Restoration of constitutional alignment optimizes outcomes of computer navigated total knee arthroplasty: a prospective randomized controlled trial.

PURPOSE: The value of computer navigation in total knee arthroplasty (TKA) for arthritic knees continues to be debated. The purpose of this study was to evaluate the value of navigated TKA associated with updated alignment philosophy. METHODS: This prospective randomized controlled trial enrolled 38 consecutive patients (76 knees) and were randomly assigned to both groups. The demographic data and perioperative data were recorded. The coronal plane alignment of the knee (CPAK) classification was used to classify knee alignment phenotypes. Radiographic outcomes were measured and subgroup analysis was further performed. Clinical outcomes were evaluated using patient-reported outcome measures (PROMs). Surgery-related complications were recorded. RESULTS: The distribution of CPAK phenotypes following constitutional aligned TKA was equivalent to the native cohort, whereas the mechanical aligned TKA dramatically altered the phenotype distribution from type I and type II to type V and type IV. Final implant positioning was different between groups, with constitutional aligned TKA having larger cTCA (P = .004), joint line obliquity (P = .006), joint line distance (P = .033) and smaller sFCA (P = .013). Subgroup analysis showed higher actual accuracy of component positioning was achieved in navigated TKA, especially in knees with deformity of > 10° (P < .05). Patients reported higher HSS score at three months postoperatively in constitutional aligned group (P = .002). One patient in navigated group suffered femoral pin site fracture caused by a minor trauma. CONCLUSION: Computer navigated TKA allows for restoration of constitutional alignment and minimizes soft tissue release, which when compared to mechanical alignment may be associated with superior early outcomes.

Hydrogen sulfide protects against particle-induced inflammatory response and osteolysis via SIRT1 pathway in prosthesis loosening.

Wear debris-induced osteolysis and ensuing aseptic loosening is the main cause of implant failure and revision surgery. Wear debris-induced inflammatory response plays key roles in peri-implant osteolysis. Recently, substantial of evidence suggests that hydrogen sulfide (H2 S), the third gasotransmitter, is a critical player regulating inflammation. However, the role and therapeutic potential of H2 S in wear debris-induced inflammation and osteolysis remains to be defined. In the present study, we investigated the effect of H2 S on wear debris-induced pro-inflammatory cytokines expression and osteolysis in vitro and in vivo. With a slow-releasing H2 S donor GYY4137, our study demonstrated that H2 S attenuated wear debris-induced osteolysis and osteoclastogenesis in murine calvaria resorption models. The expression of tumor necrosis factor-alpha (TNF-α), interleukin-1ß (IL-1ß), and interleukin-6 (IL-6) that stimulated by wear particles were significantly reduced by GYY4137. Further, the level of sirtuin 1 (SIRT1), which possesses anti-inflammation property, was examined in vivo and in macrophages. And we found that wear debris decreased the expression of SIRT1. Cotreated macrophages with GYY4137 in part reversed the decline of SIRT1. More importantly, with the SIRT1 recombinant lentivirus and small interfering RNAs (siRNA) against SIRT1, our data indicated that SIRT1 mediated the inhibitory effects of GYY4137 on wear debris-induced inflammation. Collectively, these results suggested that exogenous H2 S production (via H2 S donors) may represent a potential approach for the treatment of wear particle-induced osteolysis.

Paeoniflorin inhibits activation of the IRAK1-NF-κB signaling pathway in peritoneal macrophages from lupus-prone MRL/lpr mice.

