Objective:To isolate and culture the pulp cells from human young permanent teeth (pDPC),and to observe their biological characteristics and the expression of some specific markers,and to induce these pulp cells to differentiate into osteoblast, adipocyte, neuron and chondrocyte lineages. Methods:Pulp cells were isolated and cultured from orthodontic extracted premolars of children. The attached cells after at least 3 passages were used for the following experiments:1. Morphology and ultrastructure analysis; 2. Cell cycle and phenotype were analyzed by flowcytometry; 3. Growth curve were recorded;4. pDPC were induced to differentiate into osteoblast,adipocyte,neuron in vitro,and were identified by histochemical methods and RT-PCR. Results: 1. Attached pDPCs were fibrablast-like cells,which were distinguished from BMSC. 2. The cell organs in dDPCs were well developed. 3. pDPCs were highly positive for CD90, CD44, CD147,which are mesenchymal stem-cell markers,but were negative for other markers including CD34, CD38, CD45, HLA-DR. 4. pDPCs showed high growth rate. 5. pDPCs could be induced to differentiate into osteoblast, adipocyte, and neuron lineages,but not chondrocyte lineages. Conclusion:pDPCs were characterized by their ability to proliferate with high growth rate in vitro. The expression of some BMSC markers in these cells were observed. They showed the potential to differentiate into multiple mesenchymal lineages such as osteoblst,adipocyte, neuron lineages under specific conditions in vitro.

Objective To knock out p21 gene in human malignant rhab doid tumor(MRT)cell line G401 by using CRISPR/Cas9 genome engineering technology. Methods The expression of p21 was detected by reverse transcription quantitative PCR (RT-qPCR) and Western blot assay in several MRT cell lines. The guide RNA was designed by targeting the third exon of p21 gene,which encoded its home domains, and then subcloned into lentiCRISPR v2 vector and validated sequencing. The validated plasmids were further used to package and produce the lentivirus in 293T cells, and the G401 cells were infected, then puromycin was used to screen positive cells, and the clusters of G401 monoclonal cells, were obtained by selecting monoclonal cells and culturing under the microscope. The RNA and protein of new clonal cell line were extracted, and RT-qPCR and Western blot assay were applied to confirm whether p21 was successfully knocked out. Results The p21 was highly expressed in MRT tumor cells. The CRISPR/Cas9 lentivirus plasmids, targeted p21 gene were successfully constructed. Compared with negative control group,the expression of p21 was not detected in G401 monoclonal cells, which were successfully screened. Conclusion In view of the difficult transfection of cells such as G401, p21 knockout stable cell line has been successfully constructed by using CRISPR/Cas9 system, which lays the foundation for further study of the mechanism of p21 in MRT tumors .

OBJECTIVE:To study the improvement effects of Rouganbao granules on liver fibrosis of model rats,and to provide experimental evidence for further development of this preparation. METHODS:Totally 45 male SD rats were divided into normal group,model group,positive control group(Fuzheng huayu capsules,0.4 g/kg)and Rouganbao granules high-dose and low-dose groups(16.8,8.4 g/kg by crude drug)according to random number table,with 9 rats in each group. Except for normal group,other groups were given CCl4sub cutaneously and high lipid diet+15% ethanol solution for 12 weeks to establish liver fibrosis model. After modeling,medication groups were given relevant medicine intragastrically,once a day,for consecutive 12 weeks. Normal group and model group were given corresponding volume of water intragastrically. The pathology change of liver tissues of rats was observed by inverted microscope. The levels of liver function indexes (ALT,AST,ALB) and liver fibrosis indexes(HA,Ⅳ-C,LN,Ⅲ-PC)were determined by ELISA. RESULTS:Compared with model group,liver cell vacuolation and liver fibrosis of rats were improved significantly in Rouganbao granule high-dose and low-dose groups;central vein was clearly visible,and no dilation and atrophy were found,especially in high-dose group;the serum levels of ALT and AST were decreased significantly,while the level of ALB was increased significantly(P＜0.05 or P＜0.01),of which the protective effect for liver was similar to positive control drug but better in increasing ALB(P＜0.05 or P＜0.01);the serum levels of HA,Ⅳ-C,LN and Ⅲ-PC were all decreased in varying degree,especially in high-dose group(P＜0.05 or P＜0.01),of which the effect of relieving liver fibrosis was similar to positive control drug. CONCLUSIONS:Rouganbao granules can improve liver function and delay the progression of liver fibrosis in model rats.

