PLCL1 regulates fibroblast-like synoviocytes inflammation via NLRP3 inflammasomes in rheumatoid arthritis

Abstract Background: Phospholipase C-like 1 (PLCL1), a protein that lacks catalytic activity, has similar structures to the PLC family. The aim of this research was to find the function and underlying mechanisms of PLCL1 in fibroblast-like synoviocyte (FLS) of rheumatoid arthritis (RA). Methods: In this study, we first analyzed the expression of PLCL1 in the synovial tissue of RA patients and K/BxN mice by immunohistochemical staining. Then silencing or overexpressing PLCL1 in FLS before stimulating by TNF-α. The levels of IL-6, IL-1β and CXCL8 in FLS and supernatants were detected by Western Blot (WB), Real-Time Quantitative PCR and Enzyme Linked Immunosorbent Assay. We used INF39 to specifically inhibit the activation of NLRP3 inflammasomes, and detected the expression of NLRP3, Cleaved Caspase-1, IL-6 and IL-1β in FLS by WB. Result: When PLCL1 was silenced, the level of IL-6, IL-1β and CXCL8 were down-regulated. When PLCL1 was overexpressed, the level of IL-6, IL-1β and CXCL8 were unregulated. The previous results demonstrated that the mechanism of PLCL1 regulating inflammation in FLS was related to NLRP3 inflammasomes. INF39 could counteract the release of inflammatory cytokines caused by overexpression of PLCL1. Conclusion: Result showed that the function of PLCL1 in RA FLS might be related to the NLRP3 inflammasomes. We finally confirmed our hypothesis with the NLRP3 inhibitor INF39. Our results suggested that PLCL1 might promote the inflammatory response of RA FLS by regulating the NLRP3 inflammasomes.

Sensitive and Rapid Detection of Traditional Chinese Herbs by Loop-Mediated Isothermal Amplification Method and Real-Time Fluorescence Quantitative PCR

ABSTRACT Adulterant herbal materials are threats to import and export trade and consumer safety. In this study, we established a simple and rapid examination system for the detection of Phellodendron chinense Schneid. Two detection methods, real-time fluorescence quantitative PCR (real-time PCR) and loop-mediated isothermal amplification (LAMP), were developed for traditional Chinese medicine detection, and their specificity and sensitivity were compared. The DNA of P. chinense was extracted and its special periods amplified with designed primers. Real-time PCR and LAMP experiments were conducted to test the specificity of primers in contrast to other similar species. The template concentration was diluted from 101 ng/µL to 10-5 ng/µL in order to contrast sensitivity between real-time PCR and LAMP. Real-time PCR and Lamp method has shown specificity because P. chinense was positive as opposed to other negative similar species. The Lamp method could detect a limited DNA concentration of 10-4ng/µL in 60 minutes with same sensitivity to real-time PCR. The results indicate that real-time PCR and LAMP are sensitive, accurate and specific in detection of P. chinense. However, LAMP is more convenient and cast less time. What's more, expensive equipments are not necessary for LAMP detector. For a better detection, we suggest an establishment of a real-time PCR and LAMP method for TCM market supervision which depends on DNA barcode sequences and LAMP.

Effect of rotavirus infection on regulating the expression of Na+-H+ exchanger 3 in Caco-2 cells via clathrin dependent endocytosis pathway / 中国感染与化疗杂志

Objective To observe the effect of rotavirus (RV) infection on expression level and bioactivity of Na+-H+ exchanger 3 (NHE3) in Caco-2 cells. Methods Model of NHE3-expressing Caco-2 cells was constructed and studied in terms of intervention: control, RV, clathrin antagonist chlorpromazine (CPZ), and CPZ + RV. NHE3 activity and NHE3 protein amount on cell surface were determined by BCECF-AM and biotinylation, respectively. Expression level of clathrin was assayed by Western blot. Results Compared with control group, NHE3 activity and NHE3 surface protein level significantly decreased when the cells were treated with RV. These effects could not be completely cancelled by clathrin antagonist CPZ. Moreover, RV treatment could increase cellular protein level of clathrin, which was cancelled by CPZ. Conclusions The effect of RV infection on NHE3 expression level and biological activity may be related to clathrin-dependent endocytosis pathway, and may be also affected by other endocytosis pathways.

