BACKGROUND:Copper is an essential trace element and plays a key role in series of physiological activities in the body.Metabolic disturbance of copper is closely associated with multiple diseases.Copper metabolism is mainly involved in the absorption,transport,storage and excretion of copper ions,and all the above processes regulate copper homeostasis in the body.In recent years,many studies have confirmed that copper homeostasis disorder severely affects the metabolic activities of the body and cause diseases in various systems.Besides,the role of copper in oral diseases has been of great interest.OBJECTIVE:To explore the role of copper in occurrence,development,and treatment of oral diseases,and provide a comprehensive overview of research advances in this field.METHODS:The first author searched relevant studies on copper in oral diseases using a computer in PubMed,Web of Science,and CNKI.The key words were"Cu,copper,copper metabolism,oral diseases,oral squamous cell carcinoma,periodontitis,oral submucous fibrosis,oral lichen planus,recurrent oral ulceration,pultitis"in English and Chinese.After screening,78 articles were included for further review and analysis.RESULTS AND CONCLUSION:(1)Patients with oral squamous cell carcinoma have elevated concentrations of copper in serum and saliva,and elevated copper promotes cancer progression through oxidative stress and promoting angiogenesis.Excessive elevation or reduction of copper concentration in tumor cells can inhibit the growth of tumor cells.The combination of copper and anticancer drugs can significantly improve the efficacy of drugs.(2)The concentration of copper in the serum of patients with periodontitis is increased,and excessive copper can aggravate periodontitis through promoting oxidative stress.Combination of copper and drugs can promote periodontal bone regeneration and periodontal tissue healing.(3)The level of copper is positively correlated with degree of oral mucosa fibrosis.Copper that enters the oral mucosa promotes fibrosis of oral mucosa by enhancing activity of lysyloxidase to increase production of collagen.(4)Copper levels are elevated in patients with oral lichen planus,and elevated copper may promote the progression of oral lichen planus by modulating immune cell function.(5)In patients with recurrent oral ulcers,serum copper level is significantly increased,and utilization of copper becomes disordered,which could decrease copper-containing enzyme activity,thus affecting the healing of ulcers.Copper is closely associated with multiple oral diseases and therapies targeting at copper could obviously enhance the therapeutic effect of drugs.But further studies are still needed to uncover its mechanisms to lay foundation for the better treatment of oral diseases.

BACKGROUND:Copper is an essential trace element and plays a key role in series of physiological activities in the body.Metabolic disturbance of copper is closely associated with multiple diseases.Copper metabolism is mainly involved in the absorption,transport,storage and excretion of copper ions,and all the above processes regulate copper homeostasis in the body.In recent years,many studies have confirmed that copper homeostasis disorder severely affects the metabolic activities of the body and cause diseases in various systems.Besides,the role of copper in oral diseases has been of great interest.OBJECTIVE:To explore the role of copper in occurrence,development,and treatment of oral diseases,and provide a comprehensive overview of research advances in this field.METHODS:The first author searched relevant studies on copper in oral diseases using a computer in PubMed,Web of Science,and CNKI.The key words were"Cu,copper,copper metabolism,oral diseases,oral squamous cell carcinoma,periodontitis,oral submucous fibrosis,oral lichen planus,recurrent oral ulceration,pultitis"in English and Chinese.After screening,78 articles were included for further review and analysis.RESULTS AND CONCLUSION:(1)Patients with oral squamous cell carcinoma have elevated concentrations of copper in serum and saliva,and elevated copper promotes cancer progression through oxidative stress and promoting angiogenesis.Excessive elevation or reduction of copper concentration in tumor cells can inhibit the growth of tumor cells.The combination of copper and anticancer drugs can significantly improve the efficacy of drugs.(2)The concentration of copper in the serum of patients with periodontitis is increased,and excessive copper can aggravate periodontitis through promoting oxidative stress.Combination of copper and drugs can promote periodontal bone regeneration and periodontal tissue healing.(3)The level of copper is positively correlated with degree of oral mucosa fibrosis.Copper that enters the oral mucosa promotes fibrosis of oral mucosa by enhancing activity of lysyloxidase to increase production of collagen.(4)Copper levels are elevated in patients with oral lichen planus,and elevated copper may promote the progression of oral lichen planus by modulating immune cell function.(5)In patients with recurrent oral ulcers,serum copper level is significantly increased,and utilization of copper becomes disordered,which could decrease copper-containing enzyme activity,thus affecting the healing of ulcers.Copper is closely associated with multiple oral diseases and therapies targeting at copper could obviously enhance the therapeutic effect of drugs.But further studies are still needed to uncover its mechanisms to lay foundation for the better treatment of oral diseases.

