Objeictivie To investigate the diagnostic value of anti-mutated citrullinated vimentin (anti-MCV) antibody, anti-cyclic citrullinated peptide (anti-CCP) antibodies and glucose-6-phospha-teisomerase (GP1) for rheumatoid arthritis (RA). Methods Anti-MCV antibody, GPI and anti-CCP antibody were detected in serum samples of 109 RA patients, 24 non-RA rheumatic diseases patients and 19 healthy blood donors. The sensitivity and specificity of these parameters for the diagnosis of RA were analyzed. Results Both the positive rate and average cut off concentration of anti-MCV and GPI in RA were higher than those of non-RA rheumatic diseases or healthy controls (P<0.05). A significant difference was found between anti-MCV and GPI in RA patients. The most sensitive and specific parameter in RA was anti-MCV (99.1%) and anti-CCP (90.7%) respectively, but, when anti-MCV combined with anti-CCP, or GPI or anti-CCP and GPI, the specificity could be up to 98.1%. Coniclusions Anti-MCV, anti-CCP and GPI alone or in combination may be valuable parameters for the diagnosis of RA.

0.05),there was not statistically different before and after HIFU. However,the normal myometrium after HIFU enhanced obviously just as that before treatment. Conclusion MRI was useful for the evaluation of the intensity, volume changes and the blood supply in leimyomas before and after HIFU.

OBJECTIVE To explore the safe and effective dose of sirolimus (Rapamycin,Sir) and its effect on seizure comorbidities. METHODS Immature C57BL/6 mice at postnatal 10 d of age were administered with kainic acid(KA) 12.0 mg · kg-1 intraperitoneally by a single injection to induce acute seizure. Sir 0.3, 1.0 and 3.0 mg · kg-1 was injected 24 h after seizure every other day until 3 d, 1 week, 3 weeks, 5 weeks and 6 weeks. Western blotting analysis was used to detect the expression and phos?phorylation level of S6 protein and to determine the minimum effective dose of Sir. Effect of the mini?mum effective dose of Sir on cognitive function and body growth was observed by several evaluations. Immunofluorescent intensity of Doublecortin (DCX) immunofluorescent staining was conducted to evaluate the development of neurons in the hippocampus. Morris water maze was used to assess the cognitive function. Tail suspension test, O maze and new object recognition test were used to study the anxiety-like behaviors of mice. RESULTS The result of Western blotting showed that Sir 0.3 mg · kg-1 had no significant effect on the phosphorylation of S6 protein in normal mice or KA mice, whereas 1.0 and 3.0 mg · kg- 1 could significantly inhibit the phosphorylation of S6 protein in KA mice (P<0.05). Sir 1.0 mg·kg-1 had no obvious effect on DCX-positive cells or body wass. Morris water maze showed that KA-induced seizure resulted in prolonged escape latency and swimming length (P<0.05), and a decreased crossing number of target quadrant (P<0.05). Sir 1.0 mg·kg-1 significantly reversed the deficit of cognitive function of KA-induced seizure mice (P<0.05), whereas no significant difference was found between Sir group and normal control group. Compared with normal control group, model group showed increased freezing time in tail suspension test (P<0.05), decreased migration length and reten?tion time in open arms in O maze (P<0.05), decreased retention time and touch frequency with new objects, migration length and average speed in new object recognition test (P<0.05). Sir 1.0 mg · kg-1 significantly reversed the above anxiety and depression status, whereas no significant difference was found between sirolimus group and normal control group. CONCLUSION Sir 1.0 mg · kg-1 inhibits the abnormal activation of mTOR pathway and the formation of epilepsy comorbidity in immature mice. Along with its mild side effect in development, Sir 1.0 mg · kg-1 will be an ideal dose to be used in the treatment of seizure in immature mice.

Objective:To investigate the expression of Slug,EMMPRIN and E-cadherin in salivary adenoid cystic carcinoma (SACC)and its correlation with clinicopathological characteristics,and the correlation among themselves.Methods:Slug,EMMPRIN and E-cadherin expression in 1 1 5 SACC cases of SACC was examined by immunohistochemical staining.The results and clinicopatho-logical data were statistically analyzed.Results:High positive expression frequencies of Slug(76.5%)and EMMPRIN(69.6%)and low positive expression frequency of E-cadherin(51 .3%)were found in 1 1 5 SACC cases.The expression of Slug and EMMPRIN was positively associated with the histopathological types,clinical stages,perineural invasion,recurrence and distance metastasis(P <0.05).The expression of E-cadherin was negatively associated with the histopathological types,clinical stages,perineural invasion and distance metastasis(P <0.05).There was a significant correlation between Slug and EMMPRIN expression(P <0.05),negative correlation between EMMPRIN and E-cadherin expression(P <0.05)and between Slug and E-cadherin expression(P <0.05).Con-clusion:The expression of Slug,EMMPRIN and E-cadherin is closely correlated to the clinicopathological characteristics of SACC.

