Evaluate the Outcomes of Senile Femoral Intertrochanteric Fractures Treated with Proximal Femoral Nail Anti-Rotation Internal Fixation to Those Treated with Prosthetic Femoral Head Replacement.

Objective: This study aims to evaluate the outcomes of senile femoral intertrochanteric fractures treated with proximal femoral nail anti-rotation (PFNA) internal fixation to those treated with prosthetic femoral head replacement. Methods: A total of 100 elderly patients suffering from femoral intertrochanteric fracture were selected for the study. They were divided into two groups (n = 50 in each group) based on fracture condition and preferred treatment. We compared perioperative indexes, complications, Soluble cell adhesion molecules-1 (sICAM-1), and TGF-1 levels, and assessed hip function using the Harris hip score (Harris) at 3, 6, and 12 months after surgery in two groups of patients. Results: Although the study group had shorter operating times and less intraoperative bleeding than the control group (P < .05), they had longer hospital stays and required more time before returning to full weight-bearing after surgery (P < .05). Neither group had a higher or lower rate of problems than the other (P > .05). Patients' sICAM-1 and TGF-1 levels were not significantly different from one another before surgery (P > .05), but after surgery, the sICAM-1 level in the control group was lower than that in the study group, and the TGF-1 level was higher than that in the study group (P < .05). Conclusion: PFNA internal fixation treatment offers the advantages of short operation time and low intraoperative bleeding, ensuring surgical safety. However, it requires a longer bed rest time post-operation and extended full weight-bearing time, although long-term hip recovery is preferable.

Identification of the candidate genes of diagnosing rheumatoid arthritis using the single-cell sequencing technology and T cell subclusters analysis of patients with rheumatoid arthritis.

Objectives: This study aims to analyze the heterogeneity among different cell types in peripheral blood mononuclear cells (PBMC) in rheumatoid arthritis (RA) patients and to analyze T cell subsets to obtain key genes that may lead to RA. Materials and methods: The sequencing data of 10,483 cells were obtained from the GEO data platform. The data were filtered and normalized initially and, then, principal component analysis (PCA) and t-Distributed Stochastic Neighbor Embedding (TSNE) cluster analysis were performed using the Seurat package in R language to group the cells, thereby obtaining the T cells. The T cells were subjected to subcluster analysis. The differentially expressed genes (DEGs) in T cell subclusters were obtained, and the hub genes were determined by Gene Ontology (GO) functional enrichment analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis and protein-protein interaction (PPI) network construction. Finally, the hub genes were validated using other datasets in the GEO data platform. Results: The PBMC of RA patients were mainly divided into T cells, natural killer (NK) cells, B cells, and monocyte cells. The number of T cells was 4,483, which were further divided into seven clusters. The pseudotime trajectory analysis showed that the differentiation of T cells developed from cluster 0 and cluster 1 to cluster 5 and cluster 6. Through GO, KEGG and PPI analysis, the hub genes were identified. After validation by external data sets, nine genes were identified as candidate genes highly associated with the occurrence of RA, including CD8A, CCL5, GZMB, NKG7, PRF1, GZMH, CCR7, GZMK, and GZMA. Conclusion: Based on single-cell sequencing analysis, we identified nine candidate genes for diagnosing RA, and validated their diagnostic value for RA patients. Our findings may provide new sights for the diagnosis and treatment of RA.

Aberrant Methylation of miR-34b and IL-12B mRNA Promoters Contributes to the Reduced Severity of Ankylosing Spondylitis.

