Objective To explore the mechanism of hepatitis B virus X protein down-regulating DKK4 and its effect on the proliferation, migration of HCC cell lines. Methods HCC cell lines HepG2 and SMMC7721 cells were infected with adenovirus encoding hepatitis B virus X protein (Ad-HBx), and GFP adenovirus (Ad-GFP) was designed as a control group. We used deacetylase inhibitor (TSA) to treat HCC cell lines and transfected HCC cell lines with small interfering RNA-histone deacetylase 1 (si-HDAC1) and lentivirus overexpressing DKK4. Western blot was used to detect the expression of DKK4, HDAC1 and SIRT1. The proliferation and migration ability of HCC lines were assessed using MTT, crystal violet experiment and Transwell experiment. Results DKK4 expression level was significantly downregulated after Ad-HBx infection (P < 0.05), and its expression level was recovered after TSA treatment (P < 0.05). After silencing HDAC1 with small interfering RNA, the expression of DKK4 could be restored (P < 0.05), the proliferation and migration of HDAC1-silencing or/and DKK4-overexpressing cells decreased (P < 0.05). Conclusion Hepatitis B virus X protein inhibits the expression of DKK4 protein by up-regulating HDAC1 and SIRT1. Silencing HDAC1 and over expressing DKK4 protein could inhibit the proliferation and migration of HCC cell lines infected with Ad-HBx.

Objective:To explore the expression level of exosomal miR-218-5p in the serum of patients with colorectal cancer (CRC) and its correlation with the clinical pathological characteristics, and evaluate its diagnostic efficacy in CRC.Methods:A group of 78 patients with colorectal cancer diagnosed in Tongji Hospital affiliated to Tongji University from October 2016 to October 2018 were selected. Blood was collected before operation and serum was preserved. Forty cases of healthy people in the same period were selected as the control group. ExoQuick kit was used to extract serum exosomes. Transmission electron microscope, NTA and western blot were used to identify the morphology and molecular phenotype of exosomes. MiRNeasy kit was used to extract total RNA in serum exosomes. Real-time fluorescent quantitative PCR (RT-qPCR) was used to detect the expression level of serum exosomal miR-218-5p in each group. CRC patients were divided into high expression and low expression groups using the median relative expression level of miR-218-5p as the cut off value, and the four-square chi-square test (χ 2 test) was used to judge the relationship between miR-218-5p and clinicopathological features. The receiver operating characteristic (ROC) curve was used to evaluate the diagnostic efficacy of miR-218-5p in colorectal cancer. Results:The exosomes in serum were successfully extracted by kit method. The expression level of serum exosomal miR-218-5p in patients with colorectal cancer was significantly lower than that in normal healthy people [0.566(0.364, 0.850) vs 1.054(0.781, 1.709), P<0.001]. The low expression level was significantly better then the correlated with tumor size, TNM stage, lymph node metastasis and depth of invasion (all P<0.05). Receiver operating characteristic (ROC) analysis showed that the area under the curve (AUC) of serum exosomal miR-218-5p for the diagnosis of CRC was 0.827 (95% CI 0.754-0.900), which was significantly better than the conventional tumor marker carcinoembryonic antigen CEA (AUC =0.718, 95% CI 0.626-0.811) and carbohydrate antigen CA199 (AUC = 0.661, 95% CI 0.564-0.758). Conclusions:The down-regulation of miR-218-5p expression in serum exosomes of colorectal cancer patients is associated with a variety of adverse clinicopathological factors, which has the potential to become a diagnostic biomarker for colorectal cancer.

Objective:To summarize the experience and the clinical data of patients with primary hyperparathyroidism undergoing endoscopic parathyroidectomy.Methods:A total of 24 patients who underwent endoscopic parathyroidectomy for primary hyperparathyroidism in Peking Union Medical College Hospital during Feb. 2021 to May. 2022 were concluded in this study (20 cases of parathyroidectomy via axillary approach and 4 cases of parathyroidectomy via thoracic and breast approach) . The operation time, postoperative drainage, length of stay, level of parathyroid hormone and serum calcium of those patients were collected. Postoperative complications and recurrence of hyperparathyroidism were also observed.Results:The postoperative levels of serum parathyroid hormone and serum calcium were significantly reduced (over 50%) compared with preoperative level ( P<0.05) . The average operation time was (96±22) min (64-157 min) . The mean postoperative drainage volume was (47±16) ml on day 1, (46±11) ml on day 2, and (30±9) ml on day 3, respectively. The average length of postoperative hospital stay was (2.8±1.1) days (2-6 days) . In one case of parathyroidectomy via axillary approach, the operation was converted to open surgery because of the low position of lesion. Other cases completed endoscopic surgery and obtained satisfactory cosmetic results. There were no postoperative complications such as bleeding, permanent hoarseness, coughing while drinking water, or surgical site infection. The mean follow-up time was (7.4±4.2) months (1-16 months) . There was no obvious discomfort and no recurrence during follow-up. Conclusion:Endoscopic parathyroidectomy is safe and effective in the treatment of primary hyperparathyroidism, which can be used as a surgical option for patients with cosmetic requirements.