Objective:To evaluate the application of metagenomic next-generation sequencing (mNGS) in the diagnosis of osteoarticular infection.Methods:The clinical data of 37 inpatients aged 32-90 year with osteoarticular infection admitted in the Department of Spine Surgery of Qingdao Chest Hospital from January to December 2019 were retrospectively analyzed. There were 31 cases of spine infection and 6 cases of other joint infection. The tissue samples were obtained from the infected sites through puncture or surgical approach in all patients. The tissue samples were subjected to routine culture of mycobacteria, aerobic bacteria and anaerobic bacteria, respectively. The gene amplification and mNGS were performed for detection of mycobacterium tuberculosis DNA (MTB-DNA). The chi-square test or Fisher’s exact test were used to compare the detection rates of pathogen and simple bacterial infection between mNGS and conventional culture. The conventional culture, mNGS and MTB-DNA amplification detection were performed for all samples; with clinical diagnosis as the gold standard, the diagnostic values of 3 methods were evaluated with receiver operating characteristic curve (ROC). Paired sample t test was used to compare white blood cell(WBC) count, erythrocyte sedimentation rate, C-reactive protein of patients before and after treatment. P<0.05 was considered statistically significant. Results:The pathogens were detected by mNGS for 42 times: bacteria for 39 times (92.8%), fungi for twice (4.8%) and Kirks body for once (2.4%). Among 37 patients there were 29 cases of pure bacterial infection (78.4%), 2 cases of pure fungi infection (5.4%), 1 case of pure Kirks body infection (2.7%), and 5 cases of mixed infection of two or more pathogens (13.5%). The detection rates of mNGS and conventional culture were 100.0% (37/37) and 67.6% (25/37), respectively ( χ2=13.987, P<0.05). The detection rates of mNGS and conventional culture in 29 patients with pure bacterial infection were 100.0% (29/29) and 69.0% (20/29), respectively ( χ2=16.913, P<0.05). The area under the ROC curve (AUC) of conventional culture, mNGS, and MTB-DNA in the diagnosis of osteoarticular tuberculosis infection was 0.958 (95% CI: 0.866-1.000, P<0.05), 1.000 (95% CI: 1.000-1.000, P<0.05) and 0.958 (95% CI: 0.866-1.000, P<0.05). All the 37 patients were treated with anti-infective drugs according to the results of mNGS and conventional culture. Among them, 28 patients received surgical intervention. The patients were followed up until April 30, 2020, 1 patient died. After 3 months of follow-up, the WBC count, erythrocyte sedimentation rate and C-reactive protein were (5.5±1.5)×10 9/L, (41±38)mm/h and (5.0±4.6) mg/L, respectively, which were lower than those before anti-infection treatment [(8.0±2.9)×10 9/L, (79±42)mm/h and(63±52)mg/L] ( t=6.536, 8.302 and 6.373, all P<0.05). Conclusion:The metagenomic next-generation sequencing may have important clinical value in the differential diagnosis of osteoarticular infection.

Objective:To summarize the experience of five hospitals in Guangdong Province on successfully transporting retained passengers by chartered flights during the outbreak of the COVID-19.Methods:We retrospectively evaluated the operation and management, cabin arrangement, isolation requirements, and personnel protection in the aspect of epidemic prevention and control.Results:In 11 charter missions, over one thousand "healthy" passengers with potential risk of infections were transported back to China. The medical delivery team and flight crew were kept free of infection, passengers maintained zero cluster infections and no unexpected adverse events during the air transport.Conclusions:Our results indicate how to carry passengers in a scientific and orderly way is crucial for avoiding the transmission risks of the epidemic of COVID-19 among working staffers and passengers during air transportation.

