cases) compared with adenocarcinoma (3 cases) and SCC (4 cases). Encasement of mediastinal structures was mostly found in SCLC (13 cases) rather than adenocarcinoma (7 cases). Homogeneous enhancement was more frequently found in SCLC (10 cases) than SCC (6 cases). No significant differences were observed in other CT features between any other two groups. Iodine density and iodine ratio were statistically different among these three subtypes lung cancer (H=16.817,20.338,P<0.001). Iodine density of adenocarcinoma and SCC was (1.50±0.80) and (1.40± 0.40) mg/ml, respectively, higher than the (1.20±0.40) mg/ml for SCLC (P<0.01). Iodine ratio of adenocarcinoma and SCC was (16.10 ± 7.02)％and (15.05 ± 4.62)％, respectively, higher than the (11.55 ± 3.15)％ for SCLC (P<0.01). No significant difference was observed between adenocarcinoma and SCC. Accuracy of the model based on CT features was 69.1％, accuracy of the model based on CT features combined with DECT parameters was 80.9％. Conclusions Quantitative DECT metrics are different among adenocarcinoma, SCC and SCLC, when combined with morphological CT features, higher diagnostic accuracy can be achieved.

OBJECTIVETo investigate the effect of up-regulation of Rap1GAP on the invasion ability of leukemic HL-60 cells in vitro, and to establish leukemia mouse model to verify the effects in vivo.

METHODSQuantitative RT-PCR and Western blot methods were used to detect the expression of Rap1GAP in Venus/HL-60 (vehicle control) and Rap1GAP/HL-60 cells (R1 andR2). Transwell method was used to examine the invasion ability in vitro. Quantitative RT-PCR and gelatin zymograph were used to study the expression of MMP-2 and MMP-9. Four-week-old BALB/c nu/nu mice were pre-treated and inoculated with leukemic cells from different groups, several index including survival time were then monitored.

RESULTSRap1GAP mRNA level of R1 and R2 increased about 16-17 folds as compared to the control cells. The invasion rate of R1 and R2 are (55 ± 5)% and (59 ± 4)%, which are significantly higher than (14 ± 4)% of the control cells. The mRNA level of MMP-9 was up-regulated about 12.0 folds in R1 and R2 cells compared to the corresponding control cells. The median survival times of R1 and R2 mice are (32.00 ± 1.85) d and (33.37 ± 2.50) d, respectively, which are shorter than (43.62 ± 2.32) d of the control group. Three mice of R1 and R2 groups showed leukemic cells infiltration in meninges tissue, and the genes of Rap1GAP and MMP-9 were amplified by PCR method.

CONCLUSIONUp-regulated expression of Rap1GAP increased the invasion ability of HL-60 cells accompanied with enhancement of MMP-9 expression in vitro, and the experiment in mouse model also confirmed that Rap1GAP enhanced the invasion of HL-60 cells in vivo.

Animals ; GTPase-Activating Proteins ; metabolism ; HL-60 Cells ; Humans ; Leukemia ; Matrix Metalloproteinase 2 ; metabolism ; Matrix Metalloproteinase 9 ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Invasiveness ; RNA, Messenger ; Transcriptional Activation ; Up-Regulation

ObjectiveTo explore the effect and regulatory mechanism of notoginseng total saponins on apoptosis of mammary gland cells in rats with mammary gland hyperplasia. MethodSixty female non-pregnant SD rats were randomly divided into control group， model group， Notoginseng total saponins low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） groups and tamoxifen group （1.8 mg·kg^-1）， 10 rats per group. Rat model of mammary gland hyperplasia was established by intramuscular injection of estradiol benzoate and progesterone. Oral administration was performed according to the experimental dose of each group， once a day for 30 consecutive days. The rats in the control group and the model group were given equal volume of normal saline by gavage every day. After administration， the diameter of the second pair of nipples of the rats was measured with vernier calipers， and breast tissue samples were collected and stained with hematoxylin and eosin （HE） to observe the pathological changes. Immunohistochemistry was used to determine the expression of apoptosis regulators B-cell lymphoma-2 （Bcl-2）-associated X （Bax） and Bcl-2， and Western blot was conducted to detect the expression of phosphatidylinositol 3-kinase/protein kinase B/mammalian target of rapamycin （PI3K/Akt/mTOR） signaling pathway-related proteins. ResultCompared with the control group， the model group had increased diameter of the second pair of nipples （P<0.05）， elevated volume of mammary lobules and number of acinus， diffuse mammary gland hyperplasia， and up-regulated expression of Bcl-2 protein， increased ratio of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR （P<0.05） and decreased expression of Bax in the mammary gland （P<0.05）. Compared with the conditions in the model group， the diameter of the second pair of nipples of the rats in the notoginseng total saponins low-， medium- and high-dose groups and tamoxifen group was decreased （P<0.05）， and the number of mammary lobules and acinus and the amount of secretions were reduced. In addition， the mammary gland hyperplasia was alleviated， and a decrease was observed in the expression of Bcl-2 protein and the ratio of p-PI3K/PI3K， p-Akt/Akt， p-mTOR/mTOR （P<0.05）， and an increase in the expression of Bax （P<0.05）. ConclusionNotoginseng total saponins could improve mammary gland hyperplasia in rats， and its mechanism was related to regulating PI3K/Akt/mTOR pathway and promoting apoptosis of mammary gland cells.