BACKGROUND:Carnosic acid,a bioactive compound found in rosemary,has been shown to reduce inflammation and reactive oxygen species(ROS).However,its mechanism of action in osteoclast differentiation remains unclear. OBJECTIVE:To investigate the effects of carnosic acid on osteoclast activation,ROS production,and mitochondrial function. METHODS:Primary bone marrow-derived macrophages from mice were extracted and cultured in vitro.Different concentrations of carnosic acid(0,10,15,20,25 and 30 μmol/L)were tested for their effects on bone marrow-derived macrophage proliferation and toxicity using the cell counting kit-8 cell viability assay to determine a safe concentration.Bone marrow-derived macrophages were cultured in graded concentrations and induced by receptor activator of nuclear factor-κB ligand for osteoclast differentiation for 5-7 days.The effects of carnosic acid on osteoclast differentiation and function were then observed through tartrate-resistant acid phosphatase staining,F-actin staining,H2DCFDA probe and mitochondrial ROS,and Mito-Tracker fluorescence detection.Western blot and RT-PCR assays were subsequently conducted to examine the effects of carnosic acid on the upstream and downstream proteins of the receptor activator of nuclear factor-κB ligand-induced MAPK signaling pathway. RESULTS AND CONCLUSION:Tartrate-resistant acid phosphatase staining and F-actin staining showed that carnosic acid dose-dependently inhibited in vitro osteoclast differentiation and actin ring formation in the cell cytoskeleton,with the highest inhibitory effect observed in the high concentration group(30 μmol/L).Carnosic acid exhibited the most significant inhibitory effect during the early stages(days 1-3)of osteoclast differentiation compared to other intervention periods.Fluorescence imaging using the H2DCFDA probe,mitochondrial ROS,and Mito-Tracker demonstrated that carnosic acid inhibited cellular and mitochondrial ROS production while reducing mitochondrial membrane potential,thereby influencing mitochondrial function.The results of western blot and RT-PCR revealed that carnosic acid could suppress the expression of NFATc1,CTSK,MMP9,and C-fos proteins associated with osteoclast differentiation,and downregulate the expression of NFATc1,Atp6vod2,ACP5,CTSK,and C-fos genes related to osteoclast differentiation.Furthermore,carnosic acid enhanced the expression of antioxidant enzyme proteins and reduced the generation of ROS during the process of osteoclast differentiation.Overall,carnosic acid exerts its inhibitory effects on osteoclast differentiation by inhibiting the phosphorylation modification of the P38/ERK/JNK protein and activating the MAPK signaling pathway in bone marrow-derived macrophages.

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

AIM:The objective of this study is to examine the expression of profilin 1(PFN1)in mice with di-abetic nephropathy and determine its association with immune cell infiltration.METHODS:This study presents an analy-sis of PFN1 expression and immune cell infiltration in patients with diabetic nephropathy,utilizing transcriptome expres-sion data from kidney tissue microarray.Additionally,the findings were validated in a diabetic nephropathy mouse model.Sixteen C57BL/6 mice were randomly assigned into two groups,namely the normal group and the model group,in an equal manner.The model group underwent the establishment of the diabetic nephropathy model through intraperitoneal injection of streptozotocin.Subsequently,the expression levels of CD11b,F4/80,CC chemokine receptor 4(CCR4),interleukin-1 receptor type I(IL-1R1),B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-3 in kidney tissue were assessed upon successful establishment of the diabetic nephropathy model.Furthermore,the overexpression of PFN1 was observed in a cellular model of diabetic nephropathy,and the protein expression levels of monocyte chemotactic pro-tein-1(MCP-1)and caspase-3 were assessed.RESULTS:The expression of PFN1 was found to be significantly in-creased in the GSE30122 dataset of transcriptome expression in kidney tissues affected by diabetic nephropathy(P＜0.01).This increase in PFN1 expression was found to be correlated with the presence of macrophages and T cells.Fur-thermore,the renal tissue of the diabetic nephropathy model group exhibited significant pathological changes.In this mod-el group,the expression levels of PFN1,CD11b,F4/80,CCR4,IL-1R1,Bax,Bcl-2,and caspase-3 were all significant-ly increased(P＜0.01).Overexpression of PFN1 could enhance the expression of MCP-1 and caspase-3 proteins.CON-CLUSION:Macrophages and Th17 cells were identified within the renal tissue of mice with diabetic nephropathy,con-comitant with an up-regulation in the expression of PFN1.This up-regulation was observed to facilitate the induction of apoptosis in the context of diabetic nephropathy.

