Objective:To evaluate the alveolar ridge preservation effect of immediate implantation at extraction site with the im-proved CBCT measuring method.Methods:Eighty patients with extraction site were divided into test group A,B,C and control group.The patients were treated by means of immediate implant placement combined with large flap surgery,guided bone regener-ation(GBR)using mass Bio-Oss bone particles in the group A(the thickness of facial bone wall＜1 mm);The patients were trea-ted by means of immediate implant placement combined without flap surgery and bone graft in the group B(the thickness of facial bone wall ≥1 mm and＜2 mm)and C(the thickness of facial bone wall ≥2 mm),the CGF was implanted in the jumping space only when the thickness of jumping space was＞2 mm.In the control group,the alveolar sockets healed naturally without any in-tervention or treatment.CBCT was taken before surgery,immediately after surgery,and 6 months after surgery to evaluate the height and width of alveolar bone,the thickness of facial bone wall and jumping space.Results:The reduction of alveolar ridge height in group A,B,C and control group was(0.41±0.13,0.94±0.18,0.59±0.12,1.31±0.19)mm,The reduction of alveolar ridge width in group A,B,C and control group was(0.93±0.10,1.48±0.21,1.12±0.17,1.66±0.16)mum.The re-sults of four groups were statistically different(F=177.0,P＜0.001;F=125.3,P＜0.001).The alveolar ridge thickness of facial bone wall in group A,B,C and con-trol group was(0.98±0.25,2.39±0.28)mm,(1.43±0.52,2.10±0.33)mm,(2.17±0.41,2.79±0.27)mm before surgery and six months after immediate implantation.The results of each group were statistically different between before surgery and six months after immediate implantation(t=16.45,P＜0.001;t=7.357,P＜0.001;t=5.488,P＜0.001).Patients in three test groups had the thickness of jumping space＞2 mm and ≤2 mm,and the reduction of alveolar ridge width was(0.78±0.18,0.88±0.17)mm.The results were statistically different(t=17.18,P=0.018).Conclusion:The alveolar ridge preservation was obtained by means of immediate implant placement combined with large flap surgery,guided bone regeneration(GBR)using mass Bio-Oss bone particles at extraction site with the thickness of facial bone wall＜1 mm;The alveolar ridge preservation was obtained without flap surgery and GBR at extraction site with the thickness of facial bone wall≥1 mm.The preservation of soft and hard tissue was better in the axial palatal side of immediate implantation with the thickness of jumping space＞2 mm than that with the thickness of jumping space≤2 mm.

Objective To study the economic burden caused by healthcare-associated infection(HAI)in patients with severe acute pancreatitis(SAP),and provide theoretical basis for formulating HAI prevention and control measures.Methods Patients with SAP discharged from a tertiary first-class hospital in Jiangxi Province from July 1,2023 to June 30,2024 were selected as the study subjects.Information including demographic characteristics,clinical data,and hospitalization expense were collected.Patients were divided into a HAI group and a non-HAI group according to HAI occurrence.A propensity score matching(PSM)method was used to conduct a 1∶2 ma-tching,and differences in the length of hospital stay and hospitalization expense between the two groups of patients after PSM were compared.Results A total of 709 patients were included in the analysis,out of which 65 cases ex-perienced HAI,with a HAI incidence of 9.17％.After PSM,all 65 patients in the HAI group were successfully matched.The length of hospital stay,total hospitalization expense,expenses of medication and hygiene product of patients in the HAI group were all higher than those in the non-HAI group,and differences were all statistically sig-nificant(all P＜0.001).Patients who experienced≥2 episodes of HAI had a higher economic burden than those who experienced only once(P＜0.05).HAI of bloodstream,abdomen,digestive system,and respiratory system significantly increased the economic burden of patients(all P＜0.05).Conclusion HAI in SAP patients can extend the length of hospital stay and increase economic burden of patients.Targeted infection prevention and control mea-sures should be formulated to reduce the incidence of HAI and save medical resources.

