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.

Patients with primary hyperparathyroidism (PHPT) have an increased risk of kidney stones. Parathyroidectomy (PTX) is the main treatment for PHPT. However, the effect of PTX on kidney stones in patients with PHPT is unclear. This article summarizes the risk factors for kidney stones after PTX or not and the influence of PTX on the occurrence and development of renal stones in patients with PHPT, in order to explore the causes and mechanisms of PHPT and its postoperative renal stones.

Patients with primary hyperparathyroidism (PHPT) have an increased risk of kidney stones. Parathyroidectomy (PTX) is the main treatment for PHPT. However, the effect of PTX on kidney stones in patients with PHPT is unclear. This article summarizes the risk factors for kidney stones after PTX or not and the influence of PTX on the occurrence and development of renal stones in patients with PHPT, in order to explore the causes and mechanisms of PHPT and its postoperative renal stones.

AIM:To investigate the effects of emo-din on autophagy and apoptosis in rats with severe pneumonia(KP)caused by K.pneumoniae and its possible mechanism.METHODS:The KP rat model was established by infecting K pneumonia was treat-ed with Emodin.The rats were grouped into Sham surgery group,KP group,low concentration Emodin group,medium concentration Emodin group,high concentration Emodin group,and Emodin+sirtinol(SIRT1 activity inhibitor)group;Arterial partial pres-sure of carbon dioxide(PaCO2),arterial partial pres-sure of oxygen(PaO2)and arterial oxygen saturation(SaO2)were measured by blood gas analyzer;the white blood cells and neutrophils in bronchoalveo-lar lavage fluid(BALF)were measured by Wright-Gi-emsa staining;HE staining was applied to detect pathological changes in lung tissue in each group;ELISA was applied to detect the expression of IL-6,TNF-α,and IL-1β in lung tissues of each group;elec-tron microscopy scanning was applied to observe the autophagy of cells in lung tissues of each group;the expression of LC3B in lung tissues was observed by immunofluorescence staining;TUNEL method was applied to detect changes in cell apoptosis in lung tissue of rats in each group;Western blot was applied to detect the expression of silent informa-tion regulatory factor(SIRT1),adenosine monophos-phate activated protein kinase(AMPK),LC3-Ⅱ,LC3-Ⅰ,c-caspase-3,and caspase-3 proteins in lung tissue.RESULTS:K.pneumoniae caused severe lung tissue damage in rats with pneumonia,increased inflam-matory infiltration and cytokine release in the lungs,arterial blood PaO2 and SaO2 levels de-creased,PaCO2 levels increased,white blood cells and neutrophils count increased in BALF,increased cell apoptosis rate and c-caspase-3/caspase-3 level,and the cell autophagy and the levels of autophagy related proteins LC3-Ⅱ/LC3-Ⅰ were decreased(all P＜0.05),after Emodin treatment,SIRT1/AMPK signal-ing pathway was activated,PaO2 and SaO2 levels in arterial blood were increased,PaCO2 levels was de-creased,inflammatory reaction was inhibited,cell apoptosis in lung tissue was inhibited(all P＜0.05),and cell autophagy level was restored,sirtinol,a SIRT1 inhibitor,partially reversed the therapeutic ef-fect of Emodin on KP rats after inhibiting SIRT1/AMPK signaling pathway(P＜0.05).CONCLUSION:Emodin may enhance autophagy of lung tissue cells and inhibit apoptosis of rat lung tissue cells by acti-vating SIRT1/AMPK pathway,which may provide po-tential therapeutic options for KP.

