OBJECTIVE To explore the mechanism of limonin treating in hepatic fibrosis through network pharmacology, and validate its mechanism by molecular docking and animal experiments. METHODS Firstly, the targets of limonin and hepatic fibrosis were screened from the SwissTargetPrediction, GeneCards and DisGeNet database, etc. Meanwhile, the common targets of limonin and hepatic fibrosis were obtained from the bioinformatics website. The protein protein interaction network of common target was constructed by using STRING database and Cytoscape software, and the CytoNCA plug-in was used to screen core targets. And then the enrichment analysis of GO and KEGG on the common target was performed by Metascape database. Thereby, the possible mechanism of limonin against hepatic fibrosis were predicted. Finally, the AutoDock Vina was used for molecular docking verification, and the prediction results of network pharmacology were verified by animal experiments. RESULTS The prediction results indicated that limonin might acted on 86 targets including AKT1, VEGFA and HIF1A, and participated in biological processes including hormone response, protein phosphorylation, angiogenesis, and PI3K-Akt pathway, HIF-1 pathway, VEGF pathway and other signaling pathways related to hepatic fibrosis. The results of protein protein interaction network topology analysis showed that the 11 core targets including AKT1, VEGFA, HIF1A and PIK3CA, etc. Molecular docking results showed that limonin had strong affinity and relatively stable binding conformation with the core targets. In the animal experiments, compared with the model group, hyaluronidase(HA) and laminin(LN) in rat serume in high-dose group of limonin(LH) and low-dose group of limonin(LL)(except for LN in LL group) were declined(P<0.01 or P<0.05), and the degree of inflammation and hepatic fibrosis were relieved to different degrees in liver tissue of the LH group and LL group; Western blotting and qPCR detection showed that protein and mRNA expression levels of AKT, HIF-1α and VEGF(except for VEGF in LL group) was down-regulated in the LH group and LL group(P<0.01 or P<0.05). CONCLUSION Limonin may acts on AKT1, VEGFA, HIF1A and other core targets to treat hepatic fibrosis angiogenesis, which may be related to the inhibition of AKT/HIF-1α/VEGF signaling pathway.

A three-dimensional feature construction method based on spectral information is presented,in which the power values of each channel are arranged into two-dimensional feature vectors based on electrode positions.The different frequency band features are arranged into a three-dimensional integral feature tensor to extract the information in frequency domain.Meanwhile,in order to reduce the influence of volume conductor effect,functional connectivity is utilized to map the temporal electroencephalogram data to the spatial brain functional network for extracting the spatial information.By analyzing the relationship between features and target classes,a 3D-CNN-Attention network model is proposed to incorporate an Attention mechanism in 3D-CNN network to enhance the electroencephalogram feature learning capability.A series of comparative experiments on publicly available datasets show that 3D-CNN-Attention network framework outperforms other methods in depression detection,obtaining an accuracy rate of up to 96.32%.The proposed method provides an effective solution for depression detection.

Objective:To investigate the correlation between paraspinal muscle atrophy, morphological changes of facet joints and adjacent segment disease (ASDis) after lumbar fusion operation.Methods:A retrospective study was conducted among 195 patients who underwent posterior lumbar fusion again for ASDis at this institution from January 2014 to December 2020, including 29 patients with ASDis whose initial surgical fusion segment was L 4,5. According to Roussouly's staging, there were 5 cases of type I, 9 cases of type II, 10 cases of type III, and 5 cases of type IV. Another 29 cases were selected from patients without ASDis after lumbar fusion as a control group. The control group was paired 1∶1 with the ASDis group according to gender, fusion segment, and Roussouly typing of the lumbar spine. The cross-sectional area (CSA) and fat infiltration (FI) of paravertebral muscle, facet joint angle (F-J) and pedicle facet (P-F) angle before the first (second) operation were measured and compared between the two groups. Then logistic regression analysis was used to determine the predictors of ASDis after posterior lumbar fusion. Finally, the receiver operation characteristic (ROC) curve was described, and the area under the curve (AUC) and cut-off point were calculated. At the same time, the paraspinal muscle atrophy before the second operation in ASDis group was measured. Results:The average follow-up time of 98 patients was 59.25±6.38 months (range, 49-73 months). The average body mass index (BMI) of ASDis group was 24.76±3.64 kg/m 2, which was higher than that in control group (22.24±2.92 kg/m 2) ( t=2.481, P=0.041). The average CSA and relative cross-sectional area (rCSA) of paraspinal muscle in ASDis group were 3 214.32± 421.15 mm 2 and 1.69±0.36 respectively, which were less than 3 978.91±459.87 mm 2 and 2.26±0.29 in control group ( t=10.22, P=0.012; t=9.47, P=0.038). The FI degree of paraspinal muscle in ASDis group (21.95%±5.89%) was significantly higher than that in control group (14.64%±7.11%) ( t=7.32, P=0.002). The F-J angle in ASDis group was 35.06°±3.45°, which was less than 38.39°±4.67° in control group ( t=4.76, P=0.027). The P-F angle in ASDis group was 117.39°±8.13°, which was greater than 111.32°±4.78° in control group ( t=5.25, P=0.031). Multivariate logistic regression analysis showed that higher BMI ( OR=1.34, P=0.038), smaller rCSA of paraspinal muscle ( OR=0.02, P=0.017) and higher FI of paraspinal muscle ( OR=1.58, P=0.032) were the risk factors of postoperative ASDis. The ROC curve showed that the AUC of BMI was 0.680 and the cut-off point was 22.58 kg/m 2; The AUC of the FI of paraspinal muscle was 0.716 and the cut-off point was 15.69%; The AUC of rCSA of paraspinal muscle was 0.227 and the cut-off point was 1.92. For ASDis patients, the paraspinal muscle before the second operation had a higher degree of FI (25.47%±6.59% vs. 21.95%±5.89%, t=3.99, P=0.042) and a smaller rCSA (1.52±0.28 vs. 1.69±0.36, t=3.85, P=0.038) than that before the first operation. The difference between the FI degree of paraspinal muscle before the second operation and the first operation was negatively correlated with the occurrence time of ASDis ( r=-0.53, P=0.039) , and the difference of rCSA was positively correlated with the occurrence time of ASDis ( r=0.64, P=0.043) . Conclusion:When BMI >22.58 kg/m 2, FI of paraspinal muscle >15.69%, and rCSA of paraspinal muscle <1.92, it suggests that ASDis is more likely to occur after operation. And the more obvious paraspinal muscle atrophy after the first operation, the earlier ASDis may occur. Morphological changes of facet joints cannot be used as an index to predict the occurrence of ASDis.