Systemic lupus erythematosus (SLE) is a chronic and multisystemic autoimmune disease. Interleukin-1 receptor-associated kinase 1 (IRAK1) is associated with the susceptibility of SLE in humans and paeoniflorin has recently been reported to exhibit immunosuppressive properties. The aim of this study was to determine the effect of paeoniflorin on lipopolysaccharide (LPS)-triggered macrophage activation and and its role in LPS-induced IRAK1-nuclear factor κB (NF-κB) signaling pathways. Peritoneal macrophages from lupus-prone MRL/lpr mice and ICR mice were isolated, prepared and cultured. Cells were treated with LPS alone or LPS with paeoniflorin, and macrophage proliferation was analyzed using the CCK8 assay. The expression of IRAK1 in cells was analyzed by immunofluorescence staining. The level of gene expression of IRAK1, NF-κB, tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) was measured by RT-PCR, and TNF-α, IL-6 levels in the cell supernatant were determined by ELISA. The protein expression of IRAK1 and downstream molecules tumor necrosis factor receptor-associated factor 6 (TRAF6), inhibitor of nuclear factor kappa-B kinase (IKK), NF-kappa-B inhibitor alpha (IKBα), and NF-κB was detected by Western-blot analysis. Paeoniflorin was found to decrease the phosphorylation of IRAK1 and its downstream proteins induced by LPS and inhibit the expression of TNF-α and IL-6. Taken together, the data obtained indicate that paeoniflorin inhibits LPS-induced cell activation by inhibiting the IRAK1-NF-κB pathway in MRL/lpr mouse macrophages. Therefore, paeoniflorin may be a potential therapy for SLE.

Direct differentiation of herbal medicine for volatile components by a multicapillary column with ion mobility spectrometry method.

In this work, a headspace system coupled to a gas chromatographic column and ion mobility spectrometry was applied as a screening system to differentiate the crude and processed "atractylodis macrocephalae rhizoma" samples. The obtained ion mobility data were consecutively processed by principal component analysis and Radar fingerprint chart methods. From the principal component analysis for the initial solution to original variables, the two principal components accounted for 68 and 13% of the total variance, respectively. The established method was proven to be valuable for classification, discrimination between herbal medicines from different processing procedures.

The causality between gut microbiota and ankylosing spondylitis: Insights from a bidirectional two-sample Mendelian randomization analysis.

BACKGROUND: The association between gut microbiota and ankylosing spondylitis (AS) has been reported in the literature; however, whether the two are correlative is unclear. METHODS: Single nucleotide polymorphisms associated with the gut microbiome composition and AS (968 AS cases and 336 191 controls) were obtained from published genome-wide association studies in this two-sample Mendelian randomization (MR) study. The causal relationship between gut microbiota and AS was estimated using the inverse-variance weighted method, and the robustness of our findings was confirmed through a comprehensive series of sensitivity analyses. RESULTS: Anaerotruncus (OR = 0.9984, 95% CI, 0.9968-0.9999, p = .0405) and Ruminococcaceae UCG002 (OR = 0.9989, 95% CI, 0.9979-0.9999, p = .0375) were protective against AS. Defluviitaleaceae (OR = 1.0015, 95% CI, 1.0005-1.0025, p = .0048), Butyricicoccus (OR = 1.0016, 95% CI, 1.0001-1.0032, p = .0429), Coprococcus 3 (OR = 1.0016, 95% CI, 1.0000-1.0032, p = .0463), and Defluviitaleaceae UCG011 (OR = 1.0016, 95% CI, 1.0005-1.0027, p = .0041) exhibited significant positive correlations with heightened susceptibility to AS. Reverse MR revealed that AS does not affect the gut microbial composition. CONCLUSION: Our study has established a genetically-based causal relationship between gut microbiota and AS. This finding suggests that we may be able to target and regulate specific bacterial groups in the gut to prevent and treat AS.

Mid-Term Outcomes of Navigation-Assisted Primary Total Knee Arthroplasty Using Adjusted Mechanical Alignment.

