Network pharmacology combined with experimental validation to investigate the effect of Rongjin Niantong Fang on chondrocyte apoptosis in knee osteoarthritis.

Knee osteoarthritis (KOA) is a chronic degenerative disease that affects the quality of life of middle­aged and elderly individuals, and is one of the major factors leading to disability. Rongjin Niantong Fang (RJNTF) can alleviate the clinical symptoms of patients with KOA, but the molecular mechanism underlying its beneficial effects on KOA remains unknown. Using pharmacological analysis and in vitro experiments, the active components of RJNTF were analyzed to explore their potential therapeutic targets and mechanisms in KOA. The potential targets and core signaling pathways by which RJNTF exerts its effects on KOA were obtained from databases such as Gene Expression Omnibus, Traditional Chinese Medicine Systems Pharmacology and Analysis Platform. Subsequently, chondrocyte apoptosis was modeled using hydrogen peroxide (H2O2). Cell Counting Kit­8 assay involving a poly [ADP­ribose] polymerase­1 (PARP1) inhibitor, DAPI staining, reverse transcription­quantitative PCR, Annexin V­FITC/PI staining and flow cytometry, western blotting and co­immunoprecipitation analysis were used to determine the therapeutic efficacy of RJNTF on KOA and to uncover the molecular mechanism. It was found that PARP1­knockdown lentivirus, incubation with PARP1 inhibitor PJ34, medium and high doses of RJNTF significantly reduced H2O2­induced chondrocyte apoptosis. Medium and high doses of RJNTF downregulated the expression of cleaved caspase­3, cleaved PARP1 and PAR total proteins, as well as nucleus proteins of apoptosis­inducing factor (AIF) and migration inhibitory factor (MIF), and upregulated the expression of caspase­3, PARP1 total protein, as well as the cytoplasmic expression of AIF and MIF, suggesting that RJNTF may inhibit chondrocyte apoptosis through the PARP1/AIF signaling pathway.

Rongjin Niantong Fang ameliorates cartilage degeneration by regulating the SDF-1/CXCR4-p38MAPK signalling pathway.

CONTEXT: Rongjin Niantong Fang (RJNTF) is a Traditional Chinese Medicine formulation with a good therapeutic effect on osteoarthritis (OA). However, the underlying mechanisms remain unclear. OBJECTIVE: This study investigates whether RJNTF could delay OA cartilage degeneration by regulating the SDF-1/CXCR4-p38MAPK signalling pathway. MATERIALS AND METHODS: The Sprague-Dawley (SD) rats were used to establish the OA model by a modified Hulth's method. SD rats were divided into three groups (n = 10): blank group, model group (0.9% saline, 10 mL/kg/day), and treatment group (RJNTF, 4.5 g/kg/day). After 12 weeks of treatment, each group was analysed by H&E, Safranine-O solid green, ELISA, Immunohistochemistry, and Western blot. An in vitro model was induced with 100 ng/mL SDF-1 by ELISA, the blank group, model group, RJNTF group, and inhibitor group with intervention for 12 h, each group was analysed by Immunofluorescence staining and Western blot. RESULTS: SDF-1 content in the synovium was reduced in RJNTF treatment group compared to non-treatment model group (788.10 vs. 867.32 pg/mL) and down-regulation of CXCR4, MMP-3, MMP-9, MMP-13 protein expression, along with p38 protein phosphorylated were observed in RJNTF treatment group. In vitro results showed that RJNTF (IC50 = 8.925 mg/mL) intervention could down-regulate SDF-1 induced CXCR4 and p38 protein phosphorylated and reduce the synthesis of MMP-3, MMP-9, and MMP-13 proteins of chondrocytes from SD rat cartilage tissues. DISCUSSION AND CONCLUSION: RJNTF alleviates OA cartilage damage by SDF-1/CXCR4-p38MAPK signalling pathway inhibition. Our ongoing research focuses on Whether RJNTF treats OA through alternative pathways.

Proteome Analysis of the Soybean Nodule Phosphorus Response Mechanism and Characterization of Stress-Induced Ribosome Structural and Protein Expression Changes.

