Objective To investigate the effect of Shufeng Xuanfei and Jiebiao Qingli concoctions on Toll-like receptor (TLR) signal pathway of pneumonia infected with influenza virus in mice.Methods The pneumonia model was reproduced by nasal dropping of influenza virus A in mice.The mice were randomly divided into nine groups:normal group (C),model group (M),tamiflu group (D),Shufeng Xuanfei low-dose (SL),medium-dose (SM) and high-dose (SH) groups,Jiebiao Qingli low-dose (JL),medium-dose (JM) and high-dose (JH) groups,each n =12.Two hours after model-reproduction,the mice in C group and M group received distilled water by gavage.The mice in D group received 2.5 g· mL-1· d-1 oseltamivirphosphate.Shufeng Xuanfei formula in doses of 3.76,1.88,0.94 g· kg1 · d-1 were respectively administered to SH,SM and SL groups by gavage,Jiebiao Qingli formula in doses of 4.37,2.18,1.09 g ·kg-1 ·d-1 was given to JH,JM and JL groups by gavage,respectively.Each group was in equal dose of 0.2 mL daily over a 4-day period.Total RNA was extracted in each group.Then gene chips were used to screen these RNA samples.Some genes that were involved in TLR signal pathways were selected.These candidate genes were verified by real-time reverse transcription-polymerase chain reaction (RT-PCR).Results TLR7,MYD88,CCLS,IFNB1,IL6,IL12a,NFKBIA and IKBKB were up-regulated in model group compared with control group.Compared with model group,down-regulated genes in medium-dose,low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula included TLR3,TLR7,MYD88,CCL5,IFNB1,IL6,IL12a,NFKBIA and IKBKB (log2 signal intensity of SM,/M in medium-dose Shufeng Xuanfei formula group were-1.24,-2.02,-1.36,-1.95,-0.63,-1.33,-3.50,-1.33,-1.33,log2 signal intensity of SI/M in low-dose Shufeng Xuanfei group were-1.07,-2.43,-2.63,-2.30,-5.09,-3.19,-3.53,-1.95,-1.95,log2 signal intensity of JM/M in medium-dose Jiebiao Qingli formula group were -1.78,-0.55,-1.35,-1.47,-1.65,-2.03,-3.02,-1.57,-1.57,respectively).The results suggested that the effect of Shufeng Xuanfei formula was better than that of Jiebiao Qingli formula.By RT-PCR,compared with model group,low-dose,medium-dose and high-dose groups of Shufeng Xuanfei formula,medium-dose and high-dose groups of Jiebiao Qingli formula,and tamiflu group,significant decrease in TLR7,nuclear factor-κB (NF-κB),myeloid differential protein-88 (MyD88) mRNA expression were found.Medium-dose and low-dose Shufeng Xuanfei formula group (TLR7 mRNA:3.6 ±0.3,3.5 ± 1.2 vs.7.4 ± 1.6,NF-κB mRNA:1.1 ±0.2,1.0 ±0.2 vs.2.2 ±0.4; MyD88mRNA:1.4 ± 0.4,1.0 ± 0.3 vs.3.4 ± 0.9,all P＜0.01) and medium-dose Jiebiao Qingli formula group (TLR7 mRNA:4.9 ± 0.3 vs.7.4 ± 1.6,NF-κB aRNA:1.3 ± 0.7 vs.2.2 ± 0.4,MyD88 mRNA:1.6 ± 0.8 vs.3.4 ± 0.9,P＜0.05 or P＜ 0.01) were shown statistically significant decreases compared with the model group.Conclusions Medium-dose and low-dose Shufeng Xuanfei formula and medium-dose Jiebiao Qingli formula can inhibit the inflammatory reaction induced by influenza virus by down-regulating the NF-κB through TLR signal pathways dependent on MyD88.The regulation of Shufeng Xuanfci formula in TLR signal pathways was superior to that of Jiebiao Qingh formula.