OBJECTIVE: The adjusted mechanical alignment (aMA) technique is an extension of conventional mechanical alignment (MA), which has rarely been reported. The purpose of this study was to evaluate mid-term outcomes of navigation-assisted total knee arthroplasty (TKA) using aMA. METHODS: This retrospective cohort study enrolled 63 consecutive patients (77 knees) who underwent navigation-assisted TKA using aMA between September 2017 and October 2019. Fifty-two consecutive patients (61 knees) who underwent TKA using MA during the same period were assessed as the controlled group. The demographic data and perioperative data were recorded. The parameters of resection and soft tissue balance including tibia resection angle, frontal femoral angle, axial femoral angle, joint line translation, medial and lateral gap in extension and flexion position were recorded. Radiographic parameters and functional scores including the Hospital for Special Surgery (HSS) score, Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score, and Forgotten Joint Score-12 (FJS-12) were evaluated. Surgery-related complications were recorded. The average follow-up was 3.5 years, with a minimum of 2.4 years. RESULTS: The frontal femoral angle was 2.55° ± 1.08° in aMA group versus 0.26° ± 0.60° in MA group (p < 0.001). The axial femoral angle was 3.07° ± 2.23° external in aMA group versus 2.30° ± 1.70° in MA group (p = 0.027). The lateral flexion gap was wider in the aMA group, with a mean of 0.71 mm more laxity (p = 0.001). Postoperative coronal alignment was 177.03° ± 1.82° in aMA group versus 178.14° ± 1.69° in MA group (p < 0.001). The coronal femoral component angle was 92.62° ± 2.78° in aMA group versus 90.85° ± 2.01° in MA group (p < 0.001). Both aMA-TKA and MA-TKA achieved satisfactory mid-term clinical outcomes. However, the HSS scores at 1 month postoperatively were significantly higher using aMA than using MA (p < 0.001). CONCLUSION: Navigation-assisted TKA using aMA technique obtained satisfactory mid-term clinical outcomes. The aMA technique aims to produce a biomimetic wider lateral flexion-extension gap and minimize releases of soft tissues, which might be associated with better early clinical outcomes than MA technique.

l-Lactic Acid Production via Sustainable Neutralizer-Free Route by Engineering Acid-Tolerant Yeast Pichia kudriavzevii.

l-Lactic acid (l-LA) is a platform chemical obtained via microbial fermentation at a near-neutral pH value. Large amounts of neutralizers are required during this process, which increases the production costs in downstream processing as well as environmental burden. To address this challenge, an acid-tolerant yeast Pichia kudriavzevii E1 was isolated and metabolically engineered to produce l-LA without neutralizers. The genome of strain E1 was sequenced and a CRISPR-Cas9 system was developed in this newly isolated strain. Subsequently, the gene encoding pyruvate decarboxylase (pdc) was knocked out to subdue ethanol formation. Furthermore, the l-lactate dehydrogenase gene from Weizmannia coagulans 2-6 and the codon-optimized L-ldhA gene from Bos taurus were introduced into P. kudriavzevii E1 chromosome to redirect the ethanol fermentation pathway to l-LA production. Deletion of the dld(chr3) gene further increased the optical purity of l-LA. After optimizing fermentation conditions, the maximum titer of l-LA in the 5 L fermenter reached 74.57 g/L without any neutralizers, with an optical purity of 100% and a maximum yield of 0.93 g/g glucose. This is the first report of optically pure l-LA production without neutralizers and the engineered acid-tolerant yeast paves the way for the sustainable production of l-LA via a green route.

Total Knee Arthroplasty Using Adjusted Restricted Kinematic Alignment for the Treatment of Severe Varus Deformity: Technical Note.

OBJECTIVE: To describe a new alignment technique of adjusted restricted kinematic alignment (arKA) for the treatment of severe varus deformity in total knee arthroplasty. METHODS: Three female patients (three severe varus knees) who underwent navigation-assisted total knee arthroplasty (TKA) using arKA from April 2020 to September 2020 were included in this study, with an average age of 71.33 years (range, 61 to 80 years). General anesthesia was given to all patients. Intraoperative observations including tibia resection angle, frontal femoral angle, axial femoral angle, medial and lateral gap in the extension and flexion positions and joint line translation were recorded. Also, operation duration and drainage volume were recorded. Radiographic parameters including the mechanical axis (α), coronal femoral component angle (ß), coronal tibial component angle (γ), sagittal femoral component angle (δ), tibial posterior slope angle (ε), femoral-patella angle (θ), and femoral notching were assessed. Clinical evaluation was performed using the Hospital for Special Surgery (HSS) Score and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) Score. Both individual and mean measurement data were displayed. RESULTS: The mean tibial resection was 4.00° varus (range, 3° to 5°), and the mean frontal femoral angle was 3.67° varus (range, 3° to 4°) in extension. The flexion lateral gap was wider than the medial gap with a mean laxity of 1.34 mm. Moreover, the mean axial femoral angle was 2.67° external (range, 0° to 6°) in flexion, and the mean joint line translation was 1.00 mm proximal (range, 0 to 3 mm). In addition, the mean preoperative mechanical axis was 156.22° (range, 153.65° to 158.90°) and the mean postoperative mechanical axis was 174.04° (range, 173.83° to 174.17°) with a mean correction of 17.82°. The mean femoral angle was 92.60° (range, 91.29° to 93.30°) and the mean tibial angle was 86.95° (range, 86.83° to 87.04°) in coronal plane. The HSS score improved from an average of 46.67 points (range, 42 to 51) preoperatively to 83.67 points (range, 81 to 86) at 3 months postoperatively. The mean WOMAC score was 16.33 points at 3 months postoperatively. CONCLUSIONS: The new alignment technique of arKA aims to balance the flexion and extension gap without extensive releases of soft tissue and restore the native pre-arthritic alignment, may be a promising alignment strategy for treating severe varus deformity. However, further study and comparison with other alignment techniques is needed.