In agroecosystems, a plant-usable form of nitrogen is mainly generated by legume-based biological nitrogen fixation, a process that requires phosphorus (P) as an essential nutrient. To investigate the physiological mechanism whereby phosphorus influences soybean nodule nitrogen fixation, soybean root nodules were exposed to four phosphate levels: 1 mg/L (P stress), 11 mg/L (P stress), 31 mg/L (Normal P), and 61 mg/L (High P) then proteome analysis of nodules was conducted to identify phosphorus-associated proteome changes. We found that phosphorus stress-induced ribosomal protein structural changes were associated with altered key root nodule protein synthesis profiles. Importantly, up-regulated expression of peroxidase was observed as an important phosphorus stress-induced nitrogen fixation-associated adaptation that supported two nodule-associated activities: scavenging of reactive oxygen species (ROS) and cell wall growth. In addition, phosphorus transporter (PT) and purple acid phosphatase (PAPs) were up-regulated that regulated phosphorus transport and utilization to maintain phosphorus balance and nitrogen fixation function in phosphorus-stressed root nodules.

First Report of Fusarium Wilt of Industrial Hemp (Cannabis sativa L.) Caused by Fusarium oxysporum in the Northeast China.

Industrial hemp (Cannabis sativa L.) is an ancient and economically important crop used in food, medicine, textile, and paper industries (Chandra et al. 2017). In July 2021, an estimated 30% of the industrial hemp plants showed wilted leaves and root rot in the greenhouse at the Modern Agriculture Demonstration Area Management Center, Harbin City, Heilongjiang Province, China. Initially, the diseased plants exhibited green and reversible wilting of lower canopy leaves. Upon progression the plants showed irreversible wilting. The epidermal tissue of root and rhizome showed slight cracks and the vascular bundle exhibited light brown discoloration, and then died. Six randomly selected disease plants were collected. Small fragments (5 mm) were cut from the infected roots, surface-sterilized with 70% ethanol for 30s and 1% sodium hypochlorite for 5 min and rinsed three times in sterile H2O. Then the small pieces were embedded on potato dextrose agar at 25℃ for 4 days and sub-cultured by hyphal tipping to isolate the fungus. A single-spore culture was obtained by monosporic isolation. The colonies were characterized by an abundant white cottony mycelium, which became gray or purple with age. The macroconidia were transparent, short to medium in length, straight to slightly curved, septate 0 to 4, 16.8 to 26.6 µm long × 3.5 to 4.1 µm wide. The apical cells were long and tapering to a point and the basal cells were notched. Microconidia were elliptic or kidney-shaped, and septate 0 to 4. The conidia were 4.2 to 11.3 µm long × 3.5 to 5.5 µm wide (n = 50). The morphological characteristics were very similar to those of Fusarium oxysporum (Leslie and Summerell 2006). For molecular identification, the internal transcribed spacer (ITS), translation elongation factor 1-α (TEF1) and RNA polymerase II beta subunit (RPB2) genes were amplified and sequenced with the primers ITS1/ITS4, EF-1/EF-2 (Uwaremwe et al. 2020), and 5f2/7c (O'Donnell et al. 2010). The 520 bp (ITS), 948 bp (TEF1), and 861 bp (RPB2) sequences were deposited in GenBank with acce. nos. MZ722998, OK180473 and OK180474, respectively. NCBI BLAST analysis showed 98 to 100% similarity with the sequences of F. oxysporum. Moreover, the sequences alignment similarity for the six isolates were 100%. Based on the morphological and molecular characteristics, the isolates were identified as F. oxysporum. For the pathogenicity test, 20 seedlings were inoculated 30 ml of a conidial suspension (106 conidia/ml) using the root dip method. Another set of 20 seedlings were inoculated with the same quantity of sterile distilled water as the controls. After inoculation, all seedlings were maintained in a greenhouse at 25°C ± 2, with a relative humidity of 60 to 70% and a 16 h light/8 h dark cycle. This test was repeated twice. The leaves of the inoculated seedlings gradually became yellow and exhibited wilting within 15 to 20 days, the epidermal tissue of root showed light brown discoloration. Eventually the plants were dead within 40 to 50 days after inoculation. The control seedlings did not show any wilt symptoms. F. oxysporum was re-isolated from the infected root tissues to fulfill the Koch's postulates. In addition to F. oxysporum, F. brachygibbosum, Pythium aphanidermatum, F. solani, and F. equiseti have also been reported to cause wilt symptoms of industrial hemp (Zamir et al. 2018). To our knowledge, this is the first report of Fusarium wilt on C. sativa caused by F. oxysporum in the Northeast China.

An electrochemiluminescence biosensor for cadmium ion based on target-induced strand displacement amplification and magnetic Fe3O4-GO nanosheets.