This study was aimed to clone the GGPS (geranylgeranyl pyrophosphate synthase) gene from Lepidium apetalum, to analyze its sequence, and to express the protein in E.coli expression system. Specific PCR cloning primers were designed for GGPS gene from Lepidium apetalum according to the full-length sequence from a previous transcriptome sequencing project. PCR amplification was performed with this primer pair on a leaf cDNA template. TA cloning, sequencing and sequence analysis were performed.GGPS gene from Lepidium apetalum was expressed in the E.coli expression system. The results showed that the full-lengthGGPS cDNA from Lepidium apetalum was 1 146 bp coding a protein of 381 amino acids. The LaGGPS protein had an isoprenoid synthase domain. According to a phylogenetic tree constructed with multiple alignment of GGPS protein sequences from various plant species, GGPS protein from Lepidium apetalum was the closest to Arabidopsis thaliana and Sinapis alba. The prokaryotic expression vectorpET-32a-LaGGPS was also constructed successfully. The protein was expressed in E.coli BL21 strain. It was concluded that the cloning and prokaryotic expression of LaGGPS gene provided a foundation for a follow-up research of its function with protein purification and activity analysis.

Objective To prospectively investigate the characteristics and correlative influential factors of pulmonary hypertension (PHT) in patients on long-term maintenance hemodialysis (MHD). Methods Pulmonary artery systolic pressure (PASP) was assessed by echocardiography according to the guideline from the American Society of Echocardiography in 2010 and PASP more than 35 mm Hg was diagnosed as PHT.Echocardiography and pulse wall velocity (baPWV) was performed in the next day after hemodialysis.Arteriovenous fistula (AVF) flow was evaluated by the ultrasound dilution method.Hemodialysis-related informations and laboratorial parameters were detected in the same period. Results One hundred and eleven MHD patients [male 45,female 66,mean age (57.32±12.49) years old] in our hemodialysis center were included in the study.All of the patients received MHD treatment for more than 6 months with AVF as the vascular access.The patients with any possible diseases causing PHT were excluded.The mean MHD period was (70.51±44.98) months.Twenty-eight patients (25.32％) were diagnosed as PHT with mean PASP (45.68±10.83) mm Hg.Left ventricular diastolic dysfunction was severer in patients with PHT than that in patients without PHT.The prevalence of moderate to severe diastolic dysfunction was statistically higher in PHT group compared to non-PHT group (53.60％ vs 6.02％,P＜0.01).Ejection fraction (EF),fractional shortening of left ventricular diameter in PHT group were also significantly lower than those in non-PHT patients (62.06％±14.90％ vs 69.72％±8.60％,36.46％±10.04％ vs 40.20％±7.86％,P＜0.01).The patients with EF less than 50％ were 21.43％and 3.61％ in PHT and non-PHT group respectively.However,there were no significant differences in age,sex,MHD periods,body mass index (BMI),interval dialysis weight growth,blood pressure before dialysis,hemoglobin,albumin,pre-albumin,serum calcium and phosphorus,iPTH,nPCR,Kt/V,baPWV and AVF flow between the two groups. Conclusions PHT is a common complication of patients on long-term MHD.There is close relationship between PHT and left ventricular insufficiency.PHT is not significantly relevant to mineral metabolic disturbance,AVF flow,hemoglobin,dialysis adequacy and nutrition status.

BACKGROUND:Conflicting data have been reported regarding the expression of retinol-binding protein 4 in non-alcoholic fatty liver disease. OBJECTIVE:To evaluate the impact of dietary interventionversus metformin treatment on expression of retinol-binding protein 4 in rats with non-alcoholic fatty liver disease. METHODS: Fifty male Sprague-Dawley rats were randomized to six groups, including two normal control groups (rats were kiled after 8 and 16 weeks of normal diet), two HFD groups (rats were kiled after 8 and 16 weeks of high-fat diet), one dietary intervention group (rats were kiled after 8 weeks of high-fat diet and 8 weeks of normal diet) and one metformin treatment group (rats were kiled after 8 weeks of high-fat diet and 8 weeks of high-fat diet and metformin treatment). The levels of retinol-binding protein 4 in serum and biochemical indexes were detected through enzyme-linked immunosorbent assay. The expression of retinol-binding protein 4 mRNA in liver tissues was measuredvia western blot analysis, immunohistochemistry and RT-PCR. RESULTS AND CONCLUSION:Non-alcoholic fatty liver disease models were successfuly established by high-fat diet. Liver tissues of high-fat diet fed rats showed progressing non-alcoholic fatty liver disease histology, from non-alcoholic fatty liver to non-alcoholic steatohepatitis. Dietary intervention increased retinol-binding protein 4 expression in liver tissue as wel as improving liver enzyme, dyslipidemia and insulin resistance, and aleviated impaired liver histology. Metformin treatment only aleviated hepatic steatosis caused by high-fat diet. The results indicated that retinol-binding protein 4 expression might play a role in the pathogenesis of non-alcoholic fatty liver disease. Metformin treatment can aleviate non-alcoholic fatty liver disease histology,dietary intervention should be the fundamental treatment.