【Objective】 To analyze the results, characteristics and clinical value of video urodynamic study (VUD) of lower urinary tract symptoms (LUTS) in young male. 【Methods】 A total of 106 young male LUTS patients (18-45 years old) who received VUD in our hospital during Jan.2016 and Sep.2021 were collected to analyze the clinical and imaging urodynamic characteristics. 【Results】 Of the 106 patients, 55 (52.44%) had neurogenic lower urinary tract dysfunction (NLUTD)with clear neurological etiology, and 51 (48%) had non-neurogenic lower urinary tract dysfunction (NNLUTD). In NLUTD patients, dysuria was the most common symptom (76.74%); lumbosacral lesions were the main cause (76.36%); imaging urodynamics indicated weakening of detrusor muscle in different degrees. In NNLUTD patients,the main symptoms were frequent urination (48.72%) and dysuria (48.72%); about 58.97% of patients had two or more LUTS, and the main diagnosis was detrusor underactivity (DU)(35.90%). 【Conclusion】 NLUTD in young male is characterized by varying degrees of detrusor muscle weakness, detrusor sphincter dyscoordination, and decreased bladder compliance. NNLUTD is mostly caused by detrusor overactivity (DO) and DU.

Objective:To characterize the histopathological subtypes and their clinicopathological parameters of gender and onset age by common, rare and sparse primary esophageal malignant tumors (PEMT).Methods:A total of 272 437 patients with PEMT were enrolled in this study, and all of the patients were received radical surgery. The clinicopathological information of the patients was obtained from the database established by the State Key Laboratory of Esophageal Cancer Prevention & Treatment from September 1973 to December 2020, which included the clinical treatment, pathological diagnosis and follow-up information of esophagus and gastric cardia cancers. All patients were diagnosed and classified by the criteria of esophageal tumor histopathological diagnosis and classification (2019) of the World Health Organization (WHO). The esophageal tumors, which were not included in the WHO classification, were analyzed separately according to the postoperative pathological diagnosis. The χ 2 test was performed by the SPSS 25.0 software on count data, and the test standard α=0.05. Results:A total of 32 histopathological types were identified in the enrolled PEMT patients, of which 10 subtypes were not included in the WHO classification. According to the frequency, PEMT were divided into common (esophageal squamous cell carcinoma, ESCC, accounting for 97.1%), rare (esophageal adenocarcinoma, EAC, accounting for 2.3%) and sparse (mainly esophageal small cell carcinoma, malignant melanoma, etc., accounting for 0.6%). All the common, rare, and sparse types occurred predominantly in male patients, and the gender difference of rare type was most significant (EAC, male∶ female, 2.67∶1), followed with common type (ESCC, male∶ female, 1.78∶1) and sparse type (male∶ female, 1.71∶1). The common type (ESCC) mainly occurred in the middle thoracic segment (65.2%), while the rare type (EAC) mainly occurred in the lower thoracic segment (56.8%). Among the sparse type, malignant melanoma and malignant fibrous histiocytoma were both predominantly located in the lower thoracic segment (51.7%, 66.7%), and the others were mainly in the middle thoracic segment.Conclusion:ESCC is the most common type among the 32 histopathological types of PEMT, followed by EAC as the rare type, and esophageal small cell carcinoma and malignant melanoma as the major sparse type, and all of which are mainly occur in male patients. The common type of ESCC mainly occur in the middle thoracic segment, while the rare type of EAC mainly in the lower thoracic segment. The mainly sparse type of malignant melanoma and malignant fibrous histiocytoma predominately occur in the lower thoracic segment, and the remaining sparse types mainly occur in the middle thoracic segment.