Objective:To investigate the effects and potential mechanisms of melatonin on cognitive dysfunction induced by long-term anesthesia with isoflurane.Methods:Male C57BL/6J mice aged 2 months were divided into control group, isoflurane group, melatonin group, and isoflurane+ melatonin group by random number table, with 6 mice in each group.Three days after anesthesia, cognitive function of mice was assessed by Y-maze and fear conditioning (FC) tests. ATP content in the hippocampus was measured by an ATP assay kit. Western blot was used to detect the expression of DRP1, pDRP1, MFN2, pAMPK and SIRT1 proteins in the hippocampus. Cultured HT-22 cells derived from mouse hippocampal neurons in vitro were divided into control group, isoflurane group, melatonin group, and isoflurane + melatonin group, and the levels of reactive oxygen species (ROS) in each group were detected by flow cytometry after intervention. Statistical analysis was performed by SPSS 22.0 software, and one-way ANOVA was used for comparisons among multiple groups.Results:(1) There was a statistically significant difference in the percentage of freezing behavior in contextual fear memory among the four groups of mice ( F=39.09, P<0.05). The percentage of freezing behavior in the isoflurane group was lower than that in the control group ((44.23±8.88)% vs (75.87±5.90)%, P<0.05), while the percentage of freezing behavior in the isoflurane+ melatonin group((67.45±14.89)%)was higher than that in the isoflurane group ( P<0.05). There was also a statistically significant difference in the percentage of exploration in the novel arm among the four groups of mice ( F=13.87, P<0.05). The percentage of exploration in the novel arm in the isoflurane group was lower than that in the control group((33.64±6.53)% vs (47.13±3.87)%, P<0.05), while the percentage of exploration in the novel arm in the isoflurane+ melatonin group((43.05±1.64)%)was higher than that in the isoflurane group ( P<0.05). (2) Statistically significant difference in the levels of ATP in the hippocampus was found among the four groups of mice ( F=49.22, P<0.05). The level of ATP in the hippocampus in the isoflurane group was lower than that in the control group((2.29±0.15)nmol/mg vs (3.58±0.12)nmol/mg, P<0.05), while the level of ATP in the hippocampus in the isoflurane+ melatonin group ((3.02±0.27)nmol/mg)was higher than that in the isoflurane group ( P<0.05). There was a statistically significant difference in the levels of ROS in HT-22 cells among the four groups ( F=18.36, P<0.05). The level of ROS in HT-22 cells in the isoflurane group was higher than that in the control group after anesthesia ( P<0.05), while the level of ROS in HT-22 cells in the isoflurane+ melatonin group was lower than that in the isoflurane group after anesthesia ( P<0.05). (3) There were statistically significant difference in the levels of pDRP1, pAMPK and SIRT1 protein in the hippocampus among the four groups of mice ( F=19.87, 21.20, 25.65, all P<0.05). The levels of pDRP1 and SIRT1 protein in the hippocampus in the isoflurane group were both lower than those in the control group (both P<0.05), while the levels of pDRP1 and SIRT1 protein in the hippocampus in the isoflurane+ melatonin group were both higher than those in the isoflurane group (both P<0.05). In the isoflurane group, the expression of pAMPK protein in the hippocampal region was higher than that in the control group ( P<0.05), while the expression of pAMPK protein in the isoflurane+ melatonin group was lower than that in the isoflurane group ( P<0.05). Conclusion:Melatonin improves long-term isoflurane anesthesia-induced cognitive dysfunction by regulating mitochondrial homeostasis through the AMPK/SIRT1 signaling pathway.

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.