Hypoxia Downregulates the Angiogenesis in Human Placenta via Notch1Signaling Pathway / 华中科技大学学报（医学）（英德文版）

Placentation,which is critical for maternal-fetal exchange of nutrients and gases,is a complicated process comprising stepwise vasculogenesis and angiogenesis.Hypoxia caused by impairedtrophoblast invasion may cause various angiogenic abnormalities in human placenta.The Notchl signaling pathway plays an important role in the regulation of angiogenesis.The angiogenesis of human umbilical vein endothelial cells (HUVECs) under normal/hypoxic conditions and the mRNA/protein level of Notchl/Dell4/Jaggedl were investigated in this study.The effects of DAPT/JAG-1 on the migration of HUVECs were also assessed by cell wound healing assay,so as to discover the possible role of notchl signaling pathway in the angiogenesis of human placenta.The results showed that angiogenic ability of HUVECs was seriously reduced under hypoxic conditions.The mRNA and protein levels of Notchl/Dell4/Jaggedl were decreased in the hypoxic group compared to the control one.In addition,the migration capability of HUVECs was significantly obstructed when treated with DAPT and under hopoxic condition,but promoted when treated with JAG-1.The above results demonstrate that hypoxia downregulates the angiogenesis in human placenta via Notch 1 signaling pathway.

Correlation of Stress Hyperglycemia after Ischemic Stroke with Early Vascular Cognitive Impairment / 中国医学科学院学报

Objective To investigate the influence and forecast value of stress hyperglycemia on the early vascular cognitive impairment (VCI) in stroke patients.Methods Totally 422 patients with acute non-diabetic stroke were divided into three groups according to the fasting plasma glucose level:the euglycemia group (<6.1 mmol/L),the mild stress hyperglycemia group (6.1-7.0 mmol/L),and the severe stress hyperglycemia group (≥7.0 mmol/L).Mini-mental state examination,Alzheimer's disease rating scale cognitive subscale,and clinical dementia rating scale were used to evaluate early cognition in post-stroke patients,and patients were divided into three groups accordingly:normal cognitive function group,mild VCI group,and vascular dementia group.Correlation analysis was carried out on the level of stress hyperglycemia and the degree of VCI.Results Of these 422 patients,stress hyperglycemia was identified in 62 cases (14.7%).The risk of stress hyperglycemia was higher in patients with a high degree of education [(8.39±3.85)years vs.(6.62±4.39)years,P=0.037)] or a history of cardiovascular disease (45.2% vs.18.3%,P=0.001).VCI was detected in 270 patients (64.0%).Age,sex,smoking,National Institute of Health Stroke Scale score,Hamilton Depression Rating Scale score,stress hyperglycemia,and history of cardiovascular disease were related with early VCI after non-diabetic ischemic stroke (P<0.05).Multivariate Logistic regression analysis showed that stress hyperglycemia was an independent risk factor for VCI in patients with non-diabetic ischemic stroke (OR=3.086,95% CI=1.065-8.929).The risks of cognitive impairment in the mild stress hyperglycemia group and the severe stress hyperglycemia group were higher than that of the euglycemia group,while it was also higher in the severe stress hyperglycemia group than in the mild stress hyperglycemia group (61.11% vs.75.00% vs.90.91%).Stress hyperglycemia was positively correlated with the high risk of early cognitive impairment in stroke patients (r=0.185,P=0.007).Conclusion There is a significant correlation between stress hyperglycemia and early VCI after ischemic stroke.