[Midterm effectiveness of percutaneous compression plate for femoral neck fractures in young and middle-aged patients].

Objective: To analyze midterm effectiveness of percutaneous compression plate (PCCP) for femoral neck fractures in young and middle-aged patients. Methods: The clinical data of 173 young and middle-aged patients with femoral neck fractures who met the selection criteria and were treated with PCCP internal fixation between January 2011 and March 2019 were retrospectively analyzed. Among them, there were 102 males and 71 females; the age ranged from 18 to 59 years, with an average age of 44.9 years. The injury causes included falling in 112 cases, traffic accident in 32 cases, falling from height in 21 cases, struck in 7 cases, and sprain in 1 case. According to Garden classification, there were 90 cases without displacement (51 cases of type Ⅰ and 39 cases of type Ⅱ) and 83 cases with displacement (51 cases of type Ⅲ and 39 cases of type Ⅳ). According to Pauwels classification, there were 10 cases of type Ⅰ, 88 cases of type Ⅱ, and 75 cases of type Ⅲ. The time from injury to operation was 1-14 days, with an average of 4.5 days. The operation time, intraoperative blood loss, perioperative blood transfusion, and hospitalization stay were recorded; the fracture reduction was evaluated by Garden alignment index at 1 day after operation; fracture healing and complications were observed, and Harris score was used to evaluate the effectiveness at last follow-up. Results: The operation time was 34-130 minutes (mean, 78.6 minutes); the intraoperative blood loss was 10-250 mL (mean, 93.2 mL); 171 cases did not receive blood transfusion during perioperative period, 2 patients received blood transfusion of 400 mL and 800 mL respectively; the hospitalization stay was 3-19 days (mean, 11.3 days). All 173 cases were followed up 11-103 months, with an average of 42.6 months. Postoperative reduction quality was satisfactory in 170 cases and unsatisfactory in 3 cases. There were 13 cases of osteonecrosis of femoral head, 1 case of screw cutting out, 2 cases of screw withdrawal, 5 cases of femoral neck shortening, and no deep vein thrombosis in the lower extremity requiring surgical intervention. Fractures healed in 172 patients, and the healing time ranged from 3.0 to 7.5 months, with an average of 3.6 months; 1 case of nonunion occurred. Internal fixation was removed after fracture healing in 51 patients. At last follow-up, Harris score was excellent in 156 cases, good in 11 cases, fair in 3 cases, and poor in 3 cases, with an excellent and good rate of 96.5%. Conclusion: The treatment of femoral neck fractures with PCCP has advantages of rigid fixation, immediate weight-bearing, and sliding compression, reducing the incidences of osteonecrosis of femoral head and nonunion.

Transcriptomics and quantitative proteomics reveal changes after second stimulation of bone marrow-derived macrophages from lupus-prone MRL/lpr mice.