Taking advantage of an exquisite hairpin DNA for strand displacement amplification (SDA) and the magnetic Fe3O4-graphene oxide nanosheets (MGN) as the carrier, an immobilization-free ECL biosensor was constructed for ultra-trace detection of Cd2+. Firstly, the ECL probe Ru (phen)32+ easily diffuses in the solution and reaches the electrode surface to induce strong ECL signal. This is because the pre-designed hairpin DNA is constrained by MGN in the absence of Cd2+. The presence of Cd2+ releases cDNA by binding to its corresponding aptamer, leading to removal of hairpin DNA away from the surface of MGN. In this case, SDA amplification was evoked and generated numerous dsDNA which further trapped Ru (phen)32+ in its groove. It is difficult for the embedded ECL probe to touch the electrode surface to generate ECL signal. Therefore, the concentration of Cd2+ was monitored according to the attenuation of ECL signal. This method showed high sensitivity to Cd2+ with a detection limit of 1.1 × 10-4 ppb. Moreover, it not only avoids many condition optimizations required in the conventional SDA method, but also circumvent the modification and immobilization of DNA probe. This sensor is further applied in the detection of Cd2+ in the sample of traditional Chinese medicine.

Xinshuitong Capsule extract attenuates doxorubicin-induced myocardial edema via regulation of cardiac aquaporins in the chronic heart failure rats.

Doxorubicin (Dox), an effective antineoplastic drug, was limited use for cardiotoxicity. Xinshuitong Capsule (XST), a patented herbal formula, showed desirable beneficial effects in the treatment of chronic heart failure (CHF) patients. However, the drug on Dox-induced cardiotoxicity remains unclear. Ninety male Sprague-Dawley rats were randomized into two groups: 15 rats were selected as the normal group and 75 rats were injected intraperitoneally with Dox to establish CHF rat models, the success ones were randomly divided into five groups: low XST (LXST), medium XST (MXST) or high XST (HXST) (4.9, 9.8, or 19.6 g/kg d) administrated intragastrically twice a day for 4 weeks, with the captopril-treated group and the model group as comparison. The model group showed the cardiac functions generally impaired, and CHF mortality rate higher (47%) than those in the XST-treated groups (averaged 24%, P < 0.05). Compared with XST-treated groups, myocardial remodeling, inflammation and desarcomerization, and higher water content more severe in the cardiac tissue in the model group (P < 0.05), which was associated with higher expressions of mRNA or protein levels of AQP1, 4 and 7. Dox-impaired cardiac functions, cardiac remodeling and myocardial edema could be dose-dependently reverted by XST treatment. XST could inhibit AQP1, 4 and 7 at mRNA levels or at protein levels, which was associated with the attenuation of myocardial edema and cardiac remodeling, decreasing the ventricular stiffness and improving the cardiac functions and rats' survival. AQPs is involved in cardiac edema composed one of the mechanisms of Dox-induced cardiotoxicity, XSTvia inhibition of AQPs relieved the Dox-induced side effects.

Genome-wide identification and expression analysis of the WRKY transcription factor family in flax (Linum usitatissimum L.).

BACKGROUND: Members of the WRKY protein family, one of the largest transcription factor families in plants, are involved in plant growth and development, signal transduction, senescence, and stress resistance. However, little information is available about WRKY transcription factors in flax (Linum usitatissimum L.). RESULTS: In this study, comprehensive genome-wide characterization of the flax WRKY gene family was conducted that led to prediction of 102 LuWRKY genes. Based on bioinformatics-based predictions of structural and phylogenetic features of encoded LuWRKY proteins, 95 LuWRKYs were classified into three main groups (Group I, II, and III); Group II LuWRKYs were further assigned to five subgroups (IIa-e), while seven unique LuWRKYs (LuWRKYs 96-102) could not be assigned to any group. Most LuWRKY proteins within a given subgroup shared similar motif compositions, while a high degree of motif composition variability was apparent between subgroups. Using RNA-seq data, expression patterns of the 102 predicted LuWRKY genes were also investigated. Expression profiling data demonstrated that most genes associated with cellulose, hemicellulose, or lignin content were predominantly expressed in stems, roots, and less in leaves. However, most genes associated with stress responses were predominantly expressed in leaves and exhibited distinctly higher expression levels in developmental stages 1 and 8 than during other stages. CONCLUSIONS: Ultimately, the present study provides a comprehensive analysis of predicted flax WRKY family genes to guide future investigations to reveal functions of LuWRKY proteins during plant growth, development, and stress responses.

Anti-inflammatory activity of Radix Angelicae biseratae in the treatment of osteoarthritis determined by systematic pharmacology and in vitro experiments.