Humans ; Retrospective Studies ; Tetanus/epidemiology* ; China/epidemiology*

Objective:To evaluate our self-designed pubic symphysis orthotic compression anatomic plate (PSOCAP) in the treatment of fractures and dislocations around the pubic symphysis.Methods:A retrospective study was conducted to analyze the 16 patients with fracture or dislocation around the pubic symphysis who had been treated by our self-designed PSOCAP at Department of Traumatic Surgery, Center for Orthopaedic Surgery, The Third Hospital Affiliated to Southern Medical University from January 2021 to June 2022. There were 8 males and 8 females with an age of (41±19) years. According to the Tile classification for pelvic fractures, there were 3 cases of type B1, 2 cases of type B3, 1 case of type C1.2, 4 cases of type C1.3, 3 cases of type C2, and 3 cases of type C3. There were 8 cases of pubic symphysis separation and 8 fractures of the pubic ramus (2 ones at Nakatani zone Ⅰ and 6 ones at Nakatani zone Ⅱ). Time from injury to surgery was 16 (11, 53) days, ranging from 4 to 348 days. The fractures or dislocations around the pubic symphysis were exposed by the modified Stoppa approach, reduced with the assistance of PSOCAP and fixated with PSOCAP; the posterior pelvic ring was reduced and fixated by corresponding surgical methods. Recorded were the surgical time, intraoperative bleeding, postoperative quality of fracture reduction, surgical complications, and functional recovery at the last follow-up concerning the pelvic anterior ring.Results:Surgery went on successfully in the 16 patients. Their surgical time was (58±15) min, ranging from 40 to 90 min, and their intraoperative bleeding 85 (63, 150) mL, ranging from 50 to 250 mL. According to the Matta scoring, the fracture reduction was evaluated as excellent in 10 cases, as good in 3 cases and as fair in 3 cases. The (10±3)-month follow-up for the 16 patients revealed complete fracture union for all after (12±2) weeks. According to the Majeed scoring at the last follow-up, the pelvic function was evaluated as excellent in 5 cases, as good in 7 cases, and as fair in 4 cases. No such postoperative complications as fracture displacement or internal fixation failure occurred.Conclusion:Owing to the biplane and integrated structure, our self-designed PSOCAP can help reduce the fractures or dislocations around the pubic symphysis to achieve anatomical reduction and strong internal fixation, leading to good clinical efficacy.

Abstract: Objective　To investigate the impacts of vaccination with inactivated SARS-COV-2 vaccine on the clinical manifestations and serological responses of COVID-19 patients infected by Delta and Alpha variants. Methods　Clinical and experimental data of 341 confirmed SARS-COV-2 patients were collected from The Eighth Affiliated Hospital of Guangzhou Medical University May 1- September 30, 2021. The subjects were divided into Delta and Alpha variant group according to virus variants, and were divided into vaccinated group and unvaccinated group according to whether they had received inactivated COVID-19 vaccine or not. The clinical manifestations and serological responses of patients with Delta and Alpha variant, and vaccinated and unvaccinated patients with Delta and Alpha variants were compared. Results　Totally 253 patients were infected with Delta variant (103 vaccinated and 150 unvaccinated patients), and 88 patients were infected with Alpha variant (21 vaccinated and 67 unvaccinated patients). The proportion of asymptomatic infection in Delta variants group was significantly lower than that in Alpha variants group (P<0.01). Delta variant group of vaccination rates and vaccine breakthrough infection rate was 40.7% (103/253) and 22.9% (58/253), were higher than Alpha variant group was 23.9% (21/88) and 8.0% (7/88), difference was statistically significant (χ2= 8.009, 9.484, P<0.01). The proportion of cough and fever in Delta variant group was higher than that in Alpha variant group (both P<0.01), the peak viral load was higher than that of Alpha variant group (P<0.01), the virus duration was longer than that of Alpha variant group (P<0.01), the levels of SAA, CRP and IFN were higher than those of Alpha variant group (all P<0.05), CD4+T cell count was lower than that of Alpha variant group (P<0.05), IgG and IgM levels were lower than those of Alpha variant group (both P<0.01). The proportion of moderate COVID-19 in the vaccinated group was lower than that in the unvaccinated group (P<0.01). In these two variants, the peak viral load of vaccinated group was lower than that of the unvaccinated group (both P<0.01), the duration of virus was shorter than that of unvaccinated group (both P<0.01). The levels of SAA, CRP and IL-6 in the vaccinated group were lower than those in the unvaccinated group (all P<0.05), CD4+T cell level was higher than that of unvaccinated group (both P<0.05), IgG and IgM level were higher than those in unvaccinated group (both P<0.05). Conclusions　Delta variant can lead to higher viral load and more severe disease course, which is associated with vaccine breakthrough infection. Inactivated vaccines for COVID-19 can reduce severe illness and death by reducing viral load, disease duration and inflammatory response through humoral and cellular immune mechanisms.