Innate immune memory can cause the occurrence and exacerbation of autoimmune diseases, and it is as well as being strongly associated with the pathogenesis of systemic lupus erythematosus (SLE), however, the specific mechanism remains to be further studied. We learned that IFN-γ stimulation generated innate immune memory in bone marrow-derived macrophages (BMDMs) and activated memory interferon-stimulated genes (ISGs). This research used IFN-γ and lipopolysaccharide (LPS) to treat BMDMs with lupus-prone MRL/lpr mice and showed that particular memory ISGs were substantially elevated in prestimulated macrophages. In order to identify the differentially expressed genes (DEGs), researchers turned to RNA-seq. GO and KEGG analysis showed that up-regulated DEGs were enriched in defense and innate immune responses, and were related to the expression of pattern recognition receptors (PRRs)-related pathways in macrophages. TMT-based proteome analysis revealed differentially expressed proteins (DEPs) up-regulated in BMDMs were abundant in metabolic pathways such as glucose metabolism. Our study found that after the secondary stimulation of MRL/lpr mice, the expression of PRRs in innate immune cells was changed, and IFN-related pathways were activated to release a large number of ISGs to promote the secondary response. At the same time, related metabolic modes such as glycolysis were enhanced, and epigenetic changes may occur. Therefore, SLE is brought on, maintained, and worsened by a variety of factors that work together to produce innate immune memory.

[Short-term effectiveness of reconstruction plate internal fixation via improved Stoppa approach combined with iliac fossa approach and Kocher-Langenbeck approach for complex acetabular fractures].

Objective: To investigate the short-term effectiveness of reconstruction plate internal fixation via improved Stoppa approach combined with iliac fossa approach and Kocher-Langenbeck approach for complex acetabular fractures. Methods: Between January 2015 and January 2020, 30 patients with complex acetabular fractures were treated with reconstruction plate internal fixation via improved Stoppa approach combined with iliac fossa approach and Kocher-Langenbeck approach. There were 20 males and 10 females with an average age of 52.1 years (range, 25-71 years). The cause of injury included traffic accident in 17 cases and falling from height in 13 cases. Among them, 14 cases were left acetabular fractures and 16 cases were right acetabular fractures. According to Letournel classification, there were 16 cases of double column fractures, 2 cases of transverse fracture with posterior wall fracture, 4 cases of anterior column and posterior hemi-transverse fractures, 8 cases of T-shaped fracture. The displacement distance of fracture ranged from 6 to 30 mm (mean, 11.6 mm). The time from injury to operation was 6-14 days (mean, 8.7 days). Results: The operation time was 2.0-4.5 hours (mean, 3.0 hours). The intraoperative blood loss was 200-800 mL (mean, 450.0 mL). All patients were treated with autologous blood transfusion during operation. All incisions healed by first intention after operation, and no infection occurred. All patients were followed up 12-15 months (mean, 13.4 months). The drainage tube was removed at 2-3 days after operation. After extubation, X-ray film and three-dimensional CT were performed to recheck the fracture reduction. According to Matta score system, 20 cases were excellent, 5 were good, and 5 were poor, the excellent and good rate was 83.3%. All fractures healed with the healing time of 16-25 weeks (mean, 17.7 weeks). According to Merle d'Aubigne-Postel score system, the hip function at 1 year after operation was rated as excellent in 18 cases, good in 6 cases, and fair in 6 cases, and the excellent and good rate was 80.0%. Two cases suffered from sciatic nerve injury due to traction during operation, 7 cases had heterotopic ossification, 2 cases had traumatic hip arthritis, and no other complications occurred. Conclusion: For complex acetabular fractures, the reconstruction plate internal fixation via improved Stoppa approach combined with iliac fossa approach and Kocher-Langenbeck approach can obtain good short-term effectiveness with good reduction and hip joint function, and less complications.

[Early effectiveness of computer navigation-assisted total knee arthroplasty].

