Objective To develop a new method of immune isolation for transplanting pig islets to rat to observe the effects on diabetic rats.Method Diabetes of rat was induced by streptozotocin.Pancreas of pig was digested with type V collagenase.The pig islets were purified by density gradient centrifugation.Insulin stimulation index of the purified pig islets was evaluated.Liver of the 18 diabetic rats and 6 normal rats was exposed by operation.Capsule at one edge of the liver lobe was cut apart and separated from liver parenchyma on both sides of the lobe.An artificial cyst was constructed by a cellulose ester (CE) dialysis bag with a ball of hollow fibers in the bag,and each end of the bag was closed by a thread.Each end of the bag was stuffed into the subcapsule of each side of the liver lobe.The thread at each end of the bag was passed through a hole of the capsule at other edge of the lobe and linked to each other around the lobe to hold the artificial cyst on the liver parenchyma.Middle part of the cyst (with the ball of hollow fibers in) was put between the cuts of capsule and under the skin.The collagen solution (pH 7.4) with pig islets (4000 IEQ) was injected into the subcapsule artificial cyst of 12 diabetic rats as experiment group.The collagen solution without pig islets was injected into the subcapsule artificial cyst of 6 diabetic rats and 6 normal rats as diabetes control group and normal control group respectively.The cut was sutured.The collagen solution became a collagen gel in the artificial cysts.At 8th week of the initial transplantation,the solution with pig islets (2000 IEQ) was injected percutaneously again into the subcapsule artificial cyst of each rat of the experiment group.Rats of the three groups were raised as usual,and the levels of fasting blood glucose were measured regularly.Result The insulin stimulation index of the purified pig islets was 2.2 ± 0.2.At 1st to 8th week of the initial transplantation,the levels of fasting blood glucose in the experiment group at every time point were lower significantly than those in the diabetes control group (P＜0.01).At 13th week of the initial transplantation,there was no significant fibroplasia around the subcasule artificial cysts in the rats by visual observation.Conclusion CE dialysis bag in the liver capsule at 13th week did not show significant stimulation to the surrounding tissues.Pig islets isolated in the subcapsule artificial cyst of rats might live normally and perform hypoglycemic action.Pig islets could be percutaneously injected into the subcapsule artificial cyst.

Objective Investigate the clinical efficacy of treating retreated smear-positive tuberculosis patients with conventional anti-tuberculosis chemotherapy combined with levofloxacin. Methods Divide 60 retreated smear-positive tuberculosis patients registered in our hospital between October 2012 and October 2014 into two groups using random number table method: levofloxacin group and conventional treatment group. Each group contains 30 patients that were treated for 9 months , and the efficacy the both treatment methods were compared. Results The sputum conversion rate in the levofloxacin group is 93.33% and 96.67% after treating for 6 months and 9 months , respectively; which are substantial higher than that of the conventional treatment group with sputum conversion rate of 70% and 73.33% after 6 months and 9 months of treatment , respectively (P < 0.05). The absorption rate of the levofloxacin group is significantly higher than that of the conventional treatment group (P < 0.05). The cavity improvement rate of the levofloxacin group is 73.33%, which is significantly higher than that of the conventional treatment group (33.33%) (P < 0.05). The clinical efficacy of the LVFX group is substantially higher than the routine treatment group (P < 0.05). The overall effectiveness of LVFX group reached 100% , which is significantly higher than that of the conventional treatment group (86.67%) (P < 0.05). Conclusions Levofloxacin combined with conventional anti-tuberculosis chemotherapy can effectively improve the clinical efficacy in the treatment of tuberculosis.

Objectives To evaluate the performances of transcranial Doppler (TCD) and vessel-encoded arterial spin labeling MRI perfusion imaging (VE-ASL) in the evaluation of collateral circulation of cerebral arteries. Methods Thirty patients with unilateral ICA or MCA stenosis diagnosed by TCD and carotid duplex ultrasound and confirmed by MRA underwent VE-ASL. Peak systolic velocity (PSV) of bilateral MCA in the same depth, distal to the stenosis site, was recorded. Cerebral blood flow (CBF) in bilateral MCA territories was measured by VE-ASL. The detection rates of collater-al flow using TCD and VE-ASL were compared. Results TCD revealed that PSV in the ipsilateral and contralateral MCA were (31.6 ± 10.5) cm/s and (83.1 ± 9.2) cm/s, respectively. VE_ASL revealed that CBF in the ipsilateral and contralateral MCA territory were (22.5±9.8) mL·min-1·100g-1 and (31.7±8.3) mL·min-1·100g-1, respectively. The PSV ratio of the ipsi-lateral/contralateral MCA was significantly lower than the CBF ratio of the ipsilateral/contralateral MCA(0.37 ± 0.173 vs. 0.66±0.141, P=0.001). The detection rates of collateral flow using TCD were lower than those using VE-ASL. The detec-tion rate was 26.7% vs. 70% on TCD vs. VE-ASL in anterior collateral circulation (P=0.001), was 16.7% vs. 60% (P<0.001) on TCD vs. VE-ASL in posterior collateral circulation. The total display rate of collateral flow was 36.7%vs. 86.7%on TCD vs. VE-ASL (P<0.001). Conclusions TCD is inferior to VE-ASL in evaluating the collateral circulation because of the limitations including the anatomical variations of the circle of Willis and formation of leptomenigeal anastomoses.