OBJECTIVE: To analyze the risk factors affecting prognosis of children with hemophagocytic lymphohistiocytosis (HLH). METHODS: The clinical manifestations and laboratory data of 143 HLH children who met the HLH-2004 diagnostic criteria in Shenzhen Children's Hospital from January 2009 to May 2017 were retrospectively analyzed, and the independent factors affecting prognosis were also analyzed. RESULTS: The median age of 143 HLH children was 1.9 (0.1-14.3) years old, and the median follow-up time was 6.7 years (1 day - 11.9 years). The overall survival rate of 1 month, 1 year, and 10 years was (87.4±5.5)%, (81.1±6.5)%, and (81.1±6.5)%, respectively. The deaths occurred within 1 year after onset. Multivariate analysis showed that central nervous system (CNS) involvement (P=0.047), low hemoglobin (P=0.002), prolonged activated partial thromboplastin time (APTT) (P<0.001), high triglyceride (P=0.005) were all the independent risk factors affecting survival of the children. Receiver operating characteristic curve indicated that APTT (AUC=0.753, P<0.001) was more valuable than other risk factors in predicting death of the children. The cut-off value of APTT was 56.6 s, and the sensitivity and specificity of which was 55.6% and 89.7%, respectively. CONCLUSION Hypohemoglobinemia, prolonged APTT, hypertriglyceridemia, and CNS involvement the risk factors affecting prognosis of HLH, and prolonged APTT shows a strong predictive value for death.
Adolescent ; Child ; Child, Preschool ; Humans ; Infant ; Lymphohistiocytosis, Hemophagocytic ; Prognosis ; Retrospective Studies ; Risk Factors ; Survival Rate

Betacoronavirus ; Coronavirus Infections ; Humans ; Pandemics ; Pneumonia, Viral ; Single-Cell Analysis

Objective To investigate the epidemiological characteristics and clinic symptoms of an outbreak of infectious diarrhea caused by Sapovirus,in CT community,Zhaoqing,Guangdong province.Methods Retrospective study of field epidemiology investigation was carried out and Chi-square test was used enteroviruses were detected by RT-PCR in swab,fecal specimens and water samples.Results A total of 422 cases were identified in this outbreak,with the prevalence as 11.95％.Cases mainly concentrated between 21 and 24,December,with curves noticed by pointsource and peaked on the 22nd.Symptoms mainly appeared as vomiting,with watery stool.Patients were founded in every village,with the highest prevalence appeared in the elderly (25.20％) but involved in every age group.Sapovirus infection was most frequently seen in the population under 25 years of age,with overall prevalence as 16.41％.People older than 50 years of age took the second place (12.05％),and the lowest was seen between 25 to 49 years age group (8.42％).Sapovirus was detected in both swab and fecal specimens,with the positive rate as 38.78％.Both general and heatresistant coliformed-bacteria were detected in the 24 monitored water samples,with the qualification rates as 95.83％,45.83％ and 50.00％,respectively.However,Sapovirus was not detected in any of the water samples.Conclusions Our findings confirmed that an outbreak,caused by sapovirus infection in CT community was most likely related to the central water supply.The surveillance system for infectious diarrheal diseases should be improved and both public health awareness and surveillance programs on drinking water should be strengthened.

AIM:To investigate the changes of cardiac structure and function in rats with type 2 diabetic melli-tus(T2DM),and to explore the mechanisms underlying diabetic cardiomyopathy.METHODS:The cardiac structure and function were measured by echocardiography in Zucker diabetic fatty(ZDF)rats and their control Zucker lean(ZL)rats. The size of the cardiomyocytes was determined by wheat germ agglutinin staining.The protein expression of atrial natriuretic peptide(ANP),β-myosin heavy chain(β-MHC), receptor for advanced glycation end products(RAGE), L-type cal-cium channel α1C subunit(CaV1.2)and Orai1 was assessed by Western blot.RESULTS:Compared with the ZL control rats,the thickness of left ventricular wall,ejection fraction(EF),fractional shortening(FS)and the sizes of cardiomyo-cytes were significantly increased,and diastolic function was decreased in the ZDF rats(P<0.05).The protein expression of β-MHC, ANP, RAGE and Orai1 was increased, while the expression of Ca V1.2 was decreased in ZDF rats(P <0.05).CONCLUSION:T2DM rats show the prominent features including cardiomyocyte hypertrophy,ventricular hyper-trophy and compensatory enhancement of cardiac function, and the Ca2+handling and increase in RAGE expression may play important roles in the processes.