Impact of libido at 2 weeks after stroke on risk of stroke recurrence at 1-year in a chinese stroke cohort study / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>There were few studies on the relation between changes in libido and incidence of stroke recurrence. The aim of this study was to investigate the relationship between libido decrease at 2 weeks after stroke and recurrent stroke at 1-year.</p><p><b>METHODS</b>It is a multi-centered, prospective cohort study. The 14 th item of the Hamilton Depression Rating Scale-17 was used to evaluate changes of libido in poststroke patients at 2 weeks. Stroke recurrence was defined as an aggravation of former neurological functional deficit, new local or overall symptoms, or stroke diagnosed at re-admission.</p><p><b>RESULTS</b>Among 2341 enrolled patients, 1757 patients had completed follow-up data, 533 (30.34%) patients had decreased libido at 2 weeks, and 166 (9.45%) patients had recurrent stroke at 1-year. Multivariate logistic regression analysis showed that, compared with patients with normal libido, the odds ratio (OR) of recurrent stroke in patients with decreased libido was reduced by 41% (OR = 0.59, 95% confidence interval [CI]: 0.40-0.87). The correlation was more prominent among male patients (OR = 0.52, 95% CI: 0.31-0.85) and patients of ≥60 years of age (OR = 0.57, 95% CI: 0.35-0.93).</p><p><b>CONCLUSIONS</b>One out of three stroke patients in mainland China has decreased libido at 2 weeks after stroke. Decreased libido is a protective factor for stroke recurrence at 1-year, which is more prominent among older male patients.</p>

Carotid artery dissection with Horner syndrome as main manifestation: a case report / 浙江大学学报·医学版

A 44-year old male patient was admitted to the First Affiliated Hospital, Zhejiang University School of Medicine with left ptosis and pain on the left head and neck for 20 days.Brain MRI showed subacute cerebral infarction on left parietal lobe and intramural hematoma on left internal carotid artery. CT angiography showed stenosis line on the C1 segment of left internal carotid artery. Digital subtraction angiography showed dissection on the C1 segment of left internal carotid artery.The condition of patients was improved after anticoagulant therapy.

Anatomical and clinical study of a novel anterial cubital approach for ulnar coronoid fractures / 中华骨科杂志

Objective To investigate the feasibility of a noval anterior cubital approach for the coronoid via flexor?prona?tor teres interval and assess the clinical result. Methods Five formalin?fixed adult cadaver elbows were used. Through a single universal anteromedial longitudinal skin incision, the coronoid tip was exposed via pronator and flexor carpiradialis interval, and coronoid anteromedial facet and base via palm longus and flexor carpi ulnaris interval. The distances from the entry point to the muscles or branching point of the nerves to the line passing through medial and lateral epicondyles, as well as the length were mea?sured with regard to the motor nerve branches arising from median nerve to pronator teres, flexor carpiradialis, palm longus and flexor digiti superficialis, as well as the most proximal two motor branches to flexor carpi ulnaris arising from ulnar nerve. From September 2013 to August 2014, 4 male patients with ulnar coronoid fracture were treated operatively through the above anterior cubital approach in our hospital. They were all left side involved, with an average age of 32 years (range, 16-42 years). According to O’Driscoll classification, there were two cases of type Ib and two cases IIb respectively. They were all treated by open reduction and internal fixation through flexor?pronator teres interval. Results At cubital fossa, there were 2-3 branches to the pronator teres mostly, 1 branch to flexor carpiradialis and palm longus arising from median nerve. The branch to the flexor digiti superficia?lis usually was long and thick, and divided into 2-5 short twigs near muscle. The branch to palm longus had the same trunk with that to flexor digiti superficialis. The branch to flexor digiti superficialis was the most proximal among those passed through the in?terval of pronator teres and flexor carpiradialis, and its entry point to the muscle had an averaged distance of 37.22 mm to the line passing through medial and lateral humeral epicondyles. It was optimal to expose coronoid tip through the interval of pronator teres and flexor carpiradialis. It was safe to expose coronoid proximal to the branch to flexor digiti superficialis. While, it was better to expose the anteromedial facet and base of corocoid through the interval of palm longus and flexor carpiulnaris via median and ul?nar nerve interface. All of the four patients were followed up for an average period of 9 months. They all achieved bone union from 6 weeks to 3months (mean, 9 weeks). All the patients obtained excellent result according to the modified An&Morrey elbow per?formance index with scores from 94 to 100. Conclusion The novel anteromedial cubital approach via flexor?pronator teres is opti?mal for exposure of coronoid.

