Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objectives: Hypoparathyroidism (HP) is a rare endocrine disorder caused by parathyroid hormone (PTH) defi ciency. The PTH is a candidate gene for familial isolated hypoparathyroidism (FIH). This study aimed to investigate the pathogenicity of two novel rare variants (RVs) ofPTH through in vitro functional study. Methods: Targeted next-generation sequencing was used to identify candidate gene mutations. Clinical data were retrospectively collected. Wild-type (WT) PTH was used as a template for site-directed mutagenesis to create mutant eukaryotic expression plasmids, which were transfected into cells. Treated with or without 4-phenylbu tyric acid (4-PBA), the levels of intact PTH (iPTH) and PTH (1-84) were measured by chemiluminescence, and protein expression was assessed using Western blotting. Results: Two patients carrying PTH mutations (c.154G > A: p.Val52Ile, c.270G > T: p.Leu90Phe) were identified.Patient 1, a 45-year-old male, presented with carpal and pedal numbness, muscle cramps, and low serum calcium (1.29 mmol/L). Patient 2, a 12-year-old female, had muscle twitches, convulsions, low calcium (1.50 mmol/L), and iPTH of 4 pg/mL. The iPTH or PTH (1-84) levels in the medium transfected with mutant Val52Ile and Leu90Phe PTH decreased by 31%–38%, and 51%–96% compared to WT (allP < 0.05), which were not rescued by 4-PBA. No significant changes in intracellular PTH expression were observed. Conclusions In this study, two novel RVs of PTH(Val52Ile and Leu90Phe) were identified that may impair hormone synthesis and secretion. Our study has broadened the mutation spectrum of the PTH and shed light on potential mechanisms underlying FIH.

Objective:To investigate the expression and prognostic value of ANO1 in oral squamous cell carcinoma(OSCC)tissues.Methods:Immunohistochemistry(IHC,n=163)and Quantitative real-time PCR(qRT-PCR,n=42)were employed to detect the expression level of ANO1 protein and mRNA in OSCC tissues and paracancerous normal tissues.The relationship between ANO1 ex-pression and clinicopathological features(n=163)and prognosis(n=93)of the patients were analyzed,and the results were compared with those in TCGA database.Results:IHC and qRT-PCR confirmed that ANO1 was highly expressed in OSCC(P＜0.05),which was consistent with the results of the TCGA database.Cox regression analysis showed that ANO1 expression was significantly correlated with T stage,lymph node metastasis and TNM stage and poor prognosis(P＜0.05).By Cox regression analysis,ANO1 overexpression(P=0.002)and differentiation degree(P=0.034)were independent prognostic factors.Conclusion:ANO1 is highly expressed in OSCC and is correlated with poor prognosis,which can be used as a novel biomarker for poor prognosis of OSCC patients.

Oral diseases concern almost every individual and are a serious health risk to the popula-tion.The restorative treatment of tooth and jaw defects is an important means to achieve oral function and support the appearance of the contour.Based on the principle of"learning from the nature",Deng Xu-liang's group of Peking University School and Hospital of Stomatology has proposed a new concept of"microstructural biomimetic design and tissue adaptation of tooth/jaw materials"to address the worldwide problems of difficulty in treating dentine hypersensitivity,poor prognosis of restoration of tooth defects,and vertical bone augmentation of alveolar bone after tooth loss.The group has broken through the bottle-neck of multi-stage biomimetic technology from the design of microscopic features to the enhancement of macroscopic effects,and invented key technologies such as crystalline/amorphous multi-level assembly,ion-transportation blocking,and multi-physical properties of the micro-environment reconstruction,etc.The group also pioneered the cationic-hydrogel desensitizer,digital stump and core integrated restora-tions,and developed new crown and bridge restorative materials,gradient functionalisation guided tissue regeneration membrane,and electrically responsive alveolar bone augmentation restorative membranes,etc.These products have established new clinical strategies for tooth/jaw defect repair and achieved inno-vative results.In conclusion,the research results of our group have strongly supported the theoretical im-provement of stomatology,developed the technical system of oral hard tissue restoration,innovated the clinical treatment strategy,and led the progress of the stomatology industry.