Objective:To explore the related mechanisms of biological root resorption in decidual teeth by studying the biological effect of simulated occlusal force on the periodontal ligament stem cells (PDLSC) at different stages of root absorption.Methods:According to the tooth type and root absorption degree, healthy retained deciduous incisors and healthy first premolars that needed to be removed for orthodontic treatment were collected and divided into three groups with six teeth in each group: the deciduous unabsorbed group (UN group), the absorbed group (R group) and the permanent teeth group (P group). PDLSC was isolated from periodontal ligament and cultured. PDLSC of three groups were loaded with dynamic pressure of 0-45, 0-90, 0-135, 0-180, 0-225 and 0-270 kPa, respectively. The proliferation ability was detected by cell counting kit-8 (CCK-8) technique on day 1 to day 7, respectively. The apoptosis levels of PDLSC after loading with dynamic pressure of 0-45, 0-90, 0-135, 0-180 and 0-225 kPa were observed by the flow cytometry. The changes of microfilaments were observed by fibrous actin (F-actin) staining after the cytokeleton was subjected to dynamic pressure of 0-90 kPa.Results:PDLSC of three groups exhibited various proliferation abilities to dynamic pressure. The A values in the UN group and R group were significantly higher than those in the P group and the difference was statistically significant ( P<0.05). However, there was no significant difference between the UN group and the R group ( P>0.05). The A values of PDLSC in UN group and R group under dynamic pressures of 0-45, 0-90, 0-135 and 0-180 kPa had no statistical significance compared with the control group unloading dynamic pressure ( P>0.05). However, under 0-225 and 0-270 kPa dynamic pressures, the A values at the day 3 to day 7 were statistically significant ( P<0.05). The A values of PDLSC in P group under 0-45, 0-90, 0-135, 0-180 and 0-225 kPa dynamic pressures for 1 to 7 days were no statistically significant difference compared with the control group ( P>0.05). The A value in P group under the 0-270 kPa was statistically significant only on day 3 (1.386±0.131) and day 5 to day 7 (1.728±0.226, 2.029±0.168 and 2.263±0.210, respectively)( P<0.05). The result of apoptosis showed that the A values of PDLSC in UN group, R group and P group were significantly increased under 0-90, 0-135, 0-180 kPa and above dynamic pressures, respectively ( P<0.05) compared with the control group unloading dynamic pressure. Under 0-90 kPa dynamic pressure, F-actin fluorescence staining samples in three groups all showed green filaments which were arranged along the long axis of the cells in the R group and the P group, while some fibers in the UN group were closely arranged and promoted stress fiber assembly. Conclusions:The biological characteristics of PDLSC at different root absorption stages were changed when they were stimulated by mechanical stress, and PDLSC of the deciduous teeth at the root unabsorption stage were more sensitive to mechanical stress stimulation.

Osteosarcoma (OS) is the most common primary bone tumor in children and adolescents. It is an aggressive tumor with a tendency to spread to the lung, which is the most common site of metastasis. Patients with advanced OS with metastases have poor prognoses despite the application of chemotherapy, thus highlighting the need for novel therapeutic targets. The tumor microenvironment (TME) of OS is confirmed to be essential for and supportive of tumor growth and dissemination. The immune component of the OS microenvironment is mainly composed of tumor-associated macrophages (TAMs). In OS, TAMs promote tumor growth and angiogenesis and upregulate the cancer stem cell-like phenotype. However, TAMs inhibit the metastasis of OS. Therefore, much attention has been paid to investigating the mechanism of TAMs in OS development and the progression of immunotherapy for OS. In this article, we aim to summarize the roles of TAMs in OS and the major findings on the application of TAMs in OS treatment.

Objective:To construct an expression system of lentivirus vector encoding epidermal growth factor receptor-specific chimeric antigen receptor (EGFR-CAR) and programmed cell death ligand-1 (PD-L1) antibody.Methods:Human PD-L1-Fc protein was used to immunize BALB/c mice. Cell-fusion and subcloning were performed to screen stable hybridoma strains with high secretion of PD-L1-specific antibodies, which were identified by both ELISA and Western blot. The activity of the antibodies in blocking the binding of programmed cell death-1 (PD-1) to PD-L1 was determined by fluorescence-activated cell sorting (FACS). Antibody affinity was analyzed by Fortebio Octet96. A single-chain variable fragment (scFv) was further constructed after antibody full-length sequencing and humanization using CDR grafting method. Meanwhile, the genes encoding the light and heavy chain variable regions (VL and VH) were cloned from a hybridoma secreting antibody against human EGFR by 5′ RACE technology to construct scFv gene. The expression of scFv was confirmed using pcDNA3.1 vector. EGFR-CAR containing CD137 intracellular function domain and PD-L1-scFv was ligated using 2A gene. The synthetic single molecule was cloned into pLVX-EF1a-IRES-ZsGreen1 lentivirus expression vector, and then transfected into 293T cells using Lenti-X Packaging Single Shots (VSV-G) to prepare infectious virus. Expression of CAR on cell surface and the soluble form of PD-L1-scFv in the supernatant of transfected 293V cells were detected by FACS and ELISA.Results:A PD-L1 antibody named 11E3 with high ligand-receptor blocking performance was obtained. The humanized antibody showed a stable affinity (2.67×10 -10 mol/L) after directly grafting the mouse CDRs (CDR1, CDR2 and CDR3) to human frameworks. EGFR-scFv was effectively expressed in a form of Fc-fusion. Secretory CAR (CTZ0431-1) and membrane CAR (CTZ0431-2) expression plasmids were constructed using lentivirus vector containing EGFR-CAR and PD-L1-scFv. The infection efficiency in 293V cells was around 10%. EGFR-scFv on the cell membranes and PD-L1-scfv in the culture supernatants were detected after 293V cells were infected with CTZ0431-1. EGFR-scFv and PD-L1-scfv were expressed on the cell membranes of 293V cells infected with CTZ0431-2. The expression rate of CAR in LV-CART46407-1-transfected activated T cells was 39.3%. Conclusions:The lentivirus vectors co-expressing EGFR-CAR with moderate binding affinity and PD-L1-scFv with high binding affinity were successful constructed, which provided an essential tool for investing EGFR- and PD-L1 double targeted CAR-T cell therapy against solid tumor.