Intracranial Extramedullary Hematopoiesis in Beta-Thalassemia

Extramedullary hematopoiesis (EMH) represents tumor-like proliferation of hemopoietic tissue which complicates chronic hemoglobinopathy. Intracranial EMH is an extremely rare occurrence. Magnetic resonance imaging (MRI) offers a precise diagnosis. It is essential to distinguish EMH from other extradural central nervous system tumors, because treatment and prognosis are totally different. Herein, we report the imaging findings of beta-thalassemia in a 13-year-old boy complaining of weakness of left side of the body and gait disturbance; CT and MRI revealed an extradural mass in the right temporoparietal region.

The influence on masticatory function after curved osteotomy of prominent mandibular angle / 中华整形外科杂志

<p><b>OBJECTIVE</b>To investigate the influence of mandibular curved osteotomy on masticatory function by analyzing the bite force before and after operation.</p><p><b>METHODS</b>From Jan. to Oct. 2008, 20 female cases underwent mandibular curved osteotomy. The maximal bite force in incisor area, bilateral premolar area and molar area was measured before operation and 6 months after operation.</p><p><b>RESULTS</b>The maximal bite force in incisor area, right premolar area, left premolar area, right molar area, left molar area was (11.7 +/- 3.9) kg, (23.2 +/- 1.6) kg, (30.9 +/- 2.3) kg, (35.6 +/- 4.2) kg, (38.5 +/- 3.1) kg, respectively before operation; and was (11.9 +/- 2.1) kg, (23.0 +/- 4.5) kg (31.0 +/- 1.8) kg,(35.9 +/- 3. 5) kg. (38.5 +/- 2.7) kg, respectively 6 months after operation. The postoperative maximal bite force reached the preoperative level, and even slightly above it. There was no significant difference between the measured data before operation and 6 months afterward (P > 0.05).</p><p><b>CONCLUSIONS</b>The curved osteotomy of prominent mandibular angle has no long-term effect on masticatory function.</p>

Quantitative analysis and significance of CXCL12 and CXCR4 expression with lymphangiogenesis of pancreatic adenocarcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the expression of CXCL12, its receptor CXCR4 and its correlations with clinical pathology and lymphangiogenesis in pancreatic adenocarcinoma (PAC).</p><p><b>METHODS</b>The tissue samples were obtained from 30 patients with PAC by surgery between January 2005 and December 2007, which including PAC, the cancerous peripheral tissues, the normal pancreatic tissues and peripheral lymph nodes. The patients age ranged from 35 to 78 years old (median 57.2 years old). The expressions of CXCL12 and CXCR4 in these tissues were assayed by immunohistochemical staining, RT-PCR and fluorescence quantitative real-time PCR.</p><p><b>RESULTS</b>In the immunohistochemical staining, the CXCL12 protein mainly located in the normal pancreatic cell envelopes and/or cytolymphs. In the immunohistochemical staining, the CXCR4 protein mainly located in the cell envelopes and/or cytolymphs of PAC. The results of RT-PCR and fluorescence quantitative real-time PCR indicated that the expression levels of CXCR4 mRNA in PAC tissues, the cancerous peripheral tissues and peripheral lympho nodes were higher than that in the normal pancreatic tissues (P < 0.01). The MLVD in PAC were detected by morphometric analysis respectively. The level of MLVD in III-IV stages was higher than I-II stages of PAC (P < 0.01), and in these cases which had lymphatic metastasis, the level of MLVD significantly increased (P < 0.01). And there was no correlation between the differentiation and histology types of PAC (P > 0.05). There was 22 samples that the CXCR4 protein was positive, and among these samples the MLVD was higher than that in negative group of CXCR4 protein (P = 0.003).</p><p><b>CONCLUSIONS</b>The expression of CXCR4 was significantly associated with lymphatic metastasis of PAC, and the higher expression of CXCR4 in PAC tissues was significantly associated with lymphangiogenesis of PAC.</p>