Objective To evaluate the safety,effectiveness,and middle or long-term efficacy of endovascular stenting of internal carotid artery stenosis at the cavernous segment. Methods Thirty-two patients underwent endovascular stenting at the cavernous segment of internal carotid artery from January 2012 to February 2015 were enrolled retrospectively. Angioplasty and stenting were conducted using Apollo or Winspan stent system. The improvement of internal carotid artery cavernous segment stenosis and perioperative safety and the results of the medium and long-term follow-up of the 2 kinds of stents were observed. Results All the 32 patients achieved technical success. The symptoms of cerebral ischemia of the patients were relieved significantly. The length of the stenosis at cavernous segment of the internal carotid artery was 4 to 13 mm (mean,7. 2 ±2. 9 mm). The stenosis rate from 82 ± 7% before treatment decreased to the 24 ± 7% . One patient had perioperative complication (4. 7%),26 of them were followed up with DSA,and 6 were lost to follow-up. The follow-up period ranged from 7 to 29 months (mean,16 ± 7 months). During the follow-up period,1 patient had intracerebral hemorrhage,1 had cerebral infarction,and none of them died. Four patients had in-stent restenosis,three of them used Winspan stents, and 1 used Apollo stents. Conclusion The patients should be screened strictly,particularly paying attention to the length of lesions. Endovascular stent angioplasty for the treatment of internal carotid artery cavernous segment stenosis is a safe and effective method.

Objective To observe the therapeutic effect on the treatment of avascular necrosis of femoral head by core decompression with autograft of mesenchymal stem cells(MSCs).Methods Nineteen patients with avascular necrosis of femoral head,including 8 cases of phase I,6 cases of phase II and 5 cases of phase III based on ARCO classification,were treated with core decompression with autograft of MSCs.Results The patients were followed up for one year.The average ratio of necrosis zone diminished from 31.88% to 13.18%.The Harris hip score was increased significantly.Conclusion The treatment of core decompression with autograft of MSCs is safe and effective,and it deserves to application in early necrosis of femoral head.