OBJECTIVE: To estimate the early effectivenss of computer navigation-assisted total knee arthroplasty (TKA) by comparing with traditional TKA. METHODS: The clinical data of 89 patients (100 knees) underwent primary TKA between October 2017 and July 2018 were analyzed retrospectively, including 44 patients (50 knees) who completed the TKA under the computer-assisted navigation system as the navigation group and 45 patients (50 knees) treated with traditional TKA as the control group. There was no significant difference between the two groups ( P>0.05) in gender, age, body mass index, diagnosis, side, disease duration, Kellgren-Lawrence classification of osteoarthritis, and preoperative American Hospital for Special Surgery (HSS) score, range of motion (ROM), hip-knee-ankle angle (HKA) deviation. The operation time, incision length, difference in hemoglobin before and after operation, postoperative hospital stay, and the complications were recorded and compared between the two groups. The HSS score, ROM, and joint forgetting score (FJS-12) were used to evaluate knee joint function in all patients. Unilateral patients also underwent postoperative time of up and go test and short physical performance battery (SPPB) test. At 1 day after operation, the HKA, mechanical lateral distal femoral angle (mLDFA), mechanical medial proximal tibial angle (mMPTA), sagittal femoral component angle (sFCA), and sagittal tibial component angle (sTCA) were measured and calculated the difference between the above index and the target value (deviation); and the joint line convergence angle (JLCA) was also measured. RESULTS: The operations of the two groups were successfully completed, and the incisions healed by first intention. The operation time and incision length of the navigation group were longer than those of the control group ( P<0.05); the difference in difference of hemoglobin before and after the operation and the postoperative hospital stay between groups was not significant ( P>0.05). Patients in the two groups were followed up 27-40 months, with an average of 33.6 months. Posterior tibial vein thrombosis occurred in 1 case in each of the two groups, and 1 case in the control group experienced repeated knee joint swelling. The HSS scores of the two groups gradually increased after operation ( P<0.05); HSS scores in the navigation group at 1 and 2 years after operation, and knee ROM and FJS-12 scores at 2 years were significantly higher than those in the control group ( P<0.05). There was no significant difference in the postoperative time of up and go test and SPPB results between the two groups at 7 days after operation ( P>0.05); the postoperative time of up and go test of the navigation group was shorter than that of the control group at 2 years ( t=-2.226, P=0.029), but there was no significant difference in SPPB ( t=0.429, P=0.669). X-ray film measurement at 1 day after operation showed that the deviation of HKA after TKA in the navigation group was smaller than that of the control group ( t=-7.392, P=0.000); among them, the HKA deviations of 50 knees (100%) in the navigation group and 36 knees (72%) in the control group were less than 3°, showing significant difference between the two groups ( χ 2=16.279, P=0.000). The JLCA and the deviations of mLDFA, mMPTA, sFCA, and sTCA in the navigation group were smaller than those in the control group ( P<0.05). CONCLUSION: Compared with traditional TKA, computer navigation-assisted TKA can obtain more accurate prosthesis implantation position and lower limb force line and better early effectiveness. But there is a certain learning curve, and the operation time and incision length would be extended in the early stage of technology application.

Jieduquyuziyin Prescription Suppresses Inflammatory Activity of MRL/lpr Mice and Their Bone Marrow-Derived Macrophages via Inhibiting Expression of IRAK1-NF-κB Signaling Pathway.

Jieduquyuziyin prescription (JP) has been used to treat systemic lupus erythematosus (SLE). Although the effectiveness of JP in the treatment of SLE has been clinically proven, the underlying mechanisms have yet to be completely understood. We observed the therapeutic actions of JP in MRL/lpr mice and their bone marrow-derived macrophages (BMDMs) and the potential mechanism of their inhibition of inflammatory activity. To estimate the effect of JP on suppressing inflammatory activity, BMDMs of MRL/lpr and MRL/MP mice were treated with JP-treated serum, and MRL/lpr mice were treated by JP for 8 weeks. Among them, JP and its treated serum were subjected to quality control, and BMDMs were separated and identified. The results showed that in the JP group of BMDMs stimulated by Lipopolysaccharide (LPS) in MRL/lpr mice, the secretion of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) reduced, and the expressions of Interleukin-1 receptor-associated kinase 1 (IRAK1) and its downstream nuclear factor κB (NF-κB) pathway decreased. Meanwhile, the alleviation of renal pathological damage, the decrease of urinary protein and serum anti-dsDNA contents, the inhibition of TNF-α level, and then the suppression of the IRAK1-NF-κB inflammatory signaling in the spleen and kidney, confirmed that the therapeutic effect of JP. These results demonstrated that JP could inhibit the inflammatory activity of MRL/lpr mice and their BMDMs by suppressing the activation of IRAK1-NF-κB signaling and was supposed to be a good choice for the treatment of SLE.