BACKGROUND:Distraction osteogenesis is one of the most important tissue engineering technologies. However, the exact signaling pathway controling mesenchymal stem cel-osteoblast lineage (MSC-OB) migration during distraction osteogenesis has not yet been elucidated. More efforts should be paid to make a ful understanding of the mechanism on MSC-OB lineage migration, which can improve the clinical efficacy of distraction osteogenesis.
OBJECTIVE:To evaluate the effects of mechanical stretch on the ability of MSC-OB mobility and expression of mammalian target of rapamycin (mTOR) signaling pathway as wel as matrix metaloproteinases (MMPs) in MSC-OB, and to make clear the mechanism by which controls MSC-OB migration during distraction osteogenesis.
METHODS:Twelve Sprague-Dawley rats were randomized into two groups: experimental group (n=6), anin vivo rat mandibular distraction osteogenesis model was established on the right side of rats; non-stretch group (n=6), only the mandibular resection was done but with no distraction osteogenesis. Immunohistochemical staining was used to detect phosphorylated mTOR expression in new osteotylus at 15 days after operation. In addition, an in vitro cel stretch model was made in the mandibular mesenchymal stem cels from healthy Sprague-Dawley rats under resting tension force (6%, 4 hours); no distraction was done in control group. The ability of MSC-OB mobility, the expression of mTOR, Raptor, p70S6K and MMPs were evaluated using experiment methods including immunohistochemistry staining, real-time PCR and scratch assay.
RESULTS AND CONCLUSION: The expression of phosphorylated mTOR in MSC-OB was upregulated in the mandibular bone calus of the stretch group than the non-stretch group (P < 0.05). In thein vitro experiments, MSC-OB applied with mechanical stretch (6%, 4 hours) showed elevated gene expression levels of mTOR, Raptor, p70S6K, MMP-2, MMP-9 and MMP-13 compared with the control group (0%, 4 hours). Meanwhile, MSC-OB in the experiment group (6%, 4 hours) showed a greater ability of mobility, as demonstrated by a farther distance after 48 hours of observation (P < 0.05). The present study suggests that the enhancement of MSC-OB mobility correlates with increase of the gene expression of MMPs and mTOR signaling pathway. Mechanical stretch may promote MSC-OB migration through activation of mTOR/MMPs signaling pathway.

OBJECTIVETo study functions of Jingu Tongxiao granule (JGTXG, treatmenting ache of bones and muscles) in antiphlogistic and antalgic aspect, invigorating the circulation of blood and absorbing clots and antitraumatic soft tissue.

METHODAnimal models of inflammation, ache, gore and traumatic soft tissue were adopted, and pharmacodynamic actions of Jingu Tongxiao granule were observed.

RESULTJGTXG could conspicuously restrain inflammatory reactions of mouse ear tumid model treated by croton oil tumid and rat foot metatarsus tumid model treated by carrageenan, and restrain pain responses of mouse caused with whipping back end method by heat stimulating and of mouse caused with wriggling body method by acetic acid being injected in its abdominal cavity. It could significantly improve petechia degree in traumatic rat blood stasis model, and prominently improve raumatized limb's tumefaction degree and alleviate blood stasis, swelling and phlogistic cell soakage in traumatic rat soft tissue model. At the same time, it could prominently restrain platelet aggregation and improve whole blood viscosity.

CONCLUSIONJingu Tongxiao granule has antiphlogistic and antalgic functions, invigorating the circulation of blood and absorbing clots and antitraumatic soft tissue, and it could keep curative effect of original dosage form.

Analgesics, Non-Narcotic ; pharmacology ; Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; Blood Viscosity ; drug effects ; Cinnamomum ; chemistry ; Cyperus ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Ear Diseases ; pathology ; Edema ; pathology ; Hemorheology ; drug effects ; Male ; Mice ; Pain Threshold ; drug effects ; Plants, Medicinal ; chemistry ; Platelet Aggregation ; drug effects ; Rats ; Rats, Sprague-Dawley ; Salvia miltiorrhiza ; chemistry

Seventy patients with advanced non-small-cell lung cancer (NSCLC) aged 65 or above were treated with epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI) erlotinib or gefitinib from February 2006 to September 2010. The efficacy and toxicities of treatment were retrospectively analyzed.The overall response rate and disease control rate were 31.4％ and 84.3％,respectively. Themedian progression-free survival time and median survival time were 8.0 months and 13.5 months,respectively(P ＜ 0.05 ). One-year survival rate was 54.3％. Response rate ( CR + PR) ( 42.9％ ) anddisease control rate (94.3％ )in female patients were superior to males (20.0％ and 74.3％ ) (P ＜ 0.05 ).Non-smoking and PS score ＜ 2 were good predictors for survival.The side effects were generally mild and mainly were skin rash and diarrhea.