Protein self-assemblies at the micro- and nano-scale are of great interest because of their morphological diversity and good biocompatibility. High-throughput screening of protein self-assembly at different scales and morphologies using protein crystallization screening conditions is an emerging method. When using this method to screen protein self-assembly conditions, some apparently transparent droplets are often observed, in which it is not clear whether self-assembly occurs. We explored the interaction between β-lactoglobulin and the protein crystallization kit Index™ C10 and observed the presence of micro- and nano-scale protein self-assemblies in the transparent droplets. The diverse morphology of the micro- and nano-scale self-assemblies in the transparent droplets formed by mixing different initial concentrations of β-lactoglobulin and Index™ C10 was further investigated by scanning electron microscope. Self-assembly process of fluorescence-labelled β-lactoglobulin was monitored continuously by laser confocal microscope, allowing real-time observation of the liquid-liquid phase separation phenomenon and the morphology of the final self-assemblies. The internal structure of the self-assemblies was gradually ordered over time by in-situ X-ray diffraction. This indicates that the self-assembly phenomenon within transparent droplets, observed in protein self-assembly condition screening experiments, is worthy of further in-depth exploration.
Crystallization ; Lactoglobulins

[Abstract] Objective: To investigate the expression of chemokine-like factor-like MARVEL transmembrane domain-containing family member 6 (CMTM6) in breast cancer tissues and its correlation with clinicopathological features and prognosis of patients. Methods:Atotal of 136 breast cancer tissue chips (purchased from Superchip Company), including 42 pairs of matched cancer and paracancerous tissues, were used for this study. The expression level of CMTM6 in cancer and paracancerous tissues was detected by immunohistochemistry. The comparison of CMTM6 expression between breast cancer and paracancerous tissues was conducted by paired χ2 test. The relationship between CMTM6 expression in breast cancer tissues and the clinicopathological characteristics of patients was analyzed by χ2 test. Kaplan-Meier and Log rank test analyses were used to analyze the relationship between CMTM6 expression and the survival of patients, and Cox model was used to evaluate the effect of different indicators on the prognosis of patients. Results: The expression of CMTM6 in breast cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The expression of CMTM6 was correlated with pathological type of breast cancer and HER2 positivity (P<0.05). The survival time of patients in CMTM6 high expression group was significantly shorter than that of patients in CMTM6 low expression group (P<0.05). Pathological type (HR=10.374, 95%CI: 3.529-30.497, P<0.01), TNM stage (HR=4.599, 95%CI: 1.784-11.856, P<0.01), triple-negative breast cancer (HR=3.370, 95%CI: 1.055-10.761, P<0.05) and high expression of CMTM6 (HR=0.195, 95%CI: 0.073-0.518, P<0.01) were independent risk factors for prognosis of breast cancer patients. Conclusion: CMTM6 is highly expressed in breast cancer tissues, which can be used as a risk factor for prognosis evaluation of breast cancer patients.