DNA methylation of Interleukin-12B (IL-12B) and miR-34b was proved to affect the expression of IL-12B and miR-34b, which were found to be involved in the pathogenesis of ankylosing spondylitis (AS). However, the molecular mechanisms underlying the role of IL-12B and miR-34b in AS remain to be explored. AS patients were divided into four groups according to their status of DNA methylation of miR-34b and IL-12B by bisulfite sequencing: HYPER-miR-34b + HYPO-IL-12B, HYPER-miR-34b + HYPER-IL-12B, HYPO-miR-34b + HYPER-IL-12B and HYPO-miR-34b + HYPO-IL-12B groups. Functional indicators were examined for patients with different status of DNA methylation in their miR-34b and IL-12B promoters. QPCR was performed to examine the expression of miR-34b and IL-12B mRNA under different conditions. ELISA was used to measure the expression of IL-12B p40 in the peripheral blood. Western blot was used to analyze the expression of IL-12B proteins. Luciferase assay was carried out to explore the suppressive role of miR-34b in IL-12B expression. The level of Ankylosing Spondylitis Disease Activity Score with C-reactive protein (ASDAS-CRP) was gradually increased in HYPER-miR-34b + HYPO-IL-12B,HYPER-miR-34b + HYPER-IL-12B,HYPO-miR-34b + HYPER-IL-12B and HYPO-miR-34b + HYPO-IL-12B groups, whereas the levels of Bath Ankylosing Spondylitis Functional Index (BASFI) and Bath Ankylosing Spondylitis Metrology Index (BASMI) were significantly elevated in the HYPO-miR-34b + HYPO-IL-12B group and diminished in the HYPER-miR-34b + HYPO-IL-12B group. The expression of miR-34b in the PBMCs and peripheral blood was remarkably higher in the HYPER-miR-34b + HYPO-IL-12B and HYPER-miR-34b + HYPER-IL-12B groups, whereas the expression of IL-12B was gradually decreased in the HYPER-miR-34b + HYPO-IL-12B, HYPER-miR-34b + HYPER-IL-12B, HYPO-miR-34b + HYPER-IL-12B and HYPO-miR-34b + HYPO-IL-12B groups. Luciferase assays with the transfection of miR-34b precursors suggested that miR-34b strongly suppressed the expression of IL-12B in THP-1 cells. In conclusion, our study demonstrated that hypermethylated miR-34b promoter led to evident upregulation of miR-34b, thus inhibiting the expression of IL-12B and alleviated the severity of ankylosing spondylitis by reducing the levels of factors including ASDAS-CRP, BASFI and BASMI.

Gut microbiota-dependent Trimethylamine N-Oxide are related with hip fracture in postmenopausal women: a matched case-control study.

The study evaluates the serum levels of Trimethylamine N-Oxide (TMAO), a gut microbial metabolite, in 286 postmenopausal women with hip fracture. From January 1, 2018 to December 31, 2018, eligible patients were included. Same women without fracture mated age were enrolled. TMAO serum levels were tested by ultra-high-performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). The serum levels of TMAO were significantly higher in patients with hip fracture than in those controls (P<0.001). The serum levels of TMAO were also higher in patients with hip fracture only than in those who also had upper limb fracture (P=0.001). High level of TMAO was proved a predictor of both hip fracture and had upper limb fracture combined hip fracture, after the adjustment of other existing risk factors [e.g., for each 1 uM increase of TMAO, odd ratio 1.16 (95% CI, 1.07-1.25), P < 0.001; and 1.12 (95% CI, 1.03-1.26), P=0.008, respectively]. In summary, increased TMAO serum levels associated with high risk of hip fracture, suggesting that increase TMAO may contribute to osteoporosis and fracture in postmenopausal women.

Low 25(OH) D serum levels are related with hip fracture in postmenopausal women: a matched case-control study.

PURPOSE: There is limited information on the prevalence of vitamin D deficiency among patients diagnosed with hip fracture in the Chinese Han population. Therefore, the aim of this study was to assess the effects of change in the serum levels of 25-hydroxyvitamin D [25(OH) D] and intact parathyroid hormone (iPTH) among postmenopausal women in North China with confirmed hip fracture. METHODS: This study was done from May 1, 2012 to April 30, 2014. Three hundred and forty-nine postmenopausal women who were diagnosed with first-ever hip fracture and 349 matched controls without fracture were used for this study. The 25(OH) D, iPTH, alkaline phosphatase, calcium, and phosphorus levels were measured in fasting venous blood samples collected from the subjects. A predesigned questionnaire was used to collect information on covariates for multivariate analyses to evaluate the hypothesized relationship between vitamin D deficiency and fracture risk. RESULTS: The serum 25(OH) D levels were found to be significantly (P < 0.0001) lower in hip fracture patients than in the controls [37.0 (interquartile range [IQR] 28.0-48.0) nmol/L vs. 41.3 (IQR 32.0-54.5) nmol/L; P < 0.0001], and the iPTH levels were significantly higher in the former group [10.2 (IQR 6.3-14.9) pmol/L vs. 5.8 (IQR 4.1-6.6) pmol/L; P < 0.0001]. Further, a 25(OH) D level ≤50 nmol/L was found to independently indicate the occurrence of hip fracture [odds ratio (OR), 3.023; 95 % confidence interval (CI) 2.154-4.298], as well as hip fracture with concomitant upper limb fracture (OR 4.473; 95 % CI 2.984-10.532). Similarly, a serum iPTH level ≥6.8 pmol/L independently indicated the development of hip fracture (OR 2.498; 95 % CI 1.764-3.942), as well as hip fracture with concomitant upper limb fracture (OR 3.254; 95 % CI 1.998-7.984). CONCLUSIONS: Vitamin D insufficiency and secondary hyperparathyroidism were found to be common problems in the sample of postmenopausal women who had experienced hip fracture. Monitoring the alterations in the serum levels of 25(OH) D and iPTH could be applied clinically as independent risk factors for hip fracture.