ObjectiveTo investigate the regulatory effects of traditional Chinese medicine (TCM) Shufengxuanfeijiedu formula on Janus kinase signal transducer and activators of transcription (JAK-STAT) of lung tissues in mice with influenza viral pneumonia.Methods According to random number table, 60 mice were randomly divided into six groups with 10 mice in each group: normal group (N), model group (M), Tamiflu control group (C) and low (SL), medium (SM), high dose (SH) Shufengxuanfeijiedu formula groups. The mouse model of influenza virus pneumonia was reproduced by dropping of 0.05 mL 4LD50 inflluenza virus FM1 strain which can be adapted to lung tissue into the nose; while the N received nose instillation of 0.05 mL normal saline. After successful modeling for 2 hours, distilled water was given orally (by lavage) to N and M; Duffy (oseltamivir) 2.5 g·mL-1·d-1 was administrated to C; the TCM SL, SM, SH were intragastrically administered with different doses of shufengxuanfeijiedu decoction into the corresponding groups respectively (the ingredients of prescription: chrysanthemum, mulberry leaf, almond, platycodon root, forsythia, bupleurum etc. forming granules), according to the suitable dose of granules used for human body surface, the dose used for mouse surface area was calculated, the high dose means the dose used in the medium dose group doubled, the low dose means 1/2 dose used in medium group, once a day, once 0.2 mL for consecutive 4 days. Afterwards, the lung tissues were collected, the mouse differential gene expressions related to JAK-STAT pathway were detected by gene chip technology, the standards for screening of differential gene expression were as follows: up-regulated gene was P < 0.05, and the log2ratio > 1; down-regulation gene wasP < 0.05, and log2ratio < -1. The levels in lung tissue kinase (JAK) andγinterferon (IFN-γ) mRNA expressions were determined by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-qPCR).Results Compared with those in N, the differential expression gene transcription activator, STAT5 [log2 (N/M) = 2.32], interleukin 4 receptor alpha subunit [IL4RA, log2 (N/M) = 4.77], interleukin 12 receptor [IL12R, log2 (N/M) = 1.58], JAK [log2 (N/M) = 2.41] were all obviously up-regulated, and IFN was significantly down-regulated [log2 (N/M) = -1.45] in M. Compared with those in M, C group IFN [log2 (C/M) = 1.51], various TCM dose groups [log2 (SL/M) = 1.46, log2 (SM/M) = 1.72, log2 (SH/M) = 1.40] differential expression gene IFN was significantly up-regulated, STAT5 [log2 (C/M) = -2.06, log2 (SL/M) = -1.41, log2 (SM/M) = -2.10, log2 (SH/M) = -1.89], IL4RA [log2 (C/M) = -2.52, log2 (SL/M) = -1.85, log2 (SM/M) = -2.74, log2 (SH/M) = -1.39), IL12R [log2 (C/M) = -1.48, log2 (SL/M) = -0.10, log2 (SM/M) = -1.58, log2 (SH/M) = -0.53], JAK [log2 (C/M) = -1.44, log2 (SL/M) = -0.88, log2 (SM/M) = -1.74, log2 (SH/M) = -0.53] were significantly down-regulated. In M, the JAK mRNA expression was obviously elevated (2-ΔΔCt: 3.17±0.94 vs. 1.01±0.13,P < 0.05), while the IFN-γ mRNA expression was decreased (2-ΔΔCt: 0.15±0.48 vs. 1.01±0.12,P < 0.05); compared with M, the JAK mRNA expressions in C, SM and SH groups were all obviously decreased (2-ΔΔCt: 2.02±0.63, 1.19±0.30, 1.59±0.67 vs. 3.17±0.94, allP < 0.05); while the IFN-γmRNA expressions in C, SL, SM and SH groups were elevated (2-ΔΔCt: 0.61±0.12, 0.41±0.13, 0.85±0.14, 0.78±0.20 vs. 0.15±0.48, allP < 0.05).Conclusions Shufengxuanfeijiedu formula can ameliorate the mice immune pathological injury of lung tissues induced by influenza virus by regulating JAK-STAT signal pathway and balancing Th1/2 via up-regulating the expression of IFN-γ.

Objective:To evaluate the safety and feasibility of endovascular recanalization for non-acute internal carotid artery occlusion (NA-ICAO), and to propose a new angiographic classification.Methods:From April 2015 to October 2019, 95 consecutive patients with symptomatic NA-ICAO who received endovascular recanalization were retrospectively analyzed in Beijing Tiantan Hospital, Capital Medical University. All the patients were divided into four groups according to DSA: type Ⅰ, petrous segments were distally reconstituted by collateral vessels; type Ⅱ, cavernous segments were distally reconstituted by collateral vessels; type Ⅲ, ophthalmic segments were distally reconstituted by collateral vessels; type Ⅳ, communicating segments were distally reconstituted by collateral vessels. Study data including clinical characteristics, surgical details, lesion classification, recanalization rate and perioperative complications. For the counting data, the χ 2 test was used to compare between groups. For the quantitative data, the ANOVA was used for the normal distribution data, otherwise the Kruskal-Wallis H test was used. The primary safety outcome was any stroke or death within 30 days. Results:Among the 95 patients, 67 (70.53%) had successful recanalization. The recanalization rates of type Ⅰ－Ⅳ were 92.31% (36/39), 81.82% (18/22), 47.83% (11/23) and 18.18% (2/11) respectively (χ2=29.557, P<0.001). And the complication rates of the four types were 5.13% (2/39), 13.64% (3/22), 21.74% (5/23) and 9.10% (1/11) respectively. The incidence of perioperative ischemic stroke was 2.11% (2/95). No other serious stroke and death occurred. Conclusions:Endovascular recanalization may be feasible and safe for carefully selected patients with NA-ICAO and therefore represents an alternative treatment. The patients with type Ⅰ and Ⅱ lesions had higher recanalization rates, while the patients with type Ⅳ lesions had significantly lower recalculation rate. The new angiographic classification is conducive to the selection of suitable patients and difficulty in grading.