Objective To investigate the epidemiological characteristics and spatial distribution of hepatitis B in children in Kaizhou District of Chongqing from 2018 to 2019, and to provide a theoretical basis for the prevention and control of hepatitis B. Methods The prevalence of hepatitis B in Kaizhou District of Chongqing from 2018 to 2019 was calculated using Excel2003, and the spatial data was processed by SaTScan9.6. Aggregation of incidence of hepatitis B was investigated by calculating Moran^'s I coefficient. Aggregated distribution of the incidence of hepatitis B was explored by local spatial autocorrelation analysis. Results From 2018 to 2019, the cumulative number of hepatitis B in children in Kaizhou District of Chongqing was 86, with an average annual reported incidence rate of 3.01 per 100,000. Among 86 children with hepatitis B, 54 were male and 32 were female, with a male to female ratio of 1.69:1. The incidence rate of hepatitis B in 2019 was lower than that in 2018 ( P<0.05 ). The incidence rate of hepatitis B in males was higher than that in females in each year (P <0.05 ). The incidence rate of hepatitis B among children aged 10 to 14 years old was higher than that of any other age groups ( P<0.05 ). The hepatitis B incidence reported in Chongqing from 2018 to 2019 had spatial aggregation (Moran^'s I=0.4736, P<0.05; Moran^'s I=0.3915, P<0.05). The high-high (H-H) cluster areas of hepatitis B were mainly distributed in Maliu Township, Zishui Township, and Tianhe Town, while the low-low (L-L) cluster areas of hepatitis B were mainly distributed in Zhaojia Street and Changsha Town. Conclusion In 2018-2019, the incidence of hepatitis B among children in Kaizhou District, Chongqing City had a downward trend. The incidence of hepatitis B among males and children aged 10 to 14 years old was relatively high. There was a positive spatial autocorrelation of the incidence. The prevention and treatment of hepatitis B in high-risk population and hot spots should be strengthened.

Objective To compare a novel mirror-overlap method with the traditional manual measurement and the subjective assessment in assessing breast symmetry after oncoplastic surgery.Methods 59 patients with breast cancer who underwent conservation/reconstruction surgery were recruited.The post-operative assessment for breast symmetry was conducted using manual measurement,subjective assessment by the patient,their family member and a nurse,and the mirror-overlap method respectively.The latter method involved using the Photoshop to assess the area differences in breast outlines and shadows created by a flashlight when bilateral images were overlapped in the axis of midline.Results The distance differences between bilateral breasts measured using manual method were negatively correlated with symmetric coefficients in the mirror-overlap method (P＜0.05).There was a positive correlation between the average scores in the subjective assessment and symmetric coefficients of mirror-overlap method (P＜0.05).Conclusion The mirror-overlap method is an easy,feasible and cheap method which collects 3D breast information and provides an effective approach to assess breast symmetry after oncoplastic surgery.

Soft tissue sarcomas (STSs) are malignant tumors originating from mesenchymal tissues. They have lower morbidity rates than other cancers, which is only about 1% among adult malignancies. STSs consist of several subtypes and many of them are highly malignant. The 5-year survival rate is approximately 50%-60%. Investigation of STS pathogenesis demonstrated that epigenetic chang-es were closely related to the occurrence and development of STSs. Different from genetics changes, epigenetic changes are heritable and occur mainly through DNA methylation, histone modification, non-coding RNA regulation, and chromatin remodeling, all of which occur without changing the DNA sequence. Elucidation of the epigenetic mechanisms involved in STSs can help develop new clinical di-agnosis and treatment methods, and this article aims to review the recent advances in epigenetic research on STS.