[A preliminary study on repairing defects at medial malleolus in children by vascularized fibular head composite flap].

OBJECTIVE: To investigate the operative method and effectiveness of repairing defects at medial malleolus in children with vascularized fibular head composite flap. METHODS: Between November 2008 and January 2011, 8 children with bone and soft tissue defects at the medial malleolus were treated. There were 5 boys and 3 girls, aged 2-9 years (mean, 4.6 years). Injuries were caused by machine twisting in 2 cases and by wheel twisting in 6 cases. Soft tissue defect area ranged from 3.5 cm x 3.0 cm to 7.0 cm x 4.5 cm; defect was total in all medial malleolus. The disease duration from injury to admission was 2-8 hours (mean, 4.5 hours). Defects were repaired with vascularized fibular head composite flap carrying the skin around the head of the fibula in 5 cases, and with vascularized fibular head composite flap and skin flap above the medial malleolus in 3 cases having too large defect (> 5 cm x 4 cm). The donor sites were repaired with direct suture in 2 cases and with skin graft in 6 cases. RESULTS: All 8 fibular head composite flaps and 3 skin flaps above the medial malleolus survived completely. Wounds healed by first intention; the skin grafts at donor sites survived in the other cases except 1 case having local necrosis, with healing of incision by first intention. The patients were followed up 10 months to 3 years (mean, 22 months). The color and elasticity of the flaps were good. All the children had equal leg length. Of 8 cases, 6 had no joint valgus; 2 cases had progressive ankle varus after 1 year of operation. The ankle flexion and extension function returned to normal in 5 cases, and was slightly limited in 3 cases; horizontal side, forward and backward movements had no difference compared with normal side. According to American Orthopaedic Foot and Ankle Society (AOFAS) ankle function evaluation criteria, the results were excellent in 5 cases, and good in 3 cases at 10 months after operation. X-ray film showed that the ankle hole gap development of both sides was similar; no premature closure of the epiphysis or bone bridge formation of the medial malleolus was observed in 6 cases, and bone bridge formed in 2 cases after 1 year of operation. CONCLUSION: The satisfactory short-term effectiveness can be obtained in repairing children medial malleolus and soft tissue defects by vascularized fibular head composite flap, and the reconstructed medial malleolus can develop with the growth of children. Long-term effectiveness still need more follow-up study.

[Long-term regulation of arterial pressure and essential hypertension--II. Essential hypertension].

Although the mechanisms underlying the development of essential hypertension remain elusive, many observations point to the kidney as a primary actor and sodium as the main culprit (external factor) for development of hypertension. Dietary sodium has been existed for several thousands years in human being and it seems to be a civilized food habit. However, over the last few decades, experimental, observational and clinical data have continuously indicated that excess salt intake is positively associated with elevated blood pressure. It was also found that aged people is easier to be suffered from essential hypertension. By now, essential hypertension is frequently considered as a "civilized" disease, a disease of the kidney and the disease frequently occurring in the aged people. In the present mini review these features of essential hypertension are discussed in more details.

[Long-term regulation of arterial pressure and essential hypertension--I. Long-term regulation of arterial pressure].

Arterial blood pressure is regulated not by a single pressure controlling mechanism but instead by several interrelated mechanisms, each of which performs a specific function. By now, it is clear that kidney plays a dominant role in long-term regulation of arterial pressure. It is found that urine volume output markedly increases as the arterial blood pressure rises. It is the phenomenon of pressure-diuresis. Arterial blood pressure can be kept constantly by the action of pressure-diuresis when the excess accumulation of extracellular fluid in the body occurs. During this period of time kidney excretes a larger amount of urine volume. It is thus that under the conditions of the excess accumulation of extracellular fluid in the body, high level of the arterial blood pressure can only be observed as the renal function is abnormal.

[Organotypic cerebral slice culture and the course of neuronal development in it].