Experimental study on gelatin/polycaprolactam composite nanofiber scaffold in wound healing / 中华烧伤杂志

<p><b>OBJECTIVE</b>To explore the effects of the gelatin (Gt)/(polycaprolactam) PCL composite nanofiber scaffold on wound healing of full-thickness defect in rabbits.</p><p><b>METHODS</b>Sixteen rabbits were inflicted with full-thickness skin open wounds on the back, and they were divided into A group (with treatment of Gt/PCL nanofiber membrane, 8 wounds), B group (with treatment of PCL fiber membrane, 8 wounds), and C group (with treatment of vaseline gauze, 8 wounds), the wounds in A , B groups served as self controls. Wound healing time was recorded. Wound healing rates were calculated on 3, 7, 10 post operation day (POD). Wound specimens and their adjacent skin were examined histologically on 3, 7, and 10 POD.</p><p><b>RESULTS</b>There was obvious difference in wound healing time among A, B, C groups, and they were (18.2 +/- 1.3) d, (20.3 +/- 1.1) d, (22.0 +/- 0.6) d, respectively. Wound healing rate in A group was obviously higher than that in B, C group on 3, 7, 10 POD (P < 0.05). Compared with those in other groups, the proliferation of granulation tissue in dermis in A group was less, the epidermal cells were proliferated quickly, and collagen was arranged regularly.</p><p><b>CONCLUSION</b>Gt/PCL composite nanofiber membrane can promote wound healing of full-thickness skin defect in rabbits, which is an ideal material for tissue engineering.</p>

Filter-based template preparation for rapid and sensitive PCR detection of Staphylococcus aureus in meat / 生物工程学报

In the present study, an assay using Polymerase Chain Reaction (PCR) was developed for the detection of Staphylococcus aureus in meat. Based on flotation and solvent extraction technology, FTA filter was used to extract S. aureus DNA from artifically contaminated meat. Primers targeting the thermostable nuclease gene (nuc) were used to amplify a 279 bp DNA fragment which was confirmed by DNA sequencing. The detection limit of PCR was 10 cfu x g(-1) meat of S. aureus. This novel FTA-PCR assay allows for detection of Staphylococcus aureus in meat in <6 h, which is 12-24 h less than that of conventional PCR with enrichment method. Seventy-two samples were analyzed and the detection rate using the standard cultivation method was 70.8%, detection time was 5d. The detection rate of PCR amplification using filters was 73.6%, detection time was 6h. The detection rate of Baird-Parker R.P.F method was 69.4%, detection timewas 18 h. The detection rate of petrfilm RSA method was 61.1%, detection time was 18 h. Thus PCR amplification using filters provides a faster and more sensitive method of S. aureus detection than the standard cultivation method. At the same time,it provides an universal process for preparing DNA template.

ONE NEW SPECIE OF CITEROMYCES FROM CHINA SWEET FLOUR PASTE / 微生物学通报

The investigators isolated one yeast from China Huaimao sweet flour paste. The strain was identified into Citeromyces, and claimed being a new species. The strain was named as Ctieromyces baadingensis zhang sp. Nov., which differed markedly from Citeromyces matritensis in physiological and biochemical characteristics. Citeromyces baod-ingensis didn't ferment sucrose and raffinose, and assimilated galactose and cellobiose, and didn't assimilate galactose and ceflobiose. The G+C mol% was 48.5.