MicroRNA-16-5p exacerbates sepsis by upregulating aerobic glycolysis via SIRT3-SDHA axis.

Sepsis is a life-threatening condition, and treatment for sepsis in clinic is often not available, partially due to insufficient understanding of the pathogenesis of sepsis. Extensive study to elucidate the pathogenesis is required to improve the clinical management and outcome of sepsis. In this study, we investigated the pathogenesis of sepsis using peripheral blood mononuclear cells (PBMCs) from septic patients and studied the underlying mechanism of miR-16-5p on aerobic glycolysis in lipopolysaccharide (LPS)-treated THP1 and Raw264.7 cells. The levels of RNA and protein were determined by real-time quantitative PCR and immunoblotting assay, respectively. The production of high mobility group box 1 (HMGB1) was measured by enzyme-linked immunosorbent assay. The levels of succinate and lactate were determined using colorimetric kits. The extracellular acidification rate (ECAR) and oxygen consumption rate (OCR) were measured by extracellular flux analyzer. The results showed that the expression of miR-16-5p was elevated, while sirtuin 3 (SIRT3) was decreased in PBMCs from septic patients and LPS-treated cells, along with accumulation of acetylated succinate dehydrogenase complex, subunit A. Concomitantly, an increase in HMGB1, succinate, lactate, as well as ECAR and a decrease in OCR were observed. Knockdown of miR-16-5p upregulated SIRT3 expression, facilitated SDHA deacetylation, and attenuated sepsis-related aerobic glycolysis. Further study identified that SIRT3 is targeted by miR-16-5p, and overexpression of SIRT3 rescued LPS-induced responses via deacetylation of SDHA. Our findings revealed a novel miR-16-5p-regulated SIRT3-SDHA axis in sepsis and provided novel insights for sepsis treatment.

Effects of antimicrobial peptides (AMPs) on blood biochemical parameters, antioxidase activity, and immune function in the common carp (Cyprinus carpio).

Antibiotic use in livestock feed additives has resulted in harmful residue accumulation and spread of drug-resistance. We examined the use of antimicrobial peptides (AMPs) as a safer alternative to antibiotics in feeding the common carp. AMPs were added to common carp basal diets (Control) as additives at four concentrations: 100 mg kg(-1) (B1), 200 mg kg(-1) (B2), 400 mg kg(-1) (B3), 600 mg kg(-1) (B4) by dry weight of basal diet. After a 60-day feeding experiment, the final weight, DG and SGR of carps on B1, B2 and B3 diet were significantly higher than the control (p < 0.05). The FCR of carps on B1, B2 and B3 diet were significantly lower than the control (p < 0.05). Carps on B2, B3, and B4 diets showed significantly lower (p < 0.05) levels of triglyceride than the control. B4-fed carps showed significantly lower (p < 0.05) levels of total protein, albumin, and total cholesterol than the control. However there was no remarkable difference (p > 0.05) in levels of uric ammonia, globulin, glutamic-pyruvic transaminase, glutamic-oxalacetic transaminase, lactic dehydrogenase and blood glucose in all groups. The serum superoxide dismutase and catalase activity of B1-fed carps was significantly higher (p < 0.05) than the control and B4-fed carps. The serum alkaline phosphate activity of carps on B1 diets was significantly higher (p < 0.05) than B4-fed carps. The serum acid phosphatase activity of B1-fed carps was significantly higher (p < 0.05) than the control and other antimicrobial peptide-fed groups. The serum lysozyme activity of carps on B1, B2, and B3 diets was significantly higher (p < 0.05) than the control- and B4-fed carps. Regarding immune factors in serum, the levels of immunoglobulin (Ig) and interleukin (IL)-1ß in B1-fed carps were significantly higher (p < 0.05) than the control and other groups, while IL-1α levels in B1-fed carps was significantly higher (p < 0.05) than the control-, B2-, and B3-fed carps. Furthermore, there were no significant differences in the content of MHC among the five groups. In conclusion, antimicrobial peptide can reduce triglyceride levels in serum, enrich oxidation resistance, and improve immunity of the common carp. It showed that appropriate concentration of antibacterial peptide as supplements in diets for common carp increased the final weight, DG, SGR and decreased the FCR.