Objective To evaluate the diagnostic value of the early rheumatoid arthritis (RA) classification criteria for early RA and to assess the clinical efficacy of knee arthroscopic synovectomy for RA.Methods A total of 62 patients who had arthritic complaints with disease duration less than one year were treated with knee arthroscopic synovectomy.Patients were diagnosed as RA confirmed by changes of synoviomorphous under arthroscopy,synovial biopsy,immunological biochemical laboratory and MRI,and compared with the preoperative ERA diagnosis.The efficacy was assessed at the third months and more than 12 months after operation,including Health Assessment Questionnaire (HAQ),Lysholm score,laboratory parameters of erythrocyte sedi-mentation rate (ESR) and C-reactive protein (CRP).Results The sensitivity and specificity in the early RA classification criteria for RA diagnosis were 81.58％ and 91.67％.38 patients diagnosed as RA and the remaining 24 patients were significantly improved of HAQ,Lysholm score,ESR,CRP compared with preoperation,showed statistical difference (P ＜ 0.01).There were still statistical differences between the follow-up results of the group of patients with RA in different period after operation (P ＜ 0.01).Conclusion The early RA classification criteria is worthy of clinical application,because it has the advantages of simple,practical,and is beneficial to early diagnosis of RA.The early RA patients should be treated with knee arthroscopic synovectomy as far as possible.It can improve not only the knee function,but also the general symptoms,and it can interrupt the RA pathological process,effective maintain curative effect after operation.

Objective To analyze the composing experience of Menghe physician Ma Peizhi for cough by TCM inheritance support system. Methods The prescriptions for cough of Ma Peizhi were collected, frequency and association of drugs were analyzed by using data mining methods such as revised mutual information, complex system entropy cluster and unsupervised hierarchical cluster. Results Based on the analysis of 57 prescriptions, the frequency of each herb and association rules among the herbs were computed, 18 core combinations and 9 new prescriptions were mined from the database. Conclusion Menghe physicians Ma Peizhi is well experienced in expelling wind and opening the inhibited lung, dissolving phlegm and relieving cough. TCM inheritance support system can be used to analyze clinical experience of old TCM doctor.

Objective To analyze the principle of the drug use ofMenghe PhysiciansMa-Peizhi by using the Traditional Chinese Medicine(TCM)inheritance support system.Methods The prescriptions for the commonly encountered diseases that used byMa-Peizhi were collected to build a database, and analyze by the unsupervised data mining methods, such as apriori algorithm, entropy clustering complex systems, from which we could get the frequency of the drugs, the association rules between drugs, the core drug combinations, and so on.Results Based on the analysis of 745 prescriptions, the most frequently used drugs were tuckahoe, chiretta, paenoiae alba, dried orangepeel and dioscoreae. The core drug combinations were “radix rehmanniae recen- salivia chinensis-ophiopogon root”, “teasel root-viscum album-achyranthes”, “menispermaceae-heracleum hemsleyanum michaux-gentiana macrophylla”, and “mulberry leaf-periostracum cicadae-the root of balloon flower”. The new prescriptions were “mulberry leaf-viter rotundifolia-batryticated silkworm-periostracum cicadae-the root of balloon flower”, “teasel root-viscum album- achyranthes- ramulus mori- periplocae”, and so on.ConclusionMenghe PhysiciansMa-Peizhi was well experienced in treating the commonly encountered diseases by agile diagnosis and treatment, and addition or subtraction of changes based on the classical prescriptions.

Abstract: Objective To explore the prevalence and bacterial strains of Francisella tularensis (F. tularensis) in wild rodents in Changbai Mountain area of China, and to further understand the epidemiological characteristics of F. tularensis infections in this area. Methods Wild rodents were captured from forest and forest-edge farmland from Kuandian County and Jianshi Forest District in the Changbai Mountain area, 2012-2014. DNA was extracted from the spleen tissues of the rodents, and the fopA gene of F. tularensis in wild rodents was detected using nested PCR. The infection rates were calculated for different areas and rat species. The bacterial subspecies of positive samples were identified using type-specific primers (C1/C4), and sequencing and comparative analysis were performed. Results A total of 133 wild rodents belonging to 6 rat species were captured. Among them, eight samples from three rat species (Apodemus agrarius, Apodemus peninsulae, Tscherskia triton ) were detected positive, with the overall positive rate of 6.01%. The positive rates of F. tularensis of Ji'an and Kuandian were 7.46% and 4.54%, respectively, and there was no difference in positive rates for different regions (χ2=0.117, P=0.732) and different rat species (χ2=0.641, P=0.986). The subspecies analysis showed that the detected 8 trains of F.tularensisall belonged to F.tularensis type B (F.subspecies subsp. holarctica). Genetic evolution analysis was performed on the fopA gene sequences of three positive samples (JA56, JA33, and JA38), which clustered together with Russia strains(CP009694.1, CP044004.1) and China strains (HM371344.1, HM371343.1) F.tularensis type B, with sequence similarities ranging from 99.21% to 99.47%. Conclusions Infection of F.tularensis subsp. holarctica existed in wild rodents in Changbai Mountain area of China, which suggests the existence of F.tularensis infection risks in this area.