AIM:To investigate the effects of emo-din on autophagy and apoptosis in rats with severe pneumonia(KP)caused by K.pneumoniae and its possible mechanism.METHODS:The KP rat model was established by infecting K pneumonia was treat-ed with Emodin.The rats were grouped into Sham surgery group,KP group,low concentration Emodin group,medium concentration Emodin group,high concentration Emodin group,and Emodin+sirtinol(SIRT1 activity inhibitor)group;Arterial partial pres-sure of carbon dioxide(PaCO2),arterial partial pres-sure of oxygen(PaO2)and arterial oxygen saturation(SaO2)were measured by blood gas analyzer;the white blood cells and neutrophils in bronchoalveo-lar lavage fluid(BALF)were measured by Wright-Gi-emsa staining;HE staining was applied to detect pathological changes in lung tissue in each group;ELISA was applied to detect the expression of IL-6,TNF-α,and IL-1β in lung tissues of each group;elec-tron microscopy scanning was applied to observe the autophagy of cells in lung tissues of each group;the expression of LC3B in lung tissues was observed by immunofluorescence staining;TUNEL method was applied to detect changes in cell apoptosis in lung tissue of rats in each group;Western blot was applied to detect the expression of silent informa-tion regulatory factor(SIRT1),adenosine monophos-phate activated protein kinase(AMPK),LC3-Ⅱ,LC3-Ⅰ,c-caspase-3,and caspase-3 proteins in lung tissue.RESULTS:K.pneumoniae caused severe lung tissue damage in rats with pneumonia,increased inflam-matory infiltration and cytokine release in the lungs,arterial blood PaO2 and SaO2 levels de-creased,PaCO2 levels increased,white blood cells and neutrophils count increased in BALF,increased cell apoptosis rate and c-caspase-3/caspase-3 level,and the cell autophagy and the levels of autophagy related proteins LC3-Ⅱ/LC3-Ⅰ were decreased(all P＜0.05),after Emodin treatment,SIRT1/AMPK signal-ing pathway was activated,PaO2 and SaO2 levels in arterial blood were increased,PaCO2 levels was de-creased,inflammatory reaction was inhibited,cell apoptosis in lung tissue was inhibited(all P＜0.05),and cell autophagy level was restored,sirtinol,a SIRT1 inhibitor,partially reversed the therapeutic ef-fect of Emodin on KP rats after inhibiting SIRT1/AMPK signaling pathway(P＜0.05).CONCLUSION:Emodin may enhance autophagy of lung tissue cells and inhibit apoptosis of rat lung tissue cells by acti-vating SIRT1/AMPK pathway,which may provide po-tential therapeutic options for KP.

Objective:To report the nationwide surveillance results of pathogenic profiles and antimicrobial resistance patterns of Gram-positive bloodstream infections in China in 2023.Methods:The clinical isolates of Gram-posttive bacteria from blood cultures were collected in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System（BRICS）during January to December 2023. Antimicrobial susceptibility testing was performed using the dilution method recommended by the Clinical and Laboratory Standards Institute（CLSI）. Statistical analyses were conducted using WHONET 5.6 and SPSS 25.0 software.Results:A total of 4 385 Gram-positive bacterial isolates were obtained from 60 participating center. The top five pathogens were Staphylococcus aureus（ n=1 544，35.2%），coagulase-negative Staphylococci（ n=1 441，32.9%）， Enterococcus faecium（ n=574，13.1%）， Enterococcus faecalis（ n=385，8.8%），and α-hemolytic Streptococci（ n=187，4.3%）. The prevalence of methicillin-resistant Staphylococcus aureus（MRSA）and methicillin-resistant coagulase-negative Staphylococci（MRCNS）was 26.2%（405/1 544）and 69.8%（1 006/1 441），respectively. Notably，all Staphylococci remained susceptible to glycopeptide or daptomycin. Staphylococcus aureus demonstrated excellent susceptibility（>97.0%）to cephalobiol，rifampicin，trimethoprim-sulfamethoxazole，linezolid，minocycline，tigecycline，and eravacycline. No Enterococcus exhibiting resistance to linezolid were detected. Glycopeptide resistance was uncommon but more frequent in Enterococcus faecium（resistance to vancomycin and teicoplanin：both 1.7%）compared to Enterococcus faecalis（both 0.3%）. The detection rates of MRSA and MRCNS exhibited significant regional variations across the country（ χ2=17.674 and 148.650，respectively，both P<0.001）. No vancomycin-resistant Enterococci were detected in central China. Institutional comparison demonstrated higher prevalence of MRSA（ χ2=14.111， P<0.001）and MRCNS（ χ2=4.828， P=0.028）in provincial hospitals than that in municipal hospitals. Socioeconomic analysis identified elevated detection rates of both MRSA（ χ2=18.986， P<0.001）and MRCNS（ χ2=4.477， P=0.034）in less developed regions（per capita GDP