Objective To investigate the influencing factors of opioid analgesics need within 48 hours after arthroscopic rotator cuff repair.Methods Clinical data of 90 consecutive arthroscopic rotator cuff repairs by the same operator from March 2017 to July 2022 were retrospectively analyzed.The patients were divided into opioid group(62 patients)and control group(28 patients)according to whether they used opioid analgesics within 48 hours after surgery.The control group consisted of patients who did not use analgesics or who had good analgesic effects with conventional analgesic regimens(non-steroidal anti-inflammatory drugs or non-opioid central analgesics)after surgery,and the opioid group consisted of patients who required opioid analgesics as assessed by the surgeon or who need opioid analgesics because of inefficacy of conventional analgesic regimens.The following factors were selected as independent variables:gender,age(whether≥65 years old),duration of disease(whether≥4 weeks),body mass index(BMI)(whether≥25),occupation(whether employed),with or without a history of smoking and alcohol consumption,hypertension,diabetes,heart disease,and trauma,duration of surgery(whether≤180 min),number of torn tendons(whether≥2),number of screws(whether≥2),large nodular osteophyte,and whether there was moderate-to-severe impingement.Single factor analysis was used to screen the factors influencing the need for opioid analgesics within 48 hours after arthroscopic rotator cuff repair.Results The results of univariate analysis showed that among the above independent variables,only the number of torn tendons≥2 was the factor affecting the need for opioid analgesics within 48 hours after arthroscopic rotator cuff repair(OR = 5.263,P = 0.007).Conclusions Patients with rotator cuff tears with≥2 tendons had more severe pain within 48 hours after rotator cuff repair,requiring opioid analgesics.Focus should be placed on postoperative pain in such patients,and a diverse analgesic regimen should be used early.

Innate immunity is the first line of the host cell defensing against viral infection,in which the pattern recognition re-ceptors(PRRs)such as Toll-like receptors(TLRs)and retinoic acid induces gene Ⅰ-like receptors(RLRs)play an important role.After virus infection,mitochondrial antiviral signaling protein(MAVS)in PRRs-mediated signaling pathway,which are of the com-mon linker molecule for downstream signal transmission,can receive signals transmitted by upstream TLR and RIG-Ⅰ,activate down-stream NF-κB and IRF3/7 signaling pathways leading to the activation of interferon(IFN)expression.Therefore MAVS acts as a bridge in the innate immune signaling pathway.More and more studies have shown that viruses have evolved a series of mechanism to escape the innate immune response over the long course of their evolution,and evaded the host's antiviral immune response by interfer-ing with multiple sites in the MAVS-mediated signaling pathway so as to complete its own replication and proliferation.In this paper,the role of MAVS in IFN-Ⅰ pathway and its latest research progress in the mechanism of anti-DNA viruses and anti-RNA viruses reac-tion are reviewed,providing theoretical basis for further studying the detailed mechanism of anti-virus of MAVS.

The classic formula Wuyaotang is the 49^th of the 100 formulas in the Catalogue of Ancient Classic Prescriptions (First Batch) issued by the National Administration of Traditional Chinese Medicine, and is from the Secrets from the Orchid Chamber (《兰室秘藏》) by LI Dongyuan of the Jin Dynasty. It is composed of Angelicae Sinensis Radix, Glycyrrhizae Radix et Rhizoma, Aucklandiae Radix, Linderae Radix, and Cyperi Rhizoma, and has the effect of moving Qi, regulating meridians, and relieving pain. It is mainly indicated for Qi stagnation and blood stasis syndrome. Based on the ancient books on Wuyaotang, this study systematically reviewed the formula source, composition, dosage, preparation, usage, functions, indications, preparation principle, drug processing, modification, etc. of Wuyaotang with the bibliometrics method, explored its historical evolution, and determined the key information. Statistical analysis of its modern literature shows that there are few studies of the original formula of Wuyaotang, and the clinical studies mainly focus on modified Wuyaotang. It has a wide range of treatment scope and can be used for the treatment of dysmenorrhea, delayed menstrual cycle, hypomenorrhea, and menstrual fever, as well as ulcerative colitis, spleen distortion, sciatica, child intestinal spasm, and other internal, surgical, gynecological, and pediatric diseases. The pathogenesis in traditional Chinese medicine (TCM) is Qi stagnation. Through the analysis and research on ancient books and modern literature recording Wuyaotang, this study is expected to provide a scientific basis for the clinical application, in-depth research, and development of the classic formula Wuyaotang.