Objective To assess the inter-scanner reliability and reproducibility of 3D whole-brain pseudo-continuous arterial spin labeling (pCASL) of posterior circulation territory in healthy adults.Methods Twelve healthy subjects were scanned twice on two different 3.0 T MR scanners with 3D wholebrain pCASL technique.Intervals between tests were among 10-15 days.The ASL data with two post labeling delay time (PLD) of 1.5 and 2.5 seconds were acquired during every scanning.The cerebral blood flow (CBF) values of posterior circulation territory including cerebellum,midbrain,pons,medulla oblongata,thalamus and occipital lobe were extracted based on SPM technique for comparison.The interscanner reliability and reproducibility were evaluated with the intraclass correlation coefficient (ICC) and Bland-Altman plot.Results The CBF values of posterior circulation territory were 36-47 ml · min-1 ·100 g-1 (PLD =1.5 s) and 36-46 ml · min-1 · 100 g-1 (PLD =2.5 s).Comparing the data between the two tests,the ICC was 0.851 at PLD =1.5 s and 0.914 at PLD =2.5 s.The Bland-Altman showed the reproducibility at PLD =2.5 s was better than that at PLD =1.5 s.Conclusions The 3D whole-brain pCASL technique is available for measuring the CBF at posterior circulation territory with high reliability and reproducibility especially using longer PLD.It can be used for MR multicenter study on blood flow of posterior circulation territory.

OBJECTIVETo investigate the effects of different concentrations of lipopolysaccharide (LPS), tumor necrosis factor α (TNFα), interleukin-6 (IL-6), dexamethasone (Dex), and insulin on the mRNA and protein expressions of GPR54 in the MCF7 cell line in vitro.

METHODSMCF7 breasr cancer cells were cultured and treated with different concentrations of LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L). Those treated with culture fluid only served as controls. The mRNA and protein expressions of GPR54 were measured by real-time PCR and Western blot, respectively, after 6, 24, 48, and 72 hours of treatment.

RESULTSCompared with the blank con- trol, LPS (10 and 20 µg/ml), TNFα (20 and 100 ng/ml), IL-6 (10 and 20 ng/ml), Dex (10(-6) and 10(-7) mol/L), and insulin (0.01 and 0.1 IU/L) significantly increased the expressions of GPR54 mRNA (P < 0.05) and protein (P < 0.05).

CONCLUSIONLPS, TNFα, IL-6, Dex, and insulin evidently increase the expression of GPR54 in the MCF7 cell line, indicating their influence on the function of gonads by regulating the GPR54 level.

Blotting, Western ; Dexamethasone ; administration & dosage ; pharmacology ; Glucocorticoids ; administration & dosage ; pharmacology ; Gonads ; drug effects ; metabolism ; Humans ; Hypoglycemic Agents ; administration & dosage ; pharmacology ; Insulin ; administration & dosage ; pharmacology ; Interleukin-6 ; administration & dosage ; pharmacology ; Lipopolysaccharides ; administration & dosage ; pharmacology ; MCF-7 Cells ; RNA, Messenger ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptors, G-Protein-Coupled ; drug effects ; genetics ; metabolism ; Receptors, Kisspeptin-1 ; Time Factors ; Tumor Necrosis Factor-alpha ; administration & dosage ; pharmacology

Objective To observe the clinical efficacy of deproteinized calf blood extractive injection combined with hyperbaric oxygen in patients with severe traumatic brain injury.Methods 74 patients with severe traumatic brain injury from January 2013 to March 2015 in Wenzhou hospital of Chinese traditional medicine were randomly divided into observation group and control group, 37 cases in each group.The control group received hyperbaric oxygen on the basis of conventional therapy, the observation group received deproteinized calf blood extractive injection on the basis of control group.The Glasgow coma scale ( GCS) , Barthel, hs-CRP, TNF-α, IL-6 and prognosis were compared between two groups.Results The GCS score and Barthel index scores post-treatment in observation group were (13.67 ±1.73),(65.73 ±4.02) points, which were higher than (9.66 ±1.24), (50.69 ± 3.76) points in control group, and the difference was significant (P<0.05).The serum hs-CRP, TNF-α, IL-6 post-treatment in observation group were (4.55 ±0.76)mg/L,(1.21 ±0.05)μg/L,(0.21 ±0.01)μg/L, which were better than those of control group (6.43 ±1.01)mg/L,(1.36 ±0.06)μg/L,(0.28 ±0.02)μg/L (P<0.05).The rate of favorable prognosis in observation group was 48.65%, which was higher than that of control group (P<0.05).Conclusion The deproteinized calf blood extractive injection combined with hyperbaric oxygen has the exact efficacy, which was better than hyperbaric oxygen alone in the treatment of severe traumatic brain injury.