Objective To explore the mechanism of Zhuangxuan Yin in the treatment of children with H1N1 pneumonia through regulating gut microbiota mediated by p38 mitogen-activated protein kinase(p38-MAPK)pathway.Methods A BALB/c mouse model of H1N1 pneumonia was prepared using the H1N1 influenza virus allantoic solution for nasal drop.The model was randomly separated into 5 groups:model group,Zhuangxuan Yin low-,medium-and high-dose groups,and high-dose Zhuangxuan Yin(28.66 g·kg-1)+anisomycin(10 mg·kg-1)group.The control group was infused with sterile physiological saline of equal volume using the same method.After treatment with Zhuangxuan Yin and anisomycin,the lung index of mice in each group was measured,and HE staining was applied to detect the pathological morphology of lung and large intestine tissues.16SrRNA gene sequencing was applied to detect the structural difference of gut microbiota in mice of each group.Enzyme-linked immunosorbent assay(ELASA)was applied to measure the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-4,IL-6,and IL-1β in bronchoalveolar lavage fluid(BALF)and large intestine tissue of mice in each group.Western Blot was applied to detect the expression of p38-MAPK pathway-related proteins in lung and large intestine tissues of mice in each group.Results Compared with the control group,the lung and large intestine tissues of the model group mice showed obvious pathological damage,the lung index,pathological score of lung and large intestine,ACE index,Shannon index,abundance of class Clostridia,the levels of TNF-α,IL-4,IL-6,and IL-1β in BALF and large intestine tissues,and p-p38-MAPK/p38-MAPK in lung and large intestine tissues increased(P＜0.05).The abundance of class Bacteroidales decreased(P＜0.05).Compared with the model group,the pathological damage in the lung and large intestine tissues of mice in the Zhuangxuan Yin low-,medium-and high-dose groups were reduced.The lung index,pathological score of lung and large intestine,ACE index,Shannon index,abundance of class Clostridia,the levels of TNF-α,IL-4,IL-6,and IL-1β in BALF and large intestine tissues,and p-p38-MAPK/p38-MAPK in lung and large intestine tissues decreased(P＜0.05),the abundance of class Bacteroidales increased(P＜0.05),and in a dose-dependent manner(P＜0.05).Compared with the high-dose Zhuangxuan Yin group,the pathological damage in the lung and large intestine tissues of mice in the high-dose Zhuangxuan Yin+anisomycin group was aggravated,the lung index,pathological score of lung and large intestine,ACE index,Shannon index,abundance of class Clostridia,the levels of TNF-α,IL-4,IL-6,and IL-1β in BALF and large intestine tissues,and p-p38-MAPK/p38-MAPK in lung and large intestine tissues increased(P＜0.05),and the abundance of class Bacteroidales decreased(P＜0.05).Conclusion Zhuangxuan Yin can improve the imbalance of intestinal microbiota in H1N1 pneumonia mice by inhibiting p38-MAPK signal activation,thereby inhibiting inflammation and reducing lung and large intestine tissue damage in mice,which may have a therapeutic effect on children with H1N1 pneumonia.

For the past few years, the indocyanine green fluorescence imaging has been widely used in the diagnosis and treatment of biliary tract diseases. Fluorescence visualization of the biliary system by indocyanine green accurately localize the diseased tissue and identify the biliary structures precisely, which effectively avoids the damage to the natural biliary structure and greatly improves the accuracy and safety of biliary surgery. However, the application of this new technology in biliary surgery is still at the exploratory stage, showing great potential for application while also exposing many problems and controversies. It is believed that with the continuous development and improvement, the indocyanine green fluorescence cholangiography will play a more important role in the diagnosis and treatment of biliary tract diseases in the future, and bring more benefits to patients.

Liquid chromatography-tandem mass spectrometry (LC-MS/MS) technology has the characteristics of high specificity and high throughput, making it rapidly applied and developed in the field of clinical testing. Its application in the monitoring of therapeutic drugs can effectively improve the quantitative accuracy and sensitivity, and formulate a personalized and optimal dosing plan for patients. However, this technology still faces some challenges, and automation, quality control, and quantitative traceability will be the future development direction.