OBJECTIVE: To establish a method of organotypic cerebral culture. So as to pave the way for building some neurodegenerative disease models. METHODS: Organotypic cerebral cultures were prepared from prefrontal brain of neonatal SD rats. After culturing 7 to 14 days, 3 weeks, 4 weeks and 8 weeks, respectively, cerebral slices were fixed, dehydrated and sectioned in cryostat. The sections proceeded with Nissl staining and neurofilament high molecular weight (NFH) immunohistochemical staining. The difference was observed between controls and cultured slices using normal rats as controls. RESULTS: Nissl staining showed that pyramidal neurons in cultured slices were increased in volume and lightened in staining. The delaminating construction was clear from 1 to 4 weeks after culturing. In cultured slices, immunohistochemical staining showed that NFH positive pyramidal cells appeared on layer V on the tenth day and on both layers V and III after culturing 12 days. In the control group, NFH positive pyramidal cells appeared on layer V in 5-day-old rats, and appeared on both layers V and III in over 3-week-old rats. In cultured cerebral slices, the number of pyramidal neurons on layer V in M1 area was invariable from 12 days to 2 months. CONCLUSION: Orgaotypic cerebral culture can be used to study postnatal development for neocortex and build some in vitro models for neurodegenerative diseases.

[Establishment of the organotypic model of amyotrophic lateral sclerosis from the SD rats' spinal cord].

OBJECTIVE: To establish an in vitro model of amyotrophic lateral sclerosis (ALS) from the organotypic culture of SD rats' lumber spinal cord induced by the mitochondrial inhibitor,malonate sodium. METHOD: The lumber spinal cord prepared from the 6-day-old SD rats was cut into 350 microm coronarily, cultured on the Millicell-CM inserts which make the spinal cord culturing on the interface between air and fluid. First, the optimum malonate sodium dose was determined by adding different doses into the medium and counting the living motor neuron numbers by using immuno-histochemistry staining. Second, the ALS model was established as following: the cultures were divided into the malonate groups and the control groups, adding 2 mmol/L sodium malonate into the medium of the malonate groups an the 3rd day, continue culture to 12 days with this concentration; the control groups culturing without malonate. RESULTS: The organotypic characteristics are still kept till the end of the curlturing. After adding the sodium malonate, counting the number of motor neurons and interneurons on the different spinal slices in the different groups, the number of motor neuron in the cultured spinal cord was less than control (11.00+/-2.45 vs 15.29+/-1.70 per semislice at the end of the culturing, P<0.01), but the difference of the interneuron was not significant. CONCLUSION: The amyotrophic lateral sclerosis model is successful with selective injury of motor neuron, and this model can be used for the exploring of the theraptic method and its pathogenesis.

A novel PPAR alpha/gamma dual agonist inhibits cell growth and induces apoptosis in human glioblastoma T98G cells.

AIM: To examine the effect of a novel peroxisome proliferator-activated receptor (PPAR) alpha/gamma dual agonist TZD18 on cell proliferation and apoptosis in human glioblastoma T98G cells and its possible mechanism. METHODS: RT-PCR, MTT, TUNEL, Flow cytometry, and Western blot analysis were employed. RESULTS: TZD18 inhibited the growth of T98G cells in a concentration-dependent manner, which was associated with a G1 to S cell cycle arrest. Besides, significant apoptosis was induced after treatment with a non-toxic dose of TZD18. During the process, the expression of Bcl-2 protein was down-regulated, while that of Bax and p27kip proteins was up-regulated, and the activity of caspase-3 was elevated. However, this effect appeared to be PPARalpha and PPARgamma independent since their antagonists could not reverse this effect. CONCLUSIONS: TZD18, a novel PPARalpha/gamma dual agonist, inhibited cell growth and induce apoptosis in human glioblastoma T98G cells in vitro, indicating a therapeutic potential for TZD18 in the treatment of glioblastoma.

[Ion channels on T lymphocyte].

There is increasing evidence to show that ion channels on lymphocytes play a very important role in the regulation of immune functions. In T lymphocytes, there are three types of ion channels on cell membrane: Ca2+, K+ and Cl- channel. The influx of Ca2+ into T lymphocyte through Ca2+ channel (CRAC) may act as a second messenger to activate T lymphocyte when antigen binds to the receptor (TCR). The efflux of K+ from T lymphocyte through the K+ channel contributes to the formation of T cell membrane potential. The level of the membrane potential may affect the influx of Ca2+ into T cells. Therefore, the activation and the functions of T cell can be regulated by K+ channel indirectly. Cl- channel in T lymphocyte was found in recent years and it is probably involved in the regulation of cell volume. The recent progress on ion channels in T lymphocyte is summarized briefly in the present paper.

[Integer multiple rhythm in the spontaneous beating rhythm of cardiac myocytes and its possible mechanism].