Radix Angelicae biseratae is a widely used Chinese traditional herbal medicine for osteoarthritis (OA). Its therapeutic efficacy has been confirmed in clinical practice. However, its mechanisms of action in treating OA have remained elusive. The purpose of the present study was to identify active components with good oral bioavailability and drug-like properties from Radix Angelicae biseratae through systematic pharmacology and in vitro experiments to determine targets of Radix Angelicae biseratae in the treatment of OA. The functional components of Radix Angelicae biseratae were screened from the Traditional Chinese Medicine Systems Pharmacology database based on oral bioavailability and drug-like properties. Subsequently, the databases STITCH, Open Targets Platform and DrugBank were searched and microarray analysis was performed to screen the active components of Radix Angelicae biseratae to treat OA and predict its potential target proteins. The interaction network and protein interaction network were then generated and examined, molecular docking between active components and targets was performed and the enrichment of potential target proteins was analyzed. Finally, reverse transcription-quantitative (RT-q)PCR and western blot analyses were used to verify the therapeutic effect of Radix Angelicae biseratae extract on the expression of OA-associated target proteins. The results provided eight active components in Radix Angelicae biseratae, which were firmly linked to 20 targets of OA. In combination with molecular docking and the analysis of the interaction network between components and targets, it was suggested that sitosterol was a major active component of Radix Angelicae biseratae in the treatment of OA. Protein interaction network analysis suggested that prostaglandin-endoperoxide synthase 2 (PTGS2), nitric oxide synthase 3 and cytochrome P450 2B6 may be critical targets for Radix Angelicae biseratae in the treatment of OA. In addition, RT-qPCR and western blot analyses suggested that Radix Angelicae biseratae extract inhibited the mRNA and protein expression of PTGS2 in degenerative articular cartilage cells in vitro, whilst other targets remain to be verified. Functional enrichment analysis indicated that Radix Angelicae biseratae confers pharmacological efficacy towards OA through exerting anti-inflammatory effects and immune regulation.

A Systematic Pharmacology and In Vitro Study to Identify the Role of the Active Compounds of Achyranthes bidentata in the Treatment of Osteoarthritis.

BACKGROUND Achyranthes bidentata is a Chinese traditional herbal medicine widely used to treat osteoarthritis (OA). This study aimed to identify active compounds from Achyranthes bidentata through systematic pharmacology and in vitro experiments to find the targets of Achyranthes bidentata in the treatment of OA. MATERIAL AND METHODS We screened the active compounds of Achyranthes bidentata from the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Then, we used STITCH and Open Targets Platform databases to screen the active components and predict the potential targets of Achyranthes bidentata in the treatment of OA. Subsequently, we studied the compound-target network and protein interaction network and analyzed the enrichment of potential target proteins. Finally, we used Western blot analysis to verify the therapeutic effect of Achyranthes bidentata extract on the expression of OA-related target proteins. RESULTS There were 7 active components in Achyranthes bidentata, which were strongly related to the 74 targets of OA. Quercetin, baicalein, and berberine are the critical active compounds of Achyranthes bidentata in the treatment of OA. Protein interaction analysis and in vitro experiments suggested that TNF, IL-6, and TP53 are the critical targets of Achyranthes bidentata in the treatment of OA. Functional enrichment analysis showed that Achyranthes bidentata plays a pharmacological role in OA through apoptosis, inflammation, and immune regulation. CONCLUSIONS Quercetin, baicalein, and berberine are the critical active compounds of Achyranthes bidentata in the treatment of OA. TNF, IL-6, and TP53 may be potential targets for the treatment of OA.

Tougu Xiaotong capsule exerts a therapeutic effect by improving knee meniscus in the early osteoarthritis rat model.

The aim of the study was to observe the effects of Tougu Xiaotong capsule (TGXTC) on the microstructure and ultrastructure of meniscus in rats with early knee osteoarthritis (KOA). A total of 27 Sprague Dawley rats were randomly divided into three groups: The normal group (non-papain-induced KOA; received saline only), the model group (papain-induced KOA; received saline only) and the TGXTC group [papain-induced KOA; received TGXTC (0.31g·kg-1·d-1)]. After 4 weeks treatment, the animals were anesthetized and the sagittal plane of the intact knees (n=6 per group) was obtained and prepared in paraffin section. Following hematoxylin and eosin staining, the degeneration of cartilage structure was evaluated via Mankin score, the microstructure of meniscus was observed and the area of calcification in meniscus was analyzed. Following toluidine blue staining, the content of proteoglycan in meniscus was analyzed. Three samples in each group were obtained and the ultrathin sections of meniscus were observed through a transmission electron microscope. The results showed that compared with the normal group, in the model group the joint space became narrow and the cartilage layer was slightly damaged and the Mankin score was 4.17±0.76, suggesting that the early KOA model was successfully established. After TGXTC treatment, the joint space stenosis and cartilage damage were improved as the Mankin score significantly decreased. Compared with the normal group, in the model group the surface of meniscal cartilage was much more uneven, the area of calcification was significantly increased and the content of proteoglycan of cartilage matrix was significantly decreased. However, following TGXTC treatment, the surface of the meniscal cartilage was much more smooth and flat, and the damage of tissue structure and the calcified area were significantly reduced, and the proteoglycan of cartilage matrix content was significantly increased. Compared with the normal group, the number of cellular processes and organelles, including the rough endoplasmic reticulum, mitochondria and Golgi apparatus of meniscal cartilage were reduced and swollen in the model group. In addition, the nuclei were deformed and heterochromatin agglutinated. The extracellular collagen fibrils became slender, disordered and sparse. Compared with the model group, the TGXTC group had more cell processes and organelles, alleviated swelling and heterochromatin agglutinating. Additionally, the collagen fibrils around the cells were thicker, larger and arranged in an orderly manner. In conclusion, TGXTC exerted its therapeutic effects on the development of KOA via reducing the destruction of the cartilage structure of the meniscus and improving the composition and function of the meniscus cartilage matrix.