Objective:To evaluate the effect of tubastatin A (TubA) on pyroptosis during brain injury after cardiac arrest and resuscitation in swine.Methods:Twenty-two conventional male white swine, weighing 34-39 kg, aged 4-6 months, were divided into 3 groups using a random number table: sham operation group (group S, n=6), cardiac arrest-cardiopulmonary resuscitation (CA-CPR) group ( n=8) and CA-CPR+ TubA group ( n=8). The swine model of CA-CPR was established by 9 min of cardiac arrest and 6 min of cardiopulmonary resuscitation in CA-CPR group and CA-CPR+ TubA group. TubA 4.5 mg/kg (in 50 ml of normal saline) was infused over 1 h via the femoral vein starting from 5 min after resuscitation in CA-CPR+ TubA group. Before developing the model and at 1, 2, 4 and 24 h after resuscitation (T 0-4), blood samples were collected from the femoral vein for determination of the concentrations of neuron specific enolase (NSE) and S100β protein in serum (by enzyme-linked immunosorbent assay). Neurological deficit score (NDS) was evaluated at T 4. The animals were then sacrificed, and their brain cortex tissues were harvested to measure the expression of histone deacetylase 6 (HDAC6), caspase-3, cleaved caspase-3, gasdermin E (GSDME) and GSDME N-terminal (N-GSDME) (by Western blot) and contents of high mobility group box 1 (HMGB1), interleukin-1β (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay). Results:Compared with group S, the serum concentrations of NSE and S100β were significantly increased at T 1-4, NDS was increased at T 4, the expression of HDAC6, caspase-3, cleaved caspase-3, GSDME and N-GSDME in brain cortex was up-regulated, and the contents of HMGB1, IL-1β and IL-18 were increased in CA-CPR and CA-CPR+ TubA groups ( P<0.05). Compared with group CA-CPR, the serum concentrations of NSE and S100β were significantly decreased at T 3, 4, NDS was decreased at T 4, the expression of HDAC6, caspase-3, cleaved caspase-3, GSDME and N-GSDME in brain cortex was down-regulated, and the contents of HMGB1, IL-1β and IL-18 were decreased in group CA-CPR+ TubA ( P<0.05). Conclusions:The mechanism by which TubA alleviates brain injury after cardiac arrest and resuscitation may be related to inhibition of pyroptosis in swine.