Background Based on numerous epidemiological studies, radionuclide ¹³¹I can result in thyroid disease. Objective To study the contamination level of thyroid ¹³¹I in ¹³¹I treatment associated radiation workers in nuclear medicine departments in Shanghai. Methods Based on a general survey on basic situation of nuclear medicine in Shanghai, the level of internal exposure of radiation workers involved in ¹³¹I treatment was assessed with questionnaires and on-site monitoring. A portable γ spectrometer was used for on-site detection, the intake and annual effective dose were estimated according to the measurement results. Physical examination reports were collected for radionuclide positive workers, and chromosome testing was performed. Results There were 579 nuclear medicine workers and 175 were engaged in ¹³¹I treatment in Shanghai. ¹³¹I was detected in thyroid of 18 workers, with a detection rate of 10.3%. Among them, the detection rates of thyroid ¹³¹I in nurses and cleaners were 23% and 25% respectively. The mean measured activity of thyroid ¹³¹I was 306.1 Bq and the maximum measured activity was 3716.9 Bq (nurse). The maximum intake was estimated at 37544 Bq and the median was 786.4 Bq. The average annual committed effective dose was 1.22 mSv and the maximum value was 14.87mSv. The chromosomal aberration rates of all detected workers were negative, and all blood indicators were normal except one with low hemoglobin. Conclusion The annual dose of nuclear medical staff engaged in ¹³¹I treatment does not exceed the national standard limit, but the internal exposure of nurses and cleaners should not be ignored. On the basis of strengthening protection and decontamination, routine internal exposure monitoring should be carried out.

Background::Although overnight fasting is recommended prior to endoscopic retrograde cholangiopancreatography (ERCP), the benefits and safety of high-carbohydrate fluid diet (CFD) intake 2 h before ERCP remain unclear. This study aimed to analyze whether high-CFD intake 2 h before ERCP can be safe and accelerate patients’ recovery.Methods::This prospective, multicenter, randomized controlled trial involved 15 tertiary ERCP centers. A total of 1330 patients were randomized into CFD group ( n = 665) and fasting group ( n = 665). The CFD group received 400 mL of maltodextrin orally 2 h before ERCP, while the control group abstained from food/water overnight (>6 h) before ERCP. All ERCP procedures were performed using deep sedation with intravenous propofol. The investigators were blinded but not the patients. The primary outcomes included postoperative fatigue and abdominal pain score, and the secondary outcomes included complications and changes in metabolic indicators. The outcomes were analyzed according to a modified intention-to-treat principle. Results::The post-ERCP fatigue scores were significantly lower at 4 h (4.1 ± 2.6 vs. 4.8 ± 2.8, t = 4.23, P <0.001) and 20 h (2.4 ± 2.1 vs. 3.4 ± 2.4, t= 7.94, P <0.001) in the CFD group, with least-squares mean differences of 0.48 (95% confidence interval [CI]: 0.26–0.71, P <0.001) and 0.76 (95% CI: 0.57–0.95, P <0.001), respectively. The 4-h pain scores (2.1 ± 1.7 vs. 2.2 ± 1.7, t = 2.60, P = 0.009, with a least-squares mean difference of 0.21 [95% CI: 0.05–0.37]) and positive urine ketone levels (7.7% [39/509] vs. 15.4% [82/533], χ2 = 15.13, P <0.001) were lower in the CFD group. The CFD group had significantly less cholangitis (2.1% [13/634] vs. 4.0% [26/658], χ2 = 3.99, P = 0.046) but not pancreatitis (5.5% [35/634] vs. 6.5% [43/658], χ2 = 0.59, P = 0.444). Subgroup analysis revealed that CFD reduced the incidence of complications in patients with native papilla (odds ratio [OR]: 0.61, 95% CI: 0.39–0.95, P = 0.028) in the multivariable models. Conclusion::Ingesting 400 mL of CFD 2 h before ERCP is safe, with a reduction in post-ERCP fatigue, abdominal pain, and cholangitis during recovery.Trail Registration::ClinicalTrials.gov, No. NCT03075280.