Aim To investigate the effect and mechanism of angiotensin Ⅱ(Ang Ⅱ)on ferroptosis in white adi-pocytes.Methods The 3T3-L1 preadipocytes were differentiated into white adipocytes by inducer stimulation.The experiment was divided into control group,Ang Ⅱ group,Ang Ⅱ+Fer-1(ferroptosis inhibitor)group and Ang Ⅱ+PFT-α(p53 inhibitor)group.Ang Ⅱ was used to treat cells.RT-qPCR and Western blot were used to detect the expression levels of ferroptosis factors and adipokines.JC-1 kit was used to detect mitochondrial membrane potential(MMP)level.Iron ion kit was used to detect intracellular iron content.Glutathione(GSH)kit was used to detect GSH content.Fer-1 and Ang Ⅱ were added to treat cells to detect the the changes of ferroptosis level.The expression of p53 and spermidine/spermine N1-acetyltransferase 1(SAT1)protein was detected.Subsequently,PFT-α and Ang Ⅱ were added to co-treat cells to detect the changes of p53 and SAT1 protein expression,and to observe the effect of inhibiting p53 expression on the expression levels of ferroptosis factors and adipokines.Results 3T3-L1 cells were successfully differentiated into white adipocytes by stimulator-induced differentiation.Ang Ⅱ induced ferroptosis in white adipocytes.RT-qPCR results showed that compared with control group,the mRNA expression of anti-ferroptosis factor glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and iron regulatory protein 1(IRP-1)was down-regulated in Ang Ⅱ group,and the mRNA expression of pro-ferroptosis factor acyl-CoA synthetase of long-chain family member 4(ACSL4)was up-regulated.Western blot results showed that compared with control group,the protein expression of SLC7A11 and GPX4 was down-regulated in Ang Ⅱ group,and the protein expression of ACSL4 was up-regulated.Ang Ⅱ treatment increased the content of intracellular iron ions and decreased the levels of GSH and MMP.Compared with Ang Ⅱ group,the mRNA expression of IRP-1 and SLC7A11 was up-regulated in Ang Ⅱ+Fer-1 group.Ang Ⅱ induced changes in the expression profile of adipokines in white adipocytes.Western blot results showed that compared with control group,the protein ex-pression of pro-inflammatory adipokine leptin(LEP),resistin(RETN),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was up-regulated in Ang Ⅱ group,and the protein expression of anti-inflammatory adipokine adiponectin(AD-PN)and omentin 1(ITLN1)was down-regulated.In addition,Ang Ⅱ increased the protein expression of p53 and SAT1.Inhibition of p53 expression can improve the level of ferroptosis and adipokine expression in white adipocytes trea-ted with Ang Ⅱ.Western blot results showed that compared with Ang Ⅱ group,the protein expression of p53 and SAT1 was down-regulated in Ang Ⅱ+PFT-α group,the protein expression of SLC7A11 and GPX4 was up-regulated,and the protein expression of ACSL4 was down-regulated.The protein expression of ADPN was up-regulated in Ang Ⅱ+PFT-αgroup,and the protein expression of TNF-α,LEP and RETN was down-regulated.Conclusion Ang Ⅱ induces fer-roptosis in white adipocytes through activating the p53/SAT1 signaling pathway.

BACKGROUND: P16 inactivation is frequently accompanied by telomerase reverse transcriptase ( TERT ) amplification in human cancer genomes. P16 inactivation by DNA methylation often occurs automatically during immortalization of normal cells by TERT . However, direct evidence remains to be obtained to support the causal effect of epigenetic changes, such as P16 methylation, on cancer development. This study aimed to provide experimental evidence that P16 methylation directly drives cancer development. METHODS: A zinc finger protein-based P16 -specific DNA methyltransferase (P16-Dnmt) vector containing a "Tet-On" switch was used to induce extensive methylation of P16 CpG islands in normal human fibroblast CCD-18Co cells. Battery assays were used to evaluate cell immortalization and transformation throughout their lifespan. Cell subcloning and DNA barcoding were used to track the diversity of cell evolution. RESULTS: Leaking P16-Dnmt expression (without doxycycline-induction) could specifically inactivate P16 expression by DNA methylation. P16 methylation only promoted proliferation and prolonged lifespan but did not induce immortalization of CCD-18Co cells. Notably, cell immortalization, loss of contact inhibition, and anchorage-independent growth were always prevalent in P16-Dnmt&TERT cells, indicating cell transformation. In contrast, almost all TERT cells died in the replicative crisis. Only a few TERT cells recovered from the crisis, in which spontaneous P16 inactivation by DNA methylation occurred. Furthermore, the subclone formation capacity of P16-Dnmt&TERT cells was two-fold that of TERT cells. DNA barcoding analysis showed that the diversity of the P16-Dnmt&TERT cell population was much greater than that of the TERT cell population. CONCLUSION P16 methylation drives TERT -mediated immortalization and transformation of normal human cells that may contribute to cancer development.
Humans ; Telomerase/genetics* ; DNA Methylation/physiology* ; Fibroblasts/cytology* ; Cyclin-Dependent Kinase Inhibitor p16/metabolism* ; Cell Line ; Cell Transformation, Neoplastic/genetics*

Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

OBJECTIVE: To evaluate the effect and safety of Chinese medicine (CM) Fuzheng Huazhuo Decoction (FHD) in treating patients with coronavirus disease 2019 (COVID-19) who persistently tested positive for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). METHODS: This retrospective cohort study was conducted at Shanghai New International Expo Center shelter hospital in China between April 1 and May 30, 2022. Patients diagnosed as COVID-19 with persistently positive SARS-CoV-2 reverse transcription-polymerase chain reaction (RT-PCR) test results for ⩾8 days after diagnosis were enrolled. Patients in the control group received conventional Western medicine (WM) treatment, while those in the FHD group received conventional WM plus FHD for at least 3 days. The primary outcome was viral clearance time. Secondary outcomes included negative conversion rate within 14 days, length of hospital stay, cycle threshold (Ct) values of the open reading frame 1ab (ORF1ab) and nucleocapsid protein (N) genes, and incidence of new-onset symptoms during hospitalization. Adverse events (AEs) that occurred during the study period were recorded. RESULTS: A total of 1,765 eligible patients were enrolled in this study (546 in the FHD group and 1,219 in the control group). Compared with the control group, patients receiving FHD treatment showed shorter viral clearance time for nucleic acids [hazard ratio (HR): 1.500, 95% confidence interval (CI): 1.353-1.664, P<0.001] and hospital stays (HR: 1.371, 95% CI: 1.238-1.519, P<0.001), and a higher negative conversion rate within 14 days (96.2% vs. 82.6%, P<0.001). The incidence of new-onset symptoms was 59.5% in the FHD group, similar to 57.8% in the control group (P>0.05). The Ct values of ORF1ab and N genes increased more rapidly over time in the FHD group than those in the control group post-randomization (ORF1ab gene: β =0.436±0.053, P<0.001; N gene: β =0.415 ±0.053, P<0.001). The incidence of AEs in the FHD group was lower than that in the control group (24.2% vs. 35.4%, P<0.001). No serious AEs were observed. CONCLUSION FHD was effective and safe for patients with persistently positive SARS-CoV-2 PCR tests. (Registration No. ChiCTR2200063956).
Humans ; Drugs, Chinese Herbal/adverse effects* ; Retrospective Studies ; Male ; Female ; Middle Aged ; COVID-19 Drug Treatment ; SARS-CoV-2/drug effects* ; COVID-19/virology* ; Adult ; Aged ; Treatment Outcome

Aim To investigate the effect and mechanism of angiotensin Ⅱ(Ang Ⅱ)on ferroptosis in white adi-pocytes.Methods The 3T3-L1 preadipocytes were differentiated into white adipocytes by inducer stimulation.The experiment was divided into control group,Ang Ⅱ group,Ang Ⅱ+Fer-1(ferroptosis inhibitor)group and Ang Ⅱ+PFT-α(p53 inhibitor)group.Ang Ⅱ was used to treat cells.RT-qPCR and Western blot were used to detect the expression levels of ferroptosis factors and adipokines.JC-1 kit was used to detect mitochondrial membrane potential(MMP)level.Iron ion kit was used to detect intracellular iron content.Glutathione(GSH)kit was used to detect GSH content.Fer-1 and Ang Ⅱ were added to treat cells to detect the the changes of ferroptosis level.The expression of p53 and spermidine/spermine N1-acetyltransferase 1(SAT1)protein was detected.Subsequently,PFT-α and Ang Ⅱ were added to co-treat cells to detect the changes of p53 and SAT1 protein expression,and to observe the effect of inhibiting p53 expression on the expression levels of ferroptosis factors and adipokines.Results 3T3-L1 cells were successfully differentiated into white adipocytes by stimulator-induced differentiation.Ang Ⅱ induced ferroptosis in white adipocytes.RT-qPCR results showed that compared with control group,the mRNA expression of anti-ferroptosis factor glutathione peroxidase 4(GPX4),solute carrier family 7 member 11(SLC7A11)and iron regulatory protein 1(IRP-1)was down-regulated in Ang Ⅱ group,and the mRNA expression of pro-ferroptosis factor acyl-CoA synthetase of long-chain family member 4(ACSL4)was up-regulated.Western blot results showed that compared with control group,the protein expression of SLC7A11 and GPX4 was down-regulated in Ang Ⅱ group,and the protein expression of ACSL4 was up-regulated.Ang Ⅱ treatment increased the content of intracellular iron ions and decreased the levels of GSH and MMP.Compared with Ang Ⅱ group,the mRNA expression of IRP-1 and SLC7A11 was up-regulated in Ang Ⅱ+Fer-1 group.Ang Ⅱ induced changes in the expression profile of adipokines in white adipocytes.Western blot results showed that compared with control group,the protein ex-pression of pro-inflammatory adipokine leptin(LEP),resistin(RETN),interleukin-6(IL-6)and tumor necrosis factor-α(TNF-α)was up-regulated in Ang Ⅱ group,and the protein expression of anti-inflammatory adipokine adiponectin(AD-PN)and omentin 1(ITLN1)was down-regulated.In addition,Ang Ⅱ increased the protein expression of p53 and SAT1.Inhibition of p53 expression can improve the level of ferroptosis and adipokine expression in white adipocytes trea-ted with Ang Ⅱ.Western blot results showed that compared with Ang Ⅱ group,the protein expression of p53 and SAT1 was down-regulated in Ang Ⅱ+PFT-α group,the protein expression of SLC7A11 and GPX4 was up-regulated,and the protein expression of ACSL4 was down-regulated.The protein expression of ADPN was up-regulated in Ang Ⅱ+PFT-αgroup,and the protein expression of TNF-α,LEP and RETN was down-regulated.Conclusion Ang Ⅱ induces fer-roptosis in white adipocytes through activating the p53/SAT1 signaling pathway.