Potential distribution of Leptotrombidium scutellare in Yunnan and Sichuan Provinces, China, and its association with mite-borne disease transmission.

BACKGROUND: Leptotrombidium scutellare is one of the six main vectors of scrub typhus in China and is a putative vector of hemorrhagic fever with renal syndrome (HFRS). This mite constitutes a large portion of the chigger mite community in southwest China. Although empirical data on its distribution are available for several investigated sites, knowledge of the species' association with human well-being and involvement in the prevalence of mite-borne diseases remains scarce. METHODS: Occurrence data on the chigger mite were obtained from 21 years (2001-2021) of field sampling. Using boosted regression tree (BRT) ecological models based on climate, land cover and elevation variables, we predicted the environmental suitability for L. scutellare in Yunnan and Sichuan Provinces. The potential distribution range and shifts in the study area for near-current and future scenarios were mapped and the scale of L. scutellare interacting with human activities was evaluated. We tested the explanatory power of the occurrence probability of L. scutellare on incidences of mite-borne diseases. RESULTS: Elevation and climate factors were the most important factors contributing to the prediction of the occurrence pattern of L. scutellare. The most suitable habitats for this mite species were mainly concentrated around high-elevation areas, with predictions for the future showing a trend towards a reduction. Human activity was negatively correlated with the environmental suitability of L. scutellare. The occurrence probability of L. scutellare in Yunnan Province had a strong explanatory power on the epidemic pattern of HFRS but not scrub typhus. CONCLUSIONS: Our results emphasize the exposure risks introduced by L. scutellare in the high-elevation areas of southwest China. Climate change may lead to a range contraction of this species towards areas of higher elevation and lessen the associated exposure risk. A comprehensive understanding of the transmission risk requires more surveillance efforts.

Probing cell membrane integrity using a histone-targeting protein nanocage displaying precisely positioned fluorophores.

Cell membrane integrity is fundamental to the normal activities of cells and is involved in both acute and chronic pathologies. Here, we report a probe for analyzing cell membrane integrity developed from a 9 nm-sized protein nanocage named Dps via fluorophore conjugation with high spatial precision to avoid self-quenching. The probe cannot enter normal live cells but can accumulate in dead or live cells with damaged membranes, which, interestingly, leads to weak cytoplasmic and strong nuclear staining. This differential staining is found attributed to the high affinity of Dps for histones rather than DNA, providing a staining mechanism different from those of known membrane exclusion probes (MEPs). Moreover, the Dps nanoprobe is larger in size and thus applies a more stringent criterion for identifying severe membrane damage than currently available MEPs. This study shows the potential of Dps as a new bioimaging platform for biological and medical analyses. Electronic Supplementary Material: Supplementary material (Figs. S1-S12 including distance information between neighboring fluorophores on Dps, TEM images, MALDI-TOF analysis, fluorescence spectra, confocal images, gel retardation analysis, tissue staining, and additional data) is available in the online version of this article at 10.1007/s12274-022-4785-5.

Human adenovirus (HAdV) infection in children with acute respiratory tract infections in Guangzhou, China, 2010-2021: a molecular epidemiology study.

BACKGROUND: Human adenovirus (HAdV) infection can cause a variety of diseases. It is a major pathogen of pediatric acute respiratory tract infections (ARIs) and can be life-threatening in younger children. We described the epidemiology and subtypes shifting of HAdV among children with ARI in Guangzhou, China. METHODS: We conducted a retrospective study of 161,079 children diagnosed with acute respiratory illness at the Guangzhou Women and Children's Medical Center between 2010 and 2021. HAdV specimens were detected by real-time PCR and the hexon gene was used for phylogenetic analysis. RESULTS: Before the COVID-19 outbreak in Guangzhou, the annual frequency of adenovirus infection detected during this period ranged from 3.92% to 13.58%, with an epidemic peak every four to five years. HAdV demonstrated a clear seasonal distribution, with the lowest positivity in March and peaking during summer (July or August) every year. A significant increase in HAdV cases was recorded for 2018 and 2019, which coincided with a shift in the dominant HAdV subtype from HAdV-3 to HAdV-7. The latter was associated with a more severe disease compared to HAdV-3. The average mortality proportion for children infected with HAdV from 2016 to 2019 was 0.38% but increased to 20% in severe cases. After COVID-19 emerged, HAdV cases dropped to 2.68%, suggesting that non-pharmaceutical interventions probably reduced the transmission of HAdV in the community. CONCLUSION: Our study provides the foundation for the understanding of the epidemiology of HAdV and its associated risks in children in Southern China.