Development of a specific AFLP-based SCAR marker for Chinese Race 34MKG of Puccinia graminis f. sp. tritici.

Wheat stem rust, caused by Puccinia graminis f. sp. tritici (Pgt), is a fungus that causes the devastating fungalwheat stem rust disease in wheat production. Rapid identification of the physiological races of Pgt are very importance for the prevention of wheat stem rust. In this paper we developed a molecular method to identify the most prevalent race of Pgt, as a supplement for traditionally used host-specific methods. Amplified fragment length polymorphism (AFLP) was employed as a means of analyzing DNA polymorphisms in six common physiological races of Pgt in China and Ug99. In total, 64 pairs of primers were used for AFLP screening of race-specific molecular markers. One primer pair-namely, E7/M7 (5'-GACTGCGTACCAATTCG G-3'/5'-GATGAGTCCTGAGTAACGG-3')-yielded a unique band for the race 34MKG that was purified and cloned into the pGEM-T vector for sequencing. We then designed a new primer pairs (sequence-characterized amplified region marker) to amplify the 171-bp fragment and confirmed that the marker was highly specific for 34MKG. These results provide a new tool for monitoring different races of Pgt for improved control of wheat stem rust in China.

[Retracted] Differential miRNAomics of the synovial membrane in knee osteoarthritis induced by bilateral anterior cruciate ligament transection in rats.

Following the publication of the above article, a number of errors were identified in the paper, and after having consulted with the editor of Molecular Medicine Reports, a corrigendum was published last year ("[Corrigendum] Differential miRNAomics of the synovial membrane in knee osteoarthritis induced by bilateral anterior cruciate ligament transection in rats." Zhou J, Zhao Y, Wu G, Lin B, Li Z and Liu X. Mol Med Rep 20: 5363, 2019). However, following publication of the above corrigendum, the paper was re­examined by the authors, and additional errors were identified; therefore, the authors are going to retract this paper from the publication. All the authors agree to this retraction, and apologize to the Editor of Molecular Medicine Reports and to the readership for any inconvenience caused. [the original article was published in Molecular Medicine Reports 18: 4051­4057, 2018; DOI: 10.3892/mmr.2018.9385].

[Corrigendum] Differential miRNAomics of the synovial membrane in knee osteoarthritis induced by bilateral anterior cruciate ligament transection in rats.

Following the publication of the above article, an interested reader drew to our attention that, in Fig. 4A, in which the authors had presented a western blot image depicting protein expression of IL­18, IL­1ß, NLRP3 and ß­actin from synovial tissue lysates from osteoarthritic rats, upon close examination of the figure a striking similarity was noted between the bands shown for the IL­18­ and NLRP3­stained Sham group experiments, although the bands appeared in an inverted position relative to each other. Following an enquiry with the authors, they realized that they had included incorrect data for this figure; an amended version of Fig. 4, showing the correct data for NLRP3, is shown opposite. Secondly, the authors have realized that, at various points throughout the paper, two miRNAs were written incorrectly: Specifically, references to an 'miR­352' should have appeared as miR­532, and 'miR­233' should have been written as miR­223. This error affected the presentation of Fig. 2; therefore, a revised version of this figure, with the miRNAs correctly labelled as miR­532 and miR­223 respectively, is also shown opposite. Furthermore, mi­233 should have been written as miR­223 at the following places in the text: p. 4051, right­hand column (RHC), line 10 ("Furthermore, the miR­223­regulated...); p. 4054, Results section, left­hand column (LHC), third subheading ("miR­223 negatively regulates the expression of NLRP3."); and in the concluding paragraph of the Discussion on p. 4056, LHC, miR­233 should have been written as miR­223 in all five instances where this occurred (lines 4, 7, 8, 9, and 10 of this paragraph). Finally, the second sentence featured in the subsection of the Results section entitled "Expression validation of miRs by RT­qPCR" on p. 4054 contained additional errors. This sentence should have appeared as follows (changed text is highlighted in bold): "The expression of miR­223, ­100, ­345, ­130, ­382, ­9a and 183 were upregulated, whereas miR­377, ­532, ­200b were downregulated with a fold change of ≥1.5, similar to the microarray data (Fig. 2). All the authors agree to the contents of this Corrigendum, and apologize to the Editor of Molecular Medicine Reports and to the readership for any inconvenience caused. [the original article was published in Molecular Medicine Reports 18: 4051­4057, 2018; DOI: 10.3892/mmr.2018.9385].