OBJECTIVE: To introduce the characteristics of the integer multiple rhythm of cultured cardiac myocytes and to explore the cause of its generation. METHOD: Spontaneous beating rhythms of cultured cardiac myocytes were observed with photometry system and stochastic Chay model was used to simulate the experimental results. RESULT: Integer multiple rhythm was observed in the experiment. This kind of rhythm is similar to phenomena of sinus arrest. The integer multiple rhythm similar to that of the experiments was simulated in stochastic Chay model, and was demonstrated to be induced by the mechanism of autonomous stochastic resonance. CONCLUSION: The integer multiple rhythm observed in the experiment might be generated via the effect of autonomous stochastic resonance.

A protein in pig spleen similar to immune suppressive protein of stress.

AIM: To purify a protein in pig spleens, which was similar to immune suppressive protein of stress (ISPS), and characterize its properties and functions. METHODS: 1) Pig spleen was extracted in dilute hydrochloric acid. 2) The extract was ultra-filtrated for having high molecular weight proteins (Mr>30 000). 3) The filtrates were purified with FPLC affinity chromatography. 4) The elute from FPLC was used for T-lymphocyte proliferation and ELISA test. 5) Lastly, SDS-PAGE was used to determine the molecular weight and purity of the final product. RESULTS: A protein purified from pig spleen (the pig ISPS homologue) inhibited concanavalin A (Con A)-induced mouse lymphocyte proliferation. The molecular weight of this protein was about Mr 190 000. It has a stronger selectivity against T-lymphocyte line such as Jurkat cell line and mastocyte line (P8l5) and has a weaker inhibitory activity on macrophage line (U937). CONCLUSION: A protein similar to rat/mouse ISPS was found in pig spleen. This may provide an opportunity to study its roles in tumors and autoimmune diseases.

[A preliminary study on the immune suppressive protein of stress in human tonsil].

Our previous work demonstrated that under the conditions of restraint stress and under the control of central nervous system (CNS), an immune suppressive protein of stress (ISPS) was generated in peripheral lymph tissue and released into the blood stream, acting as an immune suppressor. In the present work, a protein similar to ISPS was found in human tonsil (a peripheral lymph tissue). Human tonsil was homogenized and the extract was prepared. It was found that lymphocyte proliferation was significantly suppressed by the extract. The suppression induced by the extract was partially reversed by the monoclonal antibody against ISPS (2C4). In ELISA test, the extract was able to bind to the monoclonal antibody. By immunohistochemistry, many ISPS positive cells were found in human tonsil. The ISPS positive cells were also found in human lymph nodes. Taken together, all the results demonstrate that a protein similar to ISPS may exist in human peripheral lymphoid tissue.

[Calcitonin gene-related peptide induces chemokine interleukine-8 synthesis in human monocytes].

OBJECTIVE: To study the effect of calcitonin gene-related peptide (CGRP) on the function of immune cells. METHODS: Human monocytes were cultured with calcitonin gene-related peptide in vitro and activated with lipopolysaccharide (LPS). The IL-8 level in the supernatant was measured with ELISA and the IL-8 mRNA expression in monocytes was observed by reverse transcription polymerase chain reaction (RT-PCR). The chemotactic activity of monocytes to neutrophils and lymphocytes was analyzed with micro-chemotacxis chamber. Chemotactic index (CI) was calculated by the formula: number of monocytes migrating to the underside of membrane in the LPS + CGRP group/number of monocytes migrating to the underside of membrane in the control group. CGRP receptor antagonist CGRP8 - 37 was added into the culture to study the effect of CGRP. Blank control and cultures of monocytes with LPS or with CGRP only were used as controls. RESULTS: The level of IL-8 protein in the supernatant of the LPS + CGRP group was 1 120 pg/ml +/- 14.80 pg/ml, significantly higher than those in other groups (670 pg/ml +/- 15.10 pg/ml in LPS + CGRP + CGRP8 - 37 group). The expression of IL-8 mRNA in the LPS + CGRP group was the highest (IL-8/beta-actin = 1.845 +/- 0.587), IL-8/beta-actin in the LPS + CGRP + CGRP8 - 37 group was1.339 +/- 0.434. The chemotactic activities of the monocytes to neutrophils and lymphocytes were enhanced in the LPS + CGRP group (CI = 3.78 +/- 0.08 to neutrophils and CI = 3.4 +/- 0.27 to lymphocytes). The CI values were 1.15 +/- 0.31 and 1.21 +/- 0.06 respectively in the LPS + CGRP + CGRP 8 - 37 group. CONCLUSION: CGRP in the peripheral nerve ending induces monocytes to synthetize and secret chemotactic factor IL-8 and enhance the chemotactic activity of monocytes, thus promoting the directional migration and aggregation of neutrophils and lymphocytes to foci of inflammation.