[Observation of the clinical efficacy of the bone filling bag vertebroplasty for the treatment of osteoporotic vertebral compression fractures].

OBJECTIVE: To explore the clinical effects of bone filling bag vertebroplasty in treating osteoporotic vertebral compression fractures. METHODS: The clinical data of 127 patients (145 vertebrae) with osteoporotic vertebral compression fractures who corresponded the criteria of inclusion and exclusion from December 2015 to June 2017 were retrospectively analyzed. Responsible vertebral bodies were identified by clinical situation, X-rays, CT scan, MRI. Among them, 95 cases (110 vertebrae) were treated by percutaneons kyphoplasty (PKP group), there were 34 males (42 vertebrae) and 61 females (68 vertebrae), with an average age of (73.92±7.14) years, 47 thoracic vertebra (T8-T12) and 63 lumbar vertebra (L1-L5). Other 32 patients (35 vertebrae) were treated by bone filling bag vertebroplasty(bone filling bag vertebroplasty group). There were 11 males (12 vertebrae) and 21 females (23 vertebrae), with an average age of (71.56±7.89) years, 16 thoracic vertebra (T9-T12) and 19 lumbar vertebra(L1-L5). Postoperative pain after 3 days, vertebral body height, improvement of lumbar function were recorded and bone cement diffusion and leakage were observed by X-rays. RESULTS: All operations were successful and no complications were found. In bone filling bag vertebroplasty group, operation time was (31.75±4.99) min, postoperative VAS score at 3 days was(2.38±0.94) points, anterior and middle height of the vertebral body were(19.54±2.36) mm and (18.16±2.65) mm, respectively; ODI score was(25.19±5.49) points, all above items after operation were better than preoperation(P<0.01), but there was no significant difference between two groups(P>0.05). Bone cement was patchy, clumpy or slightly dispersed by X-rays at 3 days after operation, the leakage rate of bone cement in bone filling bag vertebroplasty group was 2.86%(1/35), while was 6.36%(18/110) in PKP group, all of them were "trailing sign", there was significant difference between two groups (P<0.05). CONCLUSIONS: The clinical effect of bone filling bag vertebroplasty in the treatment of osteoporotic vertebral compression fractures is similar to percutaneous kyphoplasty, it can effectively relieve the pain, restore part vertebral body height and obviously reduce the leakage rate, with safer, it is a simple, rapid and effective therapeutic method.

Antimicrobial resistance and prevalence of CvfB, SEK and SEQ genes among Staphylococcus aureus isolates from paediatric patients with bloodstream infections.

Staphylococcus aureus (S. aureus) is one of the most frequently isolated pathogens in neonatal cases of early and late-onset sepsis. Drug resistance profiles and carriage of toxin genes may affect the treatment and outcome of an infection. The present study aimed to determine the antimicrobial resistance patterns and frequencies of the toxin-associated genes conserved virulence factor B (CvfB), staphylococcal enterotoxin Q (SEQ) and staphylococcal enterotoxin K (SEK) among S. aureus isolates recovered from paediatric patients with bloodstream infections (BSIs) in Guangzhou (China). Of the 53 isolates, 43.4% were methicillin-resistant S. aureus (MRSA), and resistance rates to penicillin, erythromycin, clindamycin, trimethoprim/sulfamethoxazole, tetracycline, and ciprofloxacin of 92.5, 66.0, 62.3, 13.2, 20.8 and 1.9% were recorded, respectively. However, no resistance to nitrofurantoin, dalfopristin/quinupristin, rifampicin, gentamicin, linezolid or vancomycin was detected. Resistance to erythromycin, clindamycin and tetracycline in the MRSA group was significantly higher than that in the methicillin-susceptible S. aureus (MSSA) group. No significant differences in antimicrobial resistance patterns were noted between two age groups (≤1 year and >1 year). The proportion of S. aureus isolates positive for CvfB, SEQ and SEK was 100, 34.0 and 35.8%, respectively, with 24.5% (13/53) of strains carrying all three genes. Compared with those in MSSA isolates, the rates of SEK, SEQ and SEK + SEQ carriage among MRSA isolates were significantly higher. Correlations were identified between the carriage of SEQ, SEK and SEQ + SEK genes and MRSA (contingency coefficient 0.500, 0.416, 0.546, respectively; P<0.01). In conclusion, MRSA isolated from the blood of paediatric patients with BSIs not only exhibited higher rates of antimicrobial resistance than MSSA from the same source, but also more frequently harboured SEK and SEQ genes. The combination of the two aspects influenced the dissemination of MRSA among children. The present study clarified the characteristics of BSI-associated S. aureus and enhanced the current understanding of the pathogenicity and treatment of MRSA.