Inflammatory cytokines via up-regulation of aquaporins deteriorated the pathogenesis of early osteoarthritis.

BACKGROUND: Inflammatory cytokines enhanced the progress of the pathogenesis of osteoarthritis, however the mechanisms remain unclear. The objective is to determine aquaporins (AQPs) in the pathogenesis of osteoarthritis. METHODS AND FINDINGS: Primary rat articular chondrocytes were treated with IL-1ß to mimic the early stage of osteoarthritis in vitro. Early osteoarthritis animal model was established by intra-articular injection of 4% papain. Micro- or ultra-structure histopathologic changes, cell viability, apoptosis cells and cell membrane permeability, locations and expressions of AQP1 and AQP3 and matrix were detected in the cartilage or in the chondrocytes of knee. IL-1ß could reduce the chondrocytes viability, increase the apoptosis cells, and also impair the cell membrane and organelles. IL-1ß significantly induced the up-regulation of AQP1 and AQP3 in the chondrocytes. In the chondrocytes, AQPs were mainly clustered in both membrane and perinuclear region of cytoplasm, while higher AQPs were detected in the superficial and middle layers of the cartilage. With the up-regulation of AQPs, the cartilage matrix was considerably decreased in both the chondrocytes and in the osteoarthritis cartilage. In the early osteoarthritis rat model, serum and synovial fluid confirmed that higher IL-1ß could increase the expressions of AQPs, and decrease the cartilage matrix in both the chondrocytes and the cartilage. CONCLUSIONS: Inflammatory cytokine IL-1ß via up-regulation of AQPs caused the abnormal metabolism of water transport and loss of the cartilage matrix in the chondrocytes, and ultimately exacerbated the pathogenesis of early osteoarthritis. Therefore, AQPs may be a candidate therapeutic target for prevention and treatment of osteoarthritis.

Management of Combined Atlas Fracture with Type II Odontoid Fracture: A Review of 21 Cases.

PURPOSE: To evaluate the therapeutic effects of combined atlas fracture with type II (C1-type II) odontoid fractures and to outline a management strategy for it. PATIENTS AND METHODS: Twenty three patients with C1-type II odontoid fractures were treated according to our management strategy. Nonoperative external immobilization in the form of cervical collar and halo vest was used in 13 patients with stable atlantoaxial joint. Surgical treatment was early performed in 10 patients whose fractures with traumatic transverse atlantal ligament disruption or atlantoaxial instability. The visual analog scale (VAS), neck disability index (NDI) scale, and American Spinal Injury Association (ASIA) scale at each stage of followup were then collected and compared. RESULTS: Compared to pretreatment, the VAS score, NDI score, and ASIA scale were improved among both groups at followup evaluation after treatment. However, in the nonsurgical group, one patient (1/11) developed nonunion which required surgical treatment in later stage and one patient (1/13) with halo vest immobilization had happened pin site infection. Two patients of the surgical group (2/11) had appeared minor complications: occipital cervical pain in one case and cerebrospinal fluid leakage in one case. Two patients (2/23) were excluded from nonsurgical treatment group because their followup period was less than 12 months. Twenty one patients were followed up regularly with an average of 23.9 months (range 15-45 months). CONCLUSIONS: We outlined our concluding management principle for the treatment of C1-type II odontoid fractures based on the nature of C1 fracture and atlantoaxial stability. The treatment principle can obtain satisfactory results for the management of C1-type II odontoid fractures.

[Acupuncture combined with cinesiotherapy cupping for knee osteoarthritis with qi stagnation and blood stasis syndrome: a randomized controlled trial].