Network Analysis of Depression-Related Transcriptomic Profiles.

Major depressive disorder is a common debilitating disorder that is associated with increased morbidity and mortality. However, the molecular mechanism underlying depression remains largely unknown. The current study investigated the association of depression with blood gene expression using data from the Alzheimer's Disease Neuroimaging Initiative (ADNI). Depression was measured by the geriatric depression scale, and the blood gene expression was measured by the Affymetrix Human Genome U219 Array. Linear regression was used to test the association between gene expression and depression, and the model was adjusted for age and sex. A total of 671 participants were included in our study (mean age 75 ± 8 years, 43.2% women). We found three genes were associated with depression, including COL1A2 (P = 8.9 × 10-8), RNF150 (P = 1.4 × 10-7) and CTGF (P = 8.3 × 10-7). An interaction network was built, and the pathway analysis indicated that many depression-related genes were involved in the neurotrophin signaling pathway (P = 2.1 × 10-7). Future studies are necessary to validate our findings and further investigate potential mechanism of depression.

Jieduquyuziyin Prescription-Treated Rat Serum Suppresses Activation of Peritoneal Macrophages in MRL/Lpr Lupus Mice by Inhibiting IRAK1 Signaling Pathway.

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease, and Jieduquyuziyin prescription (JP) is a traditional Chinese medicine (TCM) formula that has been testified to be effective for SLE treatment as an approved hospital prescription for many years in China. However, its mechanism of action in the treatment of this disease is largely unknown. The purpose of this study was to determine whether JP-treated rat serum can inhibit the activation of peritoneal macrophages in MRL/lpr mice by downregulating the IRAK1 signaling pathway, thereby achieving the effect of improving SLE. The JP-treated rat serum was prepared, and the peritoneal macrophages of MRL/lpr lupus mice were isolated in vitro, and the effect of JP on cell viability was detected by the CCK8 method. After LPS induction and shRNA lentiviral transfection, the effect of JP on the expression of IRAK1 in cells was detected by immunofluorescence staining. The content of TNF-α and IL-6 in the cell supernatant was determined by ELISA. The expression of IRAK1, NF-κB, TNF-α, and IL-6 mRNA was detected by RT-PCR, and the expression levels of IRAK1, p-IRAK1, TRAF6, IKBα, p-IKBα, IKK + IKK, NF-κB, and p-NF-κB proteins was detected by western blot method. We investigated the role of JP in peritoneal macrophages of the MRL/lpr mouse and identified the possible mechanisms of action. The results showed that JP could reduce the phosphorylation of IRAK1 and its downstream proteins induced by LPS and inhibit the expression of inflammatory cytokines, including TNF-α and IL-6. In addition, after the transfection of cells with shRNA lentiviral, the results of JP tended to be consistent. In conclusion, JP may inhibit the activation of peritoneal macrophages in MRL/lpr mice by downregulating the IRAK1-NF-κB signaling pathway, and IRAK1 may be a potential target for JP treatment of SLE.

Effect of Jieduquyuziyin prescription-treated rat serum on MeCP2 gene expression in Jurkat T cells.

