Objective To investigate the effect ofbiglycan (BGN) on neural apoptosis in mice with early brain injury (EBI) after subarachnoid hemorrhage (SAH).Methods SAH models were induced by endovascular perforation in young male C57BL/6J mice.(1) Totally,48 mice were randomly divided into sham-operated group,SAH 6 h group,SAH 12 h group,SAH 24 h group,SAH 48 h group,and SAH 72 h group (n=8);the BGN protein and mRNA expressions were detected by Western blotting and real-time quantitative PCR (qRT-PCR).(2) Totally,16 mice were randomly divided into sham-operated group and SAH 48 h group (n=8);immunofluorescent double staining was conducted to explore the BGN expression in the neurons of brain tissues.(3) Totally,24 mice were randomly divided into sham-operated group,sham+control lentivirus group,and sham+BGN lentivirus group (n=8);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before sham-operation;qRT-PCR was performed to explore the BGN mRNA expression.(4) Totally,48 mice were randomly divided into sham-operated group,SAH+control lentivirus group,and SAH+BGN lentivirus group (n=16);BGN lentiviral vector and control lentivirus were administered intracerebroventricularly 7 d before SAH;neurological scores were detected by modified Garcia scale and beam balance tests;TUNEL was used to detect the neuronal apoptosis,and Western blotting was performed to explore the expressions of nuclear transcription factor kappa B (NF-κB) and phosphorylated-(p-) NF-κB.Results (1) Mice in the SAH 48 h group had the highest BGN protein and mRNA expressions,which showed statistical differences as compared with the sham-operated group (P＜0.05).(2) A majority of BGN expressions were detected in the neurons 48 h after SAH.(3) The sham+BGN lentivirus group had statistically lower BGN mRNA expression than the sham+control lentivirus group (P＜0.05).(4) As compared with those in the SAH+control lentivirus group,both scores of modified Garcia scale and beam balance tests were significantly higher in SAH+BGN lentivirus group (6.125±1.246 vs.13.000±1.309;1.125±1.126 vs.2.875±0.835),and neural apoptosis ratio and ratio of p-NF-κB/NF-κB were significantly lower in the SAH+BGN lentivirus group (51.950％±11.166％ vs.31.938％±7.705％;1.161±0.156 vs.0.886±0.142,P＜0.05).Conclusion Inhibition of BGN can effectively reduce neuronal apoptosis in mice with EBI after SAH,and attenuate neurological deficits.

Objective To explore the altered expression of circular RNA (circRNA) and mRNA in the mouse cortex in the early phase of subarachnoid hemorrhage (SAH) and possible biological functions of the circRNA in early brain injury (EBI).Methods A total of 18 C57BL/6J male mice were randomly divided into a sham and a SAH group (n=9).SAH models were prepared by endovascular perforation.Total RNAs of brain samples were extracted to construct the cDNA library 24 h after operation.RNA-sequencing (RNA-seq) was carried out by HiSeqTM 2500 User Guide and followed by RT-qPCR for confirmation.Reads were aligned to the mouse transcriptome to obtain expression profiles ofcircRNA and mRNA.Bioinformatic study included GO analysis,KEGG pathway analysis and forecast of targeted miRNA of circRNA.Results A total of 26 circRNA (6 up-regulated and 20 down-regulated) and 804 mRNA (396 up-regulated and 408 down-regulated) were significantly changed.These altered mRNA were mainly related to regulation of neuronal synaptic plasticity,inflammatory and immune response.Bioinformatics showed that some significantly altered circRNA contained binding sites for many miRNA.The RT-qPCR analysis of 4 randomly selected circRNA (circFoxj3,circSetbp1,circArpp21 and circ2010111 I01Rik) confirmed the accuracy of RNA-seq.Conclusions SAH alters the expression of circRNA in mouse cortex and the differentially expressed circRNA may be involved in regulation of EBI following SAH,promising a potential therapeutic target for the diagnosis,treatment and prognosis of SAH.

Objective To study the effects of main ingredients from microemulsion extract of Compound Longqincao in vitro on anti-influenza virus H1N1 activity; To analyze effects of main ingredients from microemulsion extract on influenza virus inhibition rate.Methods Uniform design was used to conduct the experiment. MTT method was used to detect the effect rate (ER) of anti-influenza virus H1N1 on A549 cells. Setting ER as the index, Minitab17 software was used to establish mathematical model to come up with regression equations of all factors. The effects of ingredients on ER were analyzed and the efficient composition ratio of the optimum anti-influenza virus H1N1 was chosen.ResultsIn the compound compatibility, baicalin showed the most obvious antivirus activity, and licorice glycosides had certain inhibition effects on pathological changes of cells. Five ingredients had coordinative or controlled relation with ER. When per milliliter liquids containing licorice glycosides, baicalin, leucine, aspartic acid, glutamic acid was 13.94μg, 49.44μg, 0.23 mg, 1.25 mg, and 2.50 mg, ER was the best. ER was 85.34%±4.72% after verification.ConclusionThe optimized combination of main ingredients from microemulsion extract of Compound Longqincaocan better play a role in anti-influenza virus H1N1.