Objective: To compare the efficacy and safety of plasma exchange (PE) combined with double plasma absorption and simple PE in the treatment of acute-on-chronic liver failure. Methods: We retrospectively analyzed 251 cases of acute-on-chronic liver failure treated with artificial liver treatment since January 2015. Changes in clinical manifestations, laboratory tests, and complications of the patients before and after different modes of treatment were compared and short-term efficacy was tracked. In accordance with different data, t-test, Pearson's chi-squared test and Fisher's exact test were used for statistical analysis. Results: The effectiveness of low-volume PE combined with double plasma molecular adsorption system (DPMAS) and equal amount of PE combined with DPMAS was significantly better than simple PE (83.7%, 84.05% and 82.15 vs 55.6%, P < 0.05) in early stage of liver failure. In late-stage of liver failure, there was no significant difference in the treatment efficiency of each group (P > 0.05). Bilirubin and bile acid levels were significantly decreased in combined treatment groups than that to simple PE group (P < 0.05). PTA and albumin improvement rate of DPMAS PE groups were significantly lower than that of simple PE group (P < 0.05). There was no statistical difference in adverse reactions between each group. Conclusion PE combined with DPMAS improves the treatment efficiency of early hepatic failure and decrease dosage of plasma when compared with simple PE. A beforehand DPMAS treatment after PE treatment can improve the adverse effects of DPMAS on blood coagulation function and albumin levels.

Objective To study the clinical effect of total parathyroidectomy (TPTX) on treatment of secondary hyperparathyroidism (SHPT) in patients with chronic renal failure.Methods 24 patients undergoing TPTX in Mianyang Central Hospital from Jan.2014 to Nov.2016 were retrospectively analyzed.The preoperative,postoperative and follow-up intact parathyroid hormone (iPTH),serum calcium,serum phosphorus and calciumphosphorus product were statistically analyzed.The postoperative complications and recurrence were observed.Results For preoperative,1 week after operation,3 months after operation,6 months after operation,12 months after operation,24 months after operation and 36 months after operation,the mean values of iPTH were (1771.7± 244.5) pg/ml,(70.2±18.2) pg/ml,(106.5±35.3) pg/ml,(114.3±31.3) pg/ml,(122.1±27.8) pg/ml,(128.1±12.5) pg/ml and (113.8±24.9) pg/ml.The mean values of serum calcium were (2.6±0.4) mmol/L,(1.9±0.4) mmol/L,(2.1±0.3) mmol/L,(2.1±0.2) mmol/L,(1.8±0.2) mmol/L,(1.8±0.2) mmol/L and (1.8±0.3) mmol/L.The mean values of serum phosphorus were (2.1±0.7) mmol/L,(1.4±0.6) mmol/L,(1.3±0.4) mmol/L,(1.4±0.3) mmol/L,(1.3±0.3) mmol/L,(1.5±0.2) mmol/L and (1.4±0.4) mmol/L.The mean values of calcium-phosphorus product were (61.9±17.0) mg2/dl2,(34.6±19.4) mg2/dl2,(37.4±14.4) mg2/dl2,(40.4±12.3) mg2/dl2,(29.2±5.3) mg2/dl2,(35.0 ±7.1) mg2/dl2 and (32.4 ± 11.3) mg2/dl2,respectively.Compared with those before operation,postoperative iPTH,serum calcium,serum phosphorus and calcium-phosphorus product decreased significantly at 1 week after operation,3 months after operation,6 months after operation,12 months after operation,24 months after operation and 36 months after operation,and the differences had statistical significance (all P＜0.05).Temporary injury of recurrent laryngeal nerve was found in one patient (4.2％).Early postoperative hypocalcemia rate was 91.7％ (22/24).No recurrence developed after operation.Conclusion TPTX is safe and effective in treatment of SHPT in patients with chronic renal failure,with low recurrence rate and satisfactory treatment outcome.

The main work and achievements of clinical pathway work in China since 2009 were systematically reviewed in the paper. It analyzed the problems existing in the implementation of clinical pathway management in China, and suggested on such management in the future. The suggestions include:deeper understanding, convergence with the payment system reform, strengthened quality control, further informatization,and better performance appraisal system.

Objective To prospectively summary the piercing-out position,direction,length and piercing-in position of perforator,and investigate the feasibility of preoperative design and optimization of the anterolateral thigh flap and its clinical application.Methods All 58 cases of anterolateral thigh flaps were designed and taken from the lateral thigh area from January,2014 to January,2016.Portable Doppler ultrasound was used before an operation to detect the piercing-out position (point P) of perforators.The direction and length (lower subcutaneous segment of perforators) of perforators after leaving piercing-out position were observed during the operation.And the piercing-in positions (point P') on superficial fascia and the dermis were observed.Based on this,we added line B (anterior superior spine-lateral femoral epicondyle) and line C (anterior superior spine-the middle point of superior border of patella) in the lateral and anterior side of original ilium-patella line in the thigh (line A),respectively.Results All perforators found in 58 cases before and during the operations were located on line A or between line A and line B.No perforators were found between line A and line C.Perforators walked toward the anterior medial side after leaving the muscle membrane.The perforator vascular subcutaneous segment (distance between point P and point P') was (2.02±0.23) cm.There was rectus muscle branch in the descending branch of lateral femoral circumflex artery,while no rectus muscle cutaneous branch was seen.20 cases were designed by one-line method,12 cases were designed by two-line method,while 26 cases were designed by three-line method.Conclusion Advanced three-line method is beneficial to detect of the perforators on the anterior thigh lateral region and to reduce the intraoperative injury perforator vessels at the puncture point.Clinical application of the anterior lateral thigh flap is simple and reliable.