How genomic DNA methylation and methyl CpG-binding protein 2 (MeCP2) gene expression affect the pathogenesis of systemic lupus erythematosus (SLE) remains poorly understood. Traditional Chinese medicine has a unique effect in the treatment of SLE patients. This study aimed to investigate the effect of Jieduquyuziyin prescription (JP)-treated rat serum on the gene expression of MeCP2 in Jurkat T cells and its role in the pathogenesis of SLE. Jurkat T cells were harvested, and drug-containing serum was prepared. The ferulic acid and paeoniflorin content in the drug-containing serum were determined by liquid chromatography-mass spectrometry (LC-MS/MS). 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) assays were used to screen the optimal concentration of drug-containing serum. The DNA methylation level in Jurkat T cells was detected with a Methylamp™ Total DNA Methylation Kit. The methylation status of the MeCP2 promoter region was detected using bisulfite modification and methylation-specific PCR (MSP). Real-time PCR was used to measure MeCP2 mRNA expression. Western blotting and flow cytometry were done to detect MeCP2 protein expression in Jurkat cell nuclei. Paeoniflorin and ferulic acid were detected in the drug-containing serum of JP-treated rats. The results showed that cell growth was affected in the high serum-containing drug group. The experimental results showed that JP and prednisone acetate increased the level of genomic DNA methylation and MeCP2 gene promoter region methylation in Jurkat cells. MeCP2 mRNA and protein levels were also increased in the JP and prednisone acetate groups. Furthermore, flow cytometry revealed that the expression of MeCP2 protein in Jurkat T cell nuclei was higher in the drug group than the blank control group, and these results were consistent with the western blot analysis results. Our study found that there is a negative correlation between drug-containing serum and cell survival rate. JP upregulated the levels of DNA methylation, MeCP2 mRNA and protein as effectively as prednisone acetate and thus may activate the MeCP2 gene by increasing the methylation level, thereby inhibiting the pathogenesis of SLE. Therefore, JP may potentially be used to treat SLE patients. The Jurkat T lymphocyte in vitro experiments provided a foundation to study the effects of JP on the lupus mouse CD4+ T cell methylation mechanism and to further explore the pathogenesis of SLE.

Integrated DNA and RNA extraction using magnetic beads from viral pathogens causing acute respiratory infections.

Current extraction methods often extract DNA and RNA separately, and few methods are capable of co-extracting DNA and RNA from sputum. We established a nucleic acid co-extraction method from sputum based on magnetic beads and optimized the method by evaluating influencing factors, such as the guanidinium thiocyanate (GTC) and dithiothreitol (DTT) concentrations, magnetic bead amount, incubation temperature, lysis buffer pH and RNA carrier type. The feasibility of the simultaneous nucleic acid co-extraction method was evaluated by amplifying DNA and RNA viruses from a single clinical specimen with a multiplex RT-qPCR method. Both DNA and RNA were most efficiently extracted when the GTC and DTT concentrations were 2.0 M and 80 mM, respectively, 20 µl magnetic beads were added, the incubation temperature was 80 °C, the pH was 8 or 9, and RNA carrier A was used. Therefore, we established a simple method to extract nucleic acids from two important respiratory viruses compared with other commercial kits. This magnetic beads-based co-extraction method for sputum followed by a multiplex RT-qPCR can rapidly and precisely detect DNA and RNA viruses from a single clinical specimen and has many advantages, such as decreased time, low cost, and a lack of harmful chemicals.

Treatment of femoral neck fracture with percutaneous compression plate: preliminary results in 74 patients.

OBJECTIVE: The purpose of this study was to present the surgical technique and clinical results of percutaneous compression plate (PCCP) for the treatment of femoral neck fractures. METHODS: Between December 2010 and April 2013, 74 consecutive patients with 74 femoral neck fractures were treated by closed reduction and PCCP implants in our university hospital. Their mean age was 51.3 years (range, 15-83 years); 38 (51.4%) were male and 46 (62.2%) of the fractures were on the left. The patients' clinical and radiographic data were analyzed retrospectively. RESULTS: There were 45 undisplaced (60.8%) and 29 displaced fractures (39.2%). Eight patients (10.8%) were lost to follow-up. The mean duration of follow-up was 18.8 months for the remaining 66 patients. At the last follow-up, mean Harris hip score was 92.9 (range, 75-100), and 65 patients (98.5%) had excellent and good outcomes. Sixty-five patients (98.5%) were able to walk independently and one (1.5%) with walking-sticks. The mean time to clinical fracture healing was 3.9 months. There were no cases of nonunion. Two patients (3.0%) had delayed union and two (3.0%) developed avascular necrosis, one of 29 (3.7%) with a displaced fracture and one of 45 (2.6%) with an undisplaced fracture. There were no other complications or prosthetic replacement. CONCLUSIONS: PCCP is a stable internal fixation device that resists axial and rotational stresses. Our PCCP procedure has a low incidence of nonunion and avascular necrosis.