Objective:To prepare pH-sensitive liposomes to avoid the degradation of monophosphoryl lipid A (MPLA) by lysosomes.Methods:Using DOPE and CHEMS as carrier materials, pH-sensitive liposomes were prepared by thin-film dispersion method. Particle sizes and Zeta potential of the liposomes were detected by dynamic light scattering. The morphological features of pH-sensitive liposomes under different pH conditions were observed by transmission electron microscopy. Flow cytometry was performed to detect the phagocytosis of liposomes by THP-1 and DC2.4 cells. Confocal laser microscopy was used to observed the colocalization of liposomes and lysosomes. BALB/c mice were immunized with hepatitis B surface antigen (HBsAg) using MPLA pH-sensitive liposome as an adjuvant. The levels of serum anti-HBs were quantitatively detected by ELISA. IFN-γ and IL-2 spot forming cells (SFCs) in mouse splenic lymphocytes were detected by ELISPOT.Results:The pH-sensitive liposomes were constructed with an average particle size of (90.90±1.13) nm, polydispersity index (PDI) of 0.076±0.013 and Zeta potential of (-27.900±0.666) mV. As the pH value of the solution decreased, the particle size increased significantly and the liposomes presented irregular shapes, indicating the pH-sensitive features. The phagocytosis rates by THP-1 cells and DC2.4 cells were 10.40% and 12.40% for pH-sensitive fluorescent liposomes, and 1.09% and 0.28% for fluorescent liposomes. Confocal laser microscopy revealed that pH-sensitive fluorescent liposomes were phagocytosed by THP-1 cells and existed in the cytoplasm, while fluorescent liposomes existed in lysosomes. Compared with MPLA liposomes, MPLA pH-sensitive liposomes could significantly improve the cellular immune response in mice. The levels of IFN-γ and IL-2 SFCs in the MPLA pH-sensitive liposome group were significantly higher than those in the MPLA liposome group ( P<0.01) and the non-adjuvant group ( P<0.001). Conclusions:The pH-sensitive liposome delivery system could improve the utilization of MPLA as an adjuvant.

Objective To investigate the correlation of spinal mechanical imbalance and thoraco-dorsal pain of ankylosing spondylitis (AS). Methods The clinical data of 90 patients with AS were collected. Patients were divided into two groups according to the presence of thoracodorsal pain: the AS with thoraco-dorsal pain group (30 cases) and the AS without thoraco-dorsal pain group (60 cases). Clinical symptoms, Bath ankylosing spondylitis disease activity index (BASDAI), Bath ankylosing spondylitis function index (BASFI), Bath ankylosing spondylitis measurement index (BASMI), ankylosing spondylitis disease activity (ASDAS), and spinal mechanical function and nuclear myocardial force test were compared using t-test, one-way analysis of variance (ANOVA) analysis and Spearman correlation analysis. Results ① There were differences between thoraco-dorsal pain group and patients without thoracodorsal pain group at the time of back muscle strength [(0.82±0.41) min vs (1.33±0.74) min, F=12.372, P=0.001]; ②Thoraco-dorsal pain in the AS group was mainly the middle and lower thoracic vertebrae, such as the inflammation of rib head and rib transverse process, facial arthritis, and spinous ligaments, etc. And the missed diagnosis rate of magnetic resonance imagin (MRI) was high. ③ In healthy control group, the anterior flexion strength of thoracodorsal pain group was signific-antly different from that of patients without thoracodorsal pain [(92.1 ±46.3) Nm vs (126.6±35.7) Nm, F=6.440, P=0.002]. ④ There was significant difference in spinal strength as well as left and right rotation strength between the thoracodorsal pain group and patients without thoracodorsal pain [(1.18 ±0.22) vs (1.05 ±0.17), F=10.044, P<0.01];⑤In the thoraco-dorsal pain group, the right/left index was related to BASDAI (r=-0.522, P=0.004). For spinal mobility, the right/left index was related to cross cutting faces to right ( r=0.435, P=0.021), cross cutting faces to left (r=0.528, P=0.004). In spinal strength, the right/left index was related to left turn (r=0.57, P=0.001); right lateral flexion (r=0.368, P=0.049) and left lateral flexion (r=0.369, P=0.049). Conclusion The thoracodorsal pain of AS is dominated by the middle and lower thoracic vertebrae, and the missed diagnosis rate of MRI is high. The imbalance of the left and right side of the spine is one of the factors of the thoracic back pain in AS.

Screening and evaluating the diseases responding specifically to traditional Chinese medicine （TCM） will help to highlight the advantages of TCM treatment， and the evaluation method should be standardized with consideration to the unique characteristics of the diseases. The incidence of Sjögren's Syndrome （SS） is increasing year by year， while the pathogenesis of this disease remains unclear. Modern therapies for this disease include biological agents and immunosuppressants， which generally have unsatisfactory efficacy. The TCM treatment of SS focuses on the harmony of the physical and mental health. The Rheumatology Branch of the China Association of Chinese Medicine organizes experts in TCM， Western medicine， and evidence-based medicine to form working groups. Delphi method and bibliometric method were used for analysis， and SS was selected as a disease responding specifically to TCM. Furthermore， the evaluation system was established for this disease， and the consensus regarding this disease was reached after seminar discussion. This paper summarized the whole process of the evaluation of the advantages of TCM treatment of SS. First， because TCM atomization is widely used in clinical practice and enriches TCM administration methods， this therapy is included after other non-drug therapies were taken as characteristic therapies. Second， the evaluation indicators of therapeutic effect should be determined with consideration to international acceptance and the current research status. Third， the expression method should be accurate， standardized， and objective， highlight the natural advantages of TCM， and avoid arbitrary extension. This paper provides a reference for clinicians to explore other diseases responding specifically to TCM.