[Effects of different treatments on patients with osteoporotic fracture after percutaneous kyphoplasty].

OBJECTIVE: To evaluate effects of different treatments on patients with osteoporotic vertebral fracture after percutaneous kyphoplasty in pain and function. METHODS: From March 2010 to March 2012,138 patients (165 vertebrae) with thoracic and lumbar vertebral osteoporotic fracture were randomly divided into three groups (control group, treatment group and comprehensive group), 46 cases in each group, and all patients were treated by PKP. Control group were treated with calcium and calcitriol after operation, treatment group added salmon calcitonin see calcimar based on control group, comprehensive group added incrementality waist musculi dorsi function exercise based on treatment group. VAS, ODI scores and BMD before operation, 3 d, 2 weeks, 1 month, 6 months and 12 months after operation were detected and compared. RESULTS: All operation were performed successfully,38 cases (45 vertebrae) in control group, 36 cases (44 vertebrae) in treatment group and 40 cases (49 vertebrae) were obtained complete following up, there was no significant meaning in following time among three groups (P>0.05). Postoperative VAS and ODI scores at 3 d, 2 weeks and 1 month among three groups were lower than that of before operation (P<0.01). Compared with control group, postoperative VAS score at 3 d, 2 weeks and 1 month were decreasedin treatment group and comprehensive group, but there was no significant meaning in ODI scores (P>0.05). At 6 and 12 months after operation,there was no significant differences in VAS and ODI between control group and treatment group (P>0.05), while VAS score in comprehensive group decreased much than other two groups,decreased continuously (P<0.01). At 12 months after operation, BMD among three groups were increased more than preoperative,and BMD in comprehensive group was more obviously than that of in control and treatment group. CONCLUSION: PKP, an effective method for the treatment of thoracic and lumbar vertebral osteoporotic fracture, could improve short-term clinical effects by adding calcitonin with calcium supplements and activated vitamin D. Waist musculi dorsi function exercise could improve long-term clinical effects of PKP and improve quality of life.

[Comparative study of intertrochanteric fractures treated with proximal femur locking compress plate in aged].

OBJECTIVE: To compare the clinic outcomes of dynamic hip screw (DHS), intramedullary fixation (IF) and proximal femur locking plate (PF-LCP) in the treatment of intertrochanteric fractures in the elderly patients. METHODS: From July 2000 to August 2009, 165 old patients with intertrochanteric fractures were treated respectively by DHS, IF, PF-LCP. Fifty-eight patients were in DHS group including 30 males and 28 females with an average age of 71 years old; there were 30 cases of type II fracture of Jensen, 28 cases of type III fracture. Sixty-five patients were in IF group including 35 males and 30 females with an average age of 73 years old; there were 37 cases of type II fracture of Jensen, 28 cases of type III fracture. Forty-two patients were in PF-LCP group including 23 males and 19 females with an average age of 74 years old; there were 22 cases of type II fracture of Jensen,20 cases of type III fracture. The operative procedures,complications and therapeutic effects were compared among 3 groups. RESULTS: All patients were followed up from 15 to 21 months (averaged 18.3 months). The incision length and the operation time of IF group were shorter than that of DHS and PF-LCP, but there were no significant difference between DHS group and PF-LCP group. The intraoperattive blood loss, rehabilitation and healing time of IF and PF-LCP were less or shorter than that of DHS group, but there were no significant difference between IF group and PF-LCP group. The functional recovery of IF group and PF-LCP were better than that of DHS group, there were significant difference among 3 groups. The complications of PF-LCP group was fewer than that of IF group and DHS group. CONCLUSION: PF-LCP is the credible method for intertrochanteric fractures in the elderly patients, especially for severe comminuted fracture and osteoporosis, for it can reduce operation complications and benefit for fracture healing and hip functional recovery.