OBJECTIVE: To compare the clinical efficacy between acupuncture combined with cinesiotherapy cupping and acupuncture combined with conventional cupping for knee osteoarthritis (KOA) with qi stagnation and blood stasis syndrome, and to seek a better solution for KOA. METHODS: A total of 78 patients of KOA with qi stagnation and blood stasis syndrome were randomly divided into an observation group and a control group, 39 cases in each group (3 cases in the observation group and 2 cases in the control group lost contact). Both groups were treated with acupuncture at Neixiyan (EX-LE 4), Dubi (ST 35), Xuehai (SP 10), Liangqiu (ST 34), Heding (EX-LE 2), Zusanli (ST 36), Yinlingquan (SP 9), Yanglingquan (GB 34) and Xuanzhong (GB 39). Based on the acupuncture treatment, the control group was treated with conventional cupping. The No. 4 cupping glass was used for Xuehai (SP 10), Liangqiu (ST 34) and Fengshi (GB 31), while the No. 3 cupping glass was used for Yinlingquan (SP 9), while the cupping with appropriate size was used for ashi points; the cupping glass was retained for 5 min. Based on the acupuncture treatment, the observation group was treated with cinesiotherapy cupping. The selection of acupoint and cupping glass was identical as the control group. The patients were instructed to perform knee flexion-extension, hip abduction-adduction, weight-bearing and other active exercise while cupping; the treatment was given once a day, 10 times as a course of treatment; totally three courses were given with an interval of 2 days between the courses. The patient's symptom scores, pain scores and knee function scores were recorded before and after treatment. The amount of joint effusion was measured by ultrasound; the level of interleukin-1 (IL-1), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) in joint effusion were measured by ELISA. RESULTS: After treatment, the total effective rate in the observation group was 94.4% (34/36), which was significantly higher than 86.5% (32/37) in the control group (P<0.05). Compared before treatment, the symptom scores, pain scores, amount of joint effusion and the levels of IL-1, IL-6, TNF-α in joint fluid in both groups all were decreased after treatment, whereas the knee function scores were increased (P<0.05). Compared with the control group, the symptom scores, pain scores, and the levels of IL-1, TNF-α and the amount of joint effusion all were significantly decreased, whereas the knee function scores were increased in the observation group (P<0.05). The level of IL-6 in joint effusion was not significantly different between the observation group and the control group (P>0.05). CONCLUSION: The acupuncture combined with cinesiotherapy cupping could alleviate pain, improve joint function and reduce joint effusion, which is superior to acupuncture combined with conventional cupping.

Exogenous Application of Phytohormones Promotes Growth and Regulates Expression of Wood Formation-Related Genes in Populus simonii × P. nigra.

Although phytohormones are known to be important signal molecules involved in wood formation, their roles are still largely unclear. Here, Populus simonii × P. nigra seedlings were treated with different concentrations of exogenous phytohormones, indole-3-acetic acid (IAA), gibberellin (GA3), and brassinosteroid (BR), and the effects of phytohormones on growth were investigated. Next, 27 genes with known roles in wood formation were selected for qPCR analysis to determine tissue-specificity and timing of responses to phytohormone treatments. Compared to the control, most IAA, GA3, and BR concentrations significantly increased seedling height. Meanwhile, IAA induced significant seedling stem diameter and cellulose content increases that peaked at 3 and 30 mg·L-1, respectively. Significant increase in cellulose content was also observed in seedlings treated with 100 mg·L-1 GA3. Neither stem diameter nor cellulose content of seedlings were affected by BR treatment significantly, although slight effects were observed. Anatomical measurements demonstrated improved xylem, but not phloem, development in IAA- and BR-treated seedlings. Most gene expression patterns induced by IAA, GA3, and BR differed among tissues. Many IAA response genes were also regulated by GA3, while BR-induced transcription was weaker and slower in Populus than for IAA and GA3. These results reveal the roles played by phytohormones in plant growth and lay the foundation for exploring molecular regulatory mechanisms of wood formation in Populus.

Electroacupuncture Delays Cartilage Degeneration by Modulating Nuclear Factor-κB Signaling Pathway.