Objective: To analyze the clinical features and outcomes of cardiac resynchronization therapy (CRT) in patients with dilated-phase hypertrophic cardiomyopathy (DHCM). Methods: A total of 16 DHCM patients received CRT in our hospital from 2007-03 to 2016-01 were retrospectively studied to analyze their clinical features and outcomes. Results: There were 12 male and 4 female patients at the mean age of (53.3±13.5) years. Pre-operative QRS duration of ECG was (158.7±32.2) ms, left ventricular ejection fraction (LVEF) was (33.6±6.3) %, the patient with NYHA class I, II, III and IV were 1, 5, 8 and 2 respectively. 13 patients received new CRT device, 3 received upgraded device and 8 (50%) combining atrial fibrillation (AF). The patients were followed-up for (2.56±2.13) years, 5 of them died including 3 of heart failure, 1 of sudden death and 1 of stroke. At 6 months follow-up time, 7 patients had the response to CRT which was defined by the improvement of NYHA class≥1 and the absolute elevation of LVEF≥5%; NYHA class improved from (2.69±0.79) to (2.38±0.89), P=0.02; LVEF increased from (33.6±6.3) % to (40.03±9.83) %, P=0.01. Conclusion: DHCM patients with CRT indication had the higher incidence to suffer from AF, those were more in patients with traditional pacemaker or ICD upgrading. DHCM patients with CRT had the poor general prognosis, while there was still certain proportion of patients had the response to CRT.

Ideal blood glucose control requires accurate insulin injections under the guidance of frequent glucose monitoring.Artificial pancreas (AP),the closed-loop control system can adjust the input amount of insulin automatically with the body's blood glucose levels.The AP allows diabetics to control blood glucose ideal,then get the benefit of prevention of complications and bring convenience and safety in clinical application.Accuracy is the key issue of the AP.To improve the accuracy of such a system need to improve the detection accuracy and reliability,increase speed and accuracy of the output control,and improve the accuracy of the system regulation model.

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.

Aim To investigate the regulatory effects of baicalin on apoptosis induced by virus H1 N1 in hu-man pulmonary carcinoma cell A549 . Methods The chips were used to screen the RNA samples in virus-in-fected A549 cells. Differentially expressed genes were selected in the pathway of apoptosis. The mRNA ex-pressions of caspase-3 and -8 were verified by Real-Time PCR. Results With the DNA microarray, the functions of differentially expressed genes involved in apoptosis biological pathways were analyzed by Kyoto Encyclopedia of Genes and Genomes ( KEGG ) Path-way databases. caspase-3, -7, -8, -10, TRAIL, MYD88 , IL1 A and IL1 B were up-regulated in virus-in-fected group. Oseltamivir could down-regulate gene ex-pressions of caspase-3 ,-4 and-8 . High-dose of baica-lin could down-regulate gene expressions of caspase-3 ,-4,-6 and -8. Except gene expressions of above, low-dose of baicalin could also down-regulate gene expres-sions of IL1RAP and Cn. Real-Time PCR experiments showed that baicalin could significantly decrease mR-NA expression of caspase-3 , -4 , -6 , -8 , IL1 RAP and Cn ( P < 0. 01 ) , compared with the virus-infected group. The results also figured that the interference ef-ficacy of low-dose baicalin was better than that of high-doses. As expected, real-time PCR data were in good agreement with the microarray assay. Conclusions Baicalin can be detected in their suppression effect of caspase-3,-4,-6, and -8 mRNA expression, so it re-sists against the apoptosis to fight against influenza vi-rus in vitro.

Objective To investigate the effects of Tangshen Formula on liver oxidative stress in diabetic rats, and their mechanisms thereof.Methods Rat model of type 2 diabetes mellitus and nonalcoholic fatty liver disease was estab?lished by intraperitoneally injecting small dose of chain urea with cephalosporins (STZ) and feeding high fat fodder . The model rats were randomly divided into control group, model group, Tangshen Formula group and metformin group.The se?rum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglyceride (TG), cholesterol (CHO), low density lipoprotein-cholesterol(LDL-C), and superoxide dismutase (SOD), catalase(CAT), vitamin E(VE), nitric oxide (NO) and nitric oxide synthase (NOS) in liver were compared between four groups. Changes of pathological morphology were ob?served under light microscopy. Results There were significantly decrease in serum levels of ALT, AST, TG, CHO, LDL-C in Tangshen Formula group and metformin group compared with those of model group. There were significantly higher levels of SOD, CAT and VE, and lower levels of NO and NOS of liver homogenate in Tangshen Formula group than those of model group. There were higher levels of SOD and CAT, and lower levels of NO and NOS in liver homogenate in metformin group than those of model group. HE staining showed that liver fatty degeneration was significantly reduced in metformin group and Tangshen Formula group compared with that of model group. Conclusion The fatty liver in type 2 diabetic rats is signifi?cantly improved by Tangshen Formula treatment, which is probably associated with the regulation of the response level to oxi?dative stress in liver.

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.

Objective To establish standard sites for bur holes that maintain constant anatomical relationships with the skull base and neural structures and can serve as the basal aspect of supratentorial temporooceipital craniotomies such as subtemporal transpetrosalridge approach.Methods To determine cranial-cerebral relationships,the authors delimited 10 adult cadaveric skulls anteriorly and posteriorly the external projection of the petrous bone and the midbrain by CT and Titanium nail.Then bur holes in adult cadaveric skulls were created (kl,the first bur hole,located anterior to the auricle of the ear; k2,the second bur hole,whose base was located 1 cm above the interface of the parietomastoid and squamous sutures; k3,the third bur hole whose base was located 1cm above the asterion).Three bur holes were made on each of the skulls (20 cerebral hemispheres).The author then introduced plastic catheters through the bur holes to evaluate pertinent cranial and neural landmarks.Results The first bur hole appeared to have a particular anatomical relationship with the anterior aspect of the petrous portion of the temporal bone and the most anterior aspect of the midbrain.The second bur hole had a particular relationship with the posterior border of the petrous portion of the temporal bone and with the posterior aspect of the midbrain.The third bur hole was mostly supratentorial and particularly related to the preoccipital notch.Conclusions The middle fossa floor is located anterior to the site of the kl,and the superior surface of the tentorium is posterior to k2.Together with k3,these bur holes can be considered standards for temporooccipital craniotomies such as subtemporal transpetrosalridge approach.