[Percutaneous kyphoplasty and conservative therapy for osteoporotic vertebral compression fractures: a clinical comparative study].

OBJECTIVE: To evaluate the clinical effects of percutaneous kyphoplasty (PKP) and conservative therapy in patients with osteoporotic vertebral compression fractures (OVCF). METHODS: The data of 63 patients with OVCF from Sep. 2007 to Apr. 2009 were retrospectively analyzed. There were 14 males and 49 females,ranging in age from 63 to 92 years, with an average of 73.4 years. Among them, 30 cases(38 vertebrae), 33 cases (35 vertebrae) were respectively treated with PKP, conservative therapy. The VAS score, the height of vertebral body and the neighboring vertebral fracture were observed during follow-up. RESULTS: All the patients were followed up from 10 to 15 months with an average of 13.3 months. Pain relieved in 27 cases with PKP, and VAS scores decreased from 8.32 before treatment to 2.63 at the 1st week after treatment; VAS scores still remained under 2 at the later follow-up. VAS scores had not changed at the 1st week after conservative therapy. VAS scores with conservative therapy were higher than with PKP after 1, 3 months (P < 0.05), but after 6 months, there was no significant difference between conservative therapy and PKP (P > 0.05). The average height of vertebral body on the X-rays increased in 4.1 mm at the 1st week after treatment with PKP (P < 0.01) and unchanged posteriorly. The height of vertebral body had some improvement at 3, 6 months after conservative therapy, but the height of vertebral body with PKP was significantly higher than with conservative therapy (P < 0.01). New fractures occurred in 4 cases (5 vertebrae) with PKP, in 2 cases (2 vertebrae) with conservative therapy. CONCLUSION: PKP is an effective method in treating osteoporotic vertebral compression fractures, which can relieve pain quickly, increase stability immediately, recover height of vertebral body, but maybe can increase the risk of new fracture. Conservative therapy is not without any merit, as long as systemic treatment can still make good prognosis.

[Clinical features and surgery in children with plastic bronchitis].

OBJECTIVE: To review the clinical features and therapeutic experience in children with plastic bronchitis. METHODS: Fourteen children with plastic bronchitis were reviewed retrospectively, 12 of which were under two years old. The clinical features are characterized by sudden onset, episodes of profound hypoxia and respiratory tract obstruction. SaO2 was between 0.70 and 0.80 even with mask oxygen inhalation. Eight cases were pyretic, 4 cases expectorated jel-like bronchial casts. The chest X-ray picture showed patchy consolidation or atelectasis unilaterally (10 cases) or bilaterally (2 cases). Pulmonary marking thickening and patchy shadow were observed in 2 cases. Twelve cases underwent rigid bronchoscopy and the bronchial casts were removed. Two cases underwent endotracheal intubation. RESULTS: Eight cases of 12 children received therapeutic bronchoscopy were cured. Other 4 cases had second therapeutic bronchoscopy and bronchial casts were removed again in 3 cases, one died from pulmonary hemorrhage. Two cases who underwent endotracheal intubation died from the multiple organ failure (MOF). Pathologic results showed:the bronchial casts were composed mainly of mucus and fibrin, inflammatory cell infiltrate were observed in 6 cases (Type 1, inflammatory), no cellular infiltrate occurred in 8 cases (Type 2, acellular). CONCLUSIONS: Plastic bronchitis is a severe and dangerous disease. The branching plastic casts may obstruct part or the entire tracheobronchial, causing respiratory failure. Bronchoscopy and pathologic examination are essential for it's diagnosis and treatment.