OBJECTIVE: To illustrate the molecular mechanisms underlying the therapeutic effects of electroacupuncture (EA) on knee osteoarthritis (OA). METHODS: Twenty-seven six-month-old New Zealand white rabbits were allocated into three groups in accordance with a random number table: normal group (no surgery-induced OA; without treatment), model group (surgery-induced OA; without treatment) and EA group [surgery-induced OA; received treatment with EA at acupoints Dubi (ST 35) and Neixiyan (EX-LE 5), 30 min twice a day]. After eight consecutive weeks of treatment, the histopathological alterations in cartilage were observed using optical microscopy and transmission electron microscopy, cartilage degeneration was evaluated by modified Mankin's score principles, the synovial fluid concentration of interleukin-1ß (IL-1ß), interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α) and matrix metalloproteinase-3 (MMP-3) were evaluated by enzyme-linked immunosorbent assay, and the protein expression levels of IL-1ß, IL-6, TNF-α, MMP-3, IκB kinase-ß (IKK-ß), nuclear factor of α light polypeptide gene enhancer in B-cells inhibitor α (IκB-α) and nuclear factor-κB (NF-κB) p65 were quantified by Western blot analysis. RESULTS: EA treatment significantly improved cartilage structure arrangement and reduced cellular degeneration. The IL-1ß, IL-6, TNF-α and MMP-3 of synovial fluid in the EA-treated group were significantly decreased compared with the model group (all P<0.01). Compared with the model group, the IL-1ß, IL-6, TNF-α, MMP-3, IKK-ß and NF-κB p65 protein expressions in cartilage of EA-treated group were significantly decreased (all P<0.01), whereas IκB-α expression was significantly up-regulated (P<0.01). CONCLUSION: EA treatment may delay cartilage degeneration by down-regulating inflammatory factors through NF-κB signaling pathway, which may, in part, explain its clinical efficacy in the treatment of knee OA.

Comprehensive Analysis of Differentially Expressed Unigenes under NaCl Stress in Flax (Linum usitatissimum L.) Using RNA-Seq.

Flax (Linum usitatissimum L.) is an important industrial crop that is often cultivated on marginal lands, where salt stress negatively affects yield and quality. High-throughput RNA sequencing (RNA-seq) using the powerful Illumina platform was employed for transcript analysis and gene discovery to reveal flax response mechanisms to salt stress. After cDNA libraries were constructed from flax exposed to water (negative control) or salt (100 mM NaCl) for 12 h, 24 h or 48 h, transcription expression profiles and cDNA sequences representing expressed mRNA were obtained. A total of 431,808,502 clean reads were assembled to form 75,961 unigenes. After ruling out short-length and low-quality sequences, 33,774 differentially expressed unigenes (DEUs) were identified between salt-stressed and unstressed control (C) flax. Of these DEUs, 3669, 8882 and 21,223 unigenes were obtained from flax exposed to salt for 12 h (N1), 24 h (N2) and 48 h (N4), respectively. Gene function classification and pathway assignments of 2842 DEUs were obtained by comparing unigene sequences to information within public data repositories. qRT-PCR of selected DEUs was used to validate flax cDNA libraries generated for various durations of salt exposure. Based on transcriptome sequences, 1777 EST-SSRs were identified of which trinucleotide and dinucleotide repeat microsatellite motifs were most abundant. The flax DEUs and EST-SSRs identified here will serve as a powerful resource to better understand flax response mechanisms to salt exposure for development of more salt-tolerant varieties of flax.

Tougu Xiaotong capsule exerts a therapeutic effect on knee osteoarthritis by regulating subchondral bone remodeling.

Previous studies have shown that Tougu Xiaotong capsule (TGXTC) has therapeutic effects on knee osteoarthritis (OA) through multiple targets. However, the mechanisms of action underlying its regulation of subchondral bone reconstruction remain unclear. In this study, we investigated the effects of TGXTC on subchondral bone remodeling. Eighteen six-month-old New Zealand white rabbits of average sex were randomly divided into the normal, model and TGXTC groups. The rabbit knee OA model was induced by a modified Hulth's method in the model and TGXTC groups, but not the normal group. Five weeks postoperatively, intragastric administration of TGXTC was performed for four weeks. After drug administration, the medial femoral condyle and tibia were prepared for observation of cartilage histology via optical microscopy and micro-computed tomography, the serum was collected for biochemical parameters assay and the subchondral bone isolated from the lateral femoral condyle was collected for detection of IL-1ß and TNF-α mRNA and protein by reverse transcription-quantitative polymerase chain reaction and western blot analysis, respectively. The results showed that treatment with TGXTC significantly mitigated cartilage injury and subchondral bone damage, improved the parameter of subchondral trabecular bone, decreased alkaline phosphatase and tartrate-resistant acid phosphatase activity, and significantly reducing the osteoprotegerin/receptor activator of nuclear factor-κB ligand ratio, reduced the expression of IL-1ß and TNF-α mRNA and protein. These results suggest that TGXTC could delay the pathological development of OA by regulating subchondral bone remodeling through regulation of bone formation and bone resorption and its relating inflammatory factors, and this may partly explain its clinical efficacy in the treatment of knee OA.