Biliary atresia is a severe chronic liver disease in infancy that manifests as destructive inflammatory obstructive cholangiopathy.Recent evidence supports that early Kasai procedure by 30 days of life achieves the greatest likelihood of slowing or preventing the need for liver transplant in biliary atresia patients.Unfortunately,the average age at the time of Kasai procedure is around 60 days of life both in China and abroad.The primary reason for this is that there are many difficulties and challenges in early diagnosis.For example,age stratification should be considered when using GGT levels as a screening tool for biliary atresia in infant less than 30 days.What's more,imaging modality such as US shows lower accuracy in very early stage due to milder pathological change in biliary tract,which post challenges on precision medicine and improving prognosis.This article reviewed different early diagnostic methods of screening for biliary atresia,so as to provide a reference for the early diagnosis of biliary atresia,and help discuss the potential development prospect and research direction in the future.

Objectives To investigate the image quality,influencing factors and radiation doses of prospectively ECG-triggered spiral acquisition mode (Flash spiral mode)coronary computed tomography angiography (CCTA) using high-pitch dual-source CT in patients with different heart rates.Methods One hundred and thirty-four consecutive patients with mean heart rate (HR) ＞ 65 beats per minute (bpm) and ≤ 100 bpm were included in this study as group A using Flash spiral mode setting at 20％ -30％ of the R-R interval and 134 consecutive patients with mean HR ≤65 bpm were enrolled as group B using Flash spiral scan mode at 55％ of the R-R interval; 134 consecutive patients with mean HR ＞ 65 and≤100 bpm using spiral scan mode were included as group C. The image quality scores, effective radiation dose and influencing factors of image quality in three groups were assessed. All statistical analyses were performed using SPSS.Results (1)The non-diagnostic coronary artery segments in group A (28/1842,1.52％) were more than in group B (8/1819,0.44％ ) ( x2 =10.97,P =0.001 ) and there was no significant difference between group A and group C (32/1838,1.74％) ( x2 =0.280,P =0.345).The number of patients with non-diagnostic coronary segments in group A( 10/134,7.5 ％ )was more than in group B (2/134,1.5 ％,x2 =5.52,P =0.018 ),while there was no significant difference between group A and group C (9/134,6.7％,x2 =0.057,P =0.812).(2)The average heart rate variability (HRV) of patients with different image scores in the three groups wcrc significantly different. In group A,the HRV of score 1,2 and 3 were (2.29 ± 1.06),(5.17 ± 1.37),(8.88 ± 1.53) bpm,respectively (F =170.402,P =0.001 ).In group B were (2.26±1.01),(5.97 ±1.82),(12.00 ±9.64) bpm,respectively (F=95.843,P=0.001).In group C were (2.61 ±1.85),(7.90 ±3.97),(11.22 ±5.62) bpm,respectively (F=68.629,P=0.001 ). (3) The average effective radiation doses in groups A and B were significant lower than in group C [ A group was 1.04 ± 0.16,B group was 1.03 ± 0.16 and C group was 7.05 ± 1.05,t =65.5 ( A vs C),P ＜0.01 ( A vs C) ].Conclusions Flash spiral mode of high-pitch dual-source provides high image quality with significant reduction of radiation exposure in patients with HR ≤ 65 bpm. Patients with heart rates ＞65 bpm and ≤100 bpm without cardiac arrhythmia can perform CCTA using Flash spiral mode with image acquisition time setting at 20％-30％ of the R-R interval.