Objective:To evaluate the alveolar ridge preservation effect of immediate implantation at extraction site with the im-proved CBCT measuring method.Methods:Eighty patients with extraction site were divided into test group A,B,C and control group.The patients were treated by means of immediate implant placement combined with large flap surgery,guided bone regener-ation(GBR)using mass Bio-Oss bone particles in the group A(the thickness of facial bone wall＜1 mm);The patients were trea-ted by means of immediate implant placement combined without flap surgery and bone graft in the group B(the thickness of facial bone wall ≥1 mm and＜2 mm)and C(the thickness of facial bone wall ≥2 mm),the CGF was implanted in the jumping space only when the thickness of jumping space was＞2 mm.In the control group,the alveolar sockets healed naturally without any in-tervention or treatment.CBCT was taken before surgery,immediately after surgery,and 6 months after surgery to evaluate the height and width of alveolar bone,the thickness of facial bone wall and jumping space.Results:The reduction of alveolar ridge height in group A,B,C and control group was(0.41±0.13,0.94±0.18,0.59±0.12,1.31±0.19)mm,The reduction of alveolar ridge width in group A,B,C and control group was(0.93±0.10,1.48±0.21,1.12±0.17,1.66±0.16)mum.The re-sults of four groups were statistically different(F=177.0,P＜0.001;F=125.3,P＜0.001).The alveolar ridge thickness of facial bone wall in group A,B,C and con-trol group was(0.98±0.25,2.39±0.28)mm,(1.43±0.52,2.10±0.33)mm,(2.17±0.41,2.79±0.27)mm before surgery and six months after immediate implantation.The results of each group were statistically different between before surgery and six months after immediate implantation(t=16.45,P＜0.001;t=7.357,P＜0.001;t=5.488,P＜0.001).Patients in three test groups had the thickness of jumping space＞2 mm and ≤2 mm,and the reduction of alveolar ridge width was(0.78±0.18,0.88±0.17)mm.The results were statistically different(t=17.18,P=0.018).Conclusion:The alveolar ridge preservation was obtained by means of immediate implant placement combined with large flap surgery,guided bone regeneration(GBR)using mass Bio-Oss bone particles at extraction site with the thickness of facial bone wall＜1 mm;The alveolar ridge preservation was obtained without flap surgery and GBR at extraction site with the thickness of facial bone wall≥1 mm.The preservation of soft and hard tissue was better in the axial palatal side of immediate implantation with the thickness of jumping space＞2 mm than that with the thickness of jumping space≤2 mm.

Objective To study the economic burden caused by healthcare-associated infection(HAI)in patients with severe acute pancreatitis(SAP),and provide theoretical basis for formulating HAI prevention and control measures.Methods Patients with SAP discharged from a tertiary first-class hospital in Jiangxi Province from July 1,2023 to June 30,2024 were selected as the study subjects.Information including demographic characteristics,clinical data,and hospitalization expense were collected.Patients were divided into a HAI group and a non-HAI group according to HAI occurrence.A propensity score matching(PSM)method was used to conduct a 1∶2 ma-tching,and differences in the length of hospital stay and hospitalization expense between the two groups of patients after PSM were compared.Results A total of 709 patients were included in the analysis,out of which 65 cases ex-perienced HAI,with a HAI incidence of 9.17％.After PSM,all 65 patients in the HAI group were successfully matched.The length of hospital stay,total hospitalization expense,expenses of medication and hygiene product of patients in the HAI group were all higher than those in the non-HAI group,and differences were all statistically sig-nificant(all P＜0.001).Patients who experienced≥2 episodes of HAI had a higher economic burden than those who experienced only once(P＜0.05).HAI of bloodstream,abdomen,digestive system,and respiratory system significantly increased the economic burden of patients(all P＜0.05).Conclusion HAI in SAP patients can extend the length of hospital stay and increase economic burden of patients.Targeted infection prevention and control mea-sures should be formulated to reduce the incidence of HAI and save medical resources.