Objective:To investigate the effect of LRRK2G2019S mutation on activation of microglia after iron deprivation and its mechanism.Methods:(1) Microglia were differentiated from human induced pluripotent stem cells (IPSC) with the help of hematopoietic progenitor cells (HPC) and identified by immunofluorescent staining, and α-synuclein (α-syn) A53T mutant protein was obtained by protein purification technology. (2) Microglia were divided into control group, α-syn group, α-syn+ deferoxamine (DFO) group; phosphate buffer solution (PBS), 1 μmol/L purified α-syn A53T mutant protein, 1 μmol/L purified α-syn A53T mutant protein+30 mmol/L DFO were given respectively for 24 h. Fe 2+ concentration was detected by colorimetry, Rab35 protein expression was detected by Western blotting, intracellular reactive oxygen species (ROS) level was detected by flow cytometry, and interleukin-6 ( IL-6), tumor necrosis factor-α ( TNF-α) and transforming growth factor-β ( TGF-β) mRNA expressions were detected by real time-PCR (RT-PCR); microglia culture supernatant (MCS) in the 3 groups were transfered to SH-SY5Y cells, and SH-SY5Y cell apoptosis was detected by flow cytometry. (3) Bidirectional DNA sequencing was used to detect leucine rich repeat kinase 2 ( LRRK2) gene mutations in microglia treated with 1 μmol/L purified α-syn A53T mutant protein. Microglia were divided into control group, α-syn group and α-syn+GSK3357679A group, and treated with corresponding drugs for 24 h, respectively (LRRK2 inhibitor GSK3357679A concentration: 10 nmol/L), and LRRK2 protein expression was detected by Western blotting; microglia were divided into control group, α-syn group, α-syn+GSK3357679A, and α-syn+GSK3357679A+DFO group, and treated with corresponding drugs for 24 h, Rab35 protein expression was detected by Western blotting, intracellular ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. (4) Microglia were divided into control group, α-syn group, α-syn+rapamycin (RAPA) group, and treated with corresponding drugs for 24 h (concentration of autophagy inducer RAPA: 50 nmol/L); protein expressions of Rab35, P62 and microtubule-associated protein light chain 3 II (LC3II) were detected by Western blotting; intracellular ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. (5) Microglia were divided into control group, α-syn group, and α-syn+Rab35 group, and treated with corresponding drugs for 24 h (concentration of Rab35 overexpressed plasmids: 1 μg/mL); Rab35, P62, and LC3II protein expressions were detected by Western blotting; ROS level was detected by flow cytometry, and IL-6, TNF-α and TGF-β mRNA expressions were detected by RT-PCR. Results:(1) Immunofluorescent staining showed negative neuronal nuclei (NeuN) expression and positive ionized calcium-binding adapter molecule 1 (Iba1) expression in microglia, and high LRRK2 expression; PcDNA3.1-SNCA-A53T expression plasmid was constructed and α-syn A53T mutant protein was purified. (2) The Fe 2+ concentration in α-syn group was significantly higher than that in control group, and the Fe 2+ concentration in α-syn+DFO group was significantly lower than that in α-syn group ( P<0.05); the Rab35 protein and TGF-β mRNA expressions in control group, α-syn group and α-syn+DFO group were decreased successively, while the IL-6 and TNF-α mRNA expressions were increased successively, with significant differences ( P<0.05); ROS level and SH-SY5Y cell apoptosis rate in control group, α-syn group, α-syn+DFO group were increased successively. (3) Bidirectional DNA sequencing showed that the LRRK2G2019S mutation in microglia was the most obvious after α-syn A53T mutant protein stimulation; compared with the control group, the α-syn group had significantly increased LRRK2 protein expression, while the α-syn+GSK3357679A group had significantly decreased LRRK2 protein expression compared with α-syn group ( P<0.05); compared with the control group, the α-syn group had significantly decreased Rab35 protein and TGF-β mRNA expressions, and statistically increased IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn group, the α-syn+GSK3357679A group had significantly increased Rab35 protein and TGF-β mRNA expressions, and statistically decreased IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn+GSK3357679A group, α-syn+GSK3357679A+DFO group had significantly increased IL-6 and TNF-α mRNA expressions, and significantly decreased Rab35 protein and TGF-β mRNA expressions ( P<0.05). The α-syn group had higher ROS level than the control group, the α-syn+GSK3357679A group had lower ROS level than the α-syn group, and the α-syn+GSK3357679A+DFO group had higher ROS level than the α-syn+GSK3357679A group. (4) Compared with the control group, the α-syn group had significantly decreased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly increased P62 protein, IL-6 and TNF-α mRNA expressions ( P<0.05); compared with α-syn group, the α-syn+RAPA group had significantly increased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly decreased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05); the α-syn group had higher ROS level than the control group and α-syn+RAPA group. (5) Compared with the control group, the α-syn group had significantly decreased Rab35 and LC3II protein, and TGF-β mRNA expressions, and statistically increased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05); compared with the α-syn group, the α-syn+Rab35 group had significantly increased Rab35 and LC3II protein, and TGF-β mRNA expressions, and significantly decreased P62 protein, and IL-6 and TNF-α mRNA expressions ( P<0.05). The α-syn group had higher ROS level than the control group and α-syn+Rab35 group. Conclusion:LRRK2G2019S can induce neuroinflammation by inhibiting Rab35-related autophagy under iron deprivation, and Rab35 is expected to be a key factor in intervening neuroinflammation.

Objective:To evaluate clinical characteristics and treatment of postoperative anastomotic stricture in pediatric congenital biliary dilatation patients.Methods:The clinical data of 24 children with postoperative anastomotic stricture from Apr 2012 to Oct 2019 in Beijing Children's Hospital was retrospectively analyzed.Results:There were 6 males and 18 females. Patients were divided into bile- leak group (BL, n=6) and non bile-leak group (NBL, n=18) based on whether there was anastomotic leakage after primary surgery. The main symptoms in BL group was persistent obstructive jaundice, and recurrent cholangitis in NBL group. Postoperative symptoms were first shown in an average of 7.0 months in BL group, compared to 59.0 months in NBL group, P<0.05. In BL group, 4 underwent redoing hepaticojejunostomy, 2 underwent anastomosis plasty. In NBL group, 3 underwent redoing hepaticojejunostomy, 15 did anastomosis plasty with multiple biliary stones found necessitating extraction. After reoperation, one patient had bile leakage, 2 patients had recurrent cholangitis within one-month, 21 patients had uneventful recovery. Five were found to have biliary stones in long-term follow-up. Conclusions:Biliary-enteric anastomotic leakage can cause stricture in postoperative patients of congenital biliary dilatation ,reoperation is necessary in symptomatic patients.