Objective:To report the results of bacterial resistant investigation collaborative system（BRICS）on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2023，and provide reference for clinical tretment of bloodstream infections and prevention and control of bacterial resistance.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of BRICS were collected during January 2023 to December 2023. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 were used to analyze the data.Results:During the study period，11 492 strains of Gram-negative bacteria were collected from 60 hospitals，of which 10 098（87.9%）were Enterobacterales and 1 394（12.1%）were non-fermentative bacteria. The top 5 bacterial species were Escherichia coli（50.0%）， Klebsiella pneumoniae（26.1%）， Pseudomonas aeruginosa（5.1%）， Acinetobacter baumannii complex（5.0%）and Enterobacter cloacae complex（4.1%）. The ESBL-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus mirablilis were 46.8%（2 685/5 741），18.3%（549/2 999）and 44.0%（77/175），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（76/5 741）and 15.0%（450/2 999）；32.9%（25/76）and 78.0%（351/450）of CREC and CRKP were sensitive to ceftazidime/avibactam combination，respectively. 94.7%（72/76）and 90.2%（406/450）of CREC and CRKP were sensitive to aztreonam/avibactam combination. Furthermore，57.9%（44/76）and 79.1%（356/450）were sensitive to imipenem/relebactam combination. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 64.6%（370/573），while more than 80.0% of CRAB complex was sensitive to tigecycline，eravacycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 17.0%（99/581）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of important Gram-negative bacteria resistance among different regions in China，with statistically significant differences in the prevalence of CREC，CRKP，CRPA and CRAB complex（ χ2=10.6，28.6，10.8 and 19.3， P<0.05）. The prevalence of ESBL-producing Escherichia coli， CREC，CRAB complex and CRKP were higher in provincial hospitals than those in municipal hospitals（ χ2=12.5，9.8，12.7 and 57.8，all P<0.01）. Conclusions:Gram-negative bacteria are the main pathogens causing bloodstream infections in China，and Escherichia coli is ranked in the top，while the trend of Klebsiella pneumoniae increases continuously with time. CRKP infection shows a slow upward trend，CREC infecton maintains a low prevalence level，and CRAB complex infection continues to exhibit a high prevalence rate. The composition and resistance patterns of pathogens causing bloodstream infections vary to some extent across different regions and levels of hospitals in China.

Objective:To evaluate the effectiveness and safety of zoledronic acid in patients with Paget′s disease of bone based on clinical data from a single medical center.Methods:This retrospective study included 24 patients diagnosed with Paget′s disease of bone and treated with zoledronic acid at Peking Union Medical College Hospital between January 2009 and June 2020. Demographic data, clinical symptoms, treatment efficacy, and safety outcomes were collected. The primary efficacy measure was serum alkaline phosphatase(ALP) levels. Treatment was considered effective if ALP levels returned to normal or decreased by more than 75% from baseline in the difference between the ALP level and its normal median value.Results:Among the 24 patients with Paget′s bone disease, the most commonly affected site was the skull(in 17 cases). All patients received a single 5 mg intravenous infusion of zoledronic acid. Serum ALP levels significantly decreased after treatment. Among the 15 patients who completed at least 3 months of follow-up, all achieved treatment success. The median time for serum ALP levels to reach the target was 13.1(9.4, 26.1) weeks. In 12 patients, ALP levels normalized within a medium of 16.9(11.5, 37.3) weeks, and remained stable over a medium follow-up of 4.56(2.42, 5.71) years. The most common side effects were hypocalcemia(21 cases, 87.5%) and flu-like symptoms(17 cases, 70.8%). Seven patients had severe hypocalcemia(serum calcium<1.75 mmol/L), and they had higher baseline levels of ALP, calcium, and phosphorus compared to those with mild hypocalcemia.Conclusions:Zoledronic acid 5 mg intravenous infusion effectively controlled disease activity in patients with Paget′s disease of bone. Generally, Most patients achieved treatment goals within 3-4 months, with sustained remission for a median of 4 years. Hypocalcemia was the most frequent side effect, underscoring the importance of timely calcium and vitamin D supplementation.