Objective: To optimize the preparation technology for total glucosides from Paeonia lactiflora (TGP) liposome and study on its in vitro release. Methods: Liposomes were prepared by reverse-phase evaporation technique. Taking the entrapment efficiency (EE) as indexes, the preparation of liposome was optimized using Box-Behnken design-response surface method (BBD-RSM), and the in vitro release was studied. Results: The optimal formulation was cholesterol-lecithin (1:2.7), TGP-lipid (1:29), and hydrated medium of 5.1 mL. The average EE were 73.0% and the 12 h in vitro accumulative release rate was 42.18%. Conclusion: The optimized process is rational and feasible, and the TGP liposome has the higher EE and good stability with better sustained release in vitro.

OBJECTIVE: To identify Angelica sinensis and its adulterants by ISSR markers. METHODS: Genome polymorphism of 26 A. sinensis and its adulterants were analyzed by ISSR-PCR. The genetic similarity coefficients and genetic relationships were calculated by NTSYS-pc version 2.10. The dendrogram was constructed based on UPGMA clustering. RESULTS: Ten out of 100 ISSR primers are screened out and 96 bands were amplified, among which 85 were polymorphic fragments (percentage of polymorphic bands was 88.54%). The length of bands ranged from 200 to 2100 bp. One specific band was amplified by the primer UBC848.A. sinensis and its adulterants could be discriminated by primer UBC848 and UBC834. Results of UPGMA dendrogram indicated that A. sinensis from nine different places clustered together and related to A. acutiloba. The other species were clustered far away from A. sinensis. Three commercially crude materials purchased in the market were adulterants. CONCLUSION: The specific primers were found to identify A. sinensis by ISSR method. There is genetic diversity among A. sinensis from different production areas. The results provide molecular evidence for identification of A. sinensis and its adulterants and technical support of building up the stable quality evaluation system of A. sinensis.

OBJECTIVE: To explore the relationship between serum cystatin-C (Cys-c) levels and vibrating perception threshold (VPT) in patients with Type 2 diabetes mellitus (T2DM).
 METHODS: According to the symptoms, signs and results of lab examination, a total of 352 patients with T2DM were divided into a diabetic peripheral neuropathy group (DPN group, n=107) and a non-diabetic peripheral neuropathy group (NDPN group, n=245). Serum Cys-c levels were measured by radioimmunoassay method. The relationship between serum Cys-c levels and VPT, estimated glomerular filtration rate (eGFR), urinary albumin-to-creatinine ratio (ACR), glucose and blood pressure and other parameters were also analyzed by correlation and multiple regression analysis. All T2DM patients were divided into a high Cys-c levels group (n=89) and a low Cys-c levels group (n=263) according to the upper quartile of Cys-c, and the incidence of DPN and VPT levels in each group were compared. Risk factors of DPN in T2DM patients were analyzed by binary logistic regression analysis and receiver operating characteristic (ROC) curve was used to identify the optimal cutoff of serum Cys-c levels for predicting DPN in patients with T2DM.
 RESULTS: Serum Cys-c levels were significantly higher in the DPN group than that in the NDPN group [(1.04±0.43) vs (0.80±0.25) mg/L, P<0.01]. Correlation analysis showed that serum Cys-c levels were positively correlated with age, body mass index (BMI), serum creatinine (SCr), blood urea nitrogen (BUN), ACR, VPT, pulse pressure (PP), white blood cell( WBC) , red cell distribution width (RDW), hemoglobin A1c (HbA1c) and fasting blood glucose (FBG) (r=0.410, 0.115, 0.613, 0.433, 0.291, 0.300, 0.156, 0.129, 0.282, 0.314, 0.236, respectively, P<0.05 or P<0.01); and negatively correlated with diastolic blood pressure (DBP), eGFR and indirect bilirubin (IBIL) (r=-0.135, -0.647, -0.114, respectively, P<0.05 or P<0.01). Serum Cys-c levels in T2DM patients were positively correlated with VPT after adjusting for gender, age, BMI, ACR and eGFR (r=0.235, P<0.01). Multiple regression analysis revealed that VPT, age, SCr, eGFR, PP, ACR and HbA1C were independent related factors affecting serum Cys-c levels in T2DM patients. Compared with those in the low Cys-c levels group, the prevalence rate and VPT value was increased in the high Cys-c levels group (all P<0.01). Binary logistic regression analysis found that age, Cys-c and HbA1C were independent risk factors for predicting DPN in T2DM patients (all P<0.01). ROC curve analysis revealed that the optimal cutoff of Cys-c to predict DPN in T2DM patients was 0.996 mg/L, the sensitivity was 43.9%, the specificity was 83.7%, and the area under curve was 0.663.
 CONCLUSION Serum Cys-c levels are well correlated with VPT in patients with T2DM. When the serum Cys-c levels>0.996 mg/L, the predicts have high risk of DPN in T2DM patients, which might be related to diabetic nephropathy, oxidative stress and inflammatory reaction induced by advanced age, hyperglycemia and hypertension.
Cystatin C ; blood ; Diabetes Mellitus, Type 2 ; blood ; Diabetic Neuropathies ; blood ; Humans ; Incidence ; Perception ; physiology ; Prevalence ; Risk Factors ; Vibration

Objective: To explore the position change of the setup reference markers on the sternum due to the change of hands grip position. Methods: Six hands grip positions were simulated for 8 patients and the set-up reference points marked on the sternum skin were compared while the hands grasped different positions of the hands. Results: The setup markers on the sternum were deviated by all five hands grip positions. The maximum deviated distance was 14mm. Conclusion: The change of the hands grip position can affect the position of setup reference markers on the sternum of the patients and thus the isocenter of plan may be aligned to the wrong area.

Objective To establish a rat model of associating liver partition with portal vein ligation for staged hepatectomy (ALPPS) and evaluate the liver function after surgery.Methods Fifty male SD rats were randomly divided into two groups: experimental group (ALPPS group) and control group (PVL group).Selective portal vein ligation in PVL group was performed on the caudal lobe, left lateral and left median lobes, while the right lobe, the right median lobe was preserved to regenerate.ALLPS group was treated in the same way as PVL group, but also underwent liver partition in situ.After surgery, 5 rats were sacrificed on day 1, 3,7, 10 and 14 in each group to observe the weight of body and the right median lobe,respectively.The venous blood and liver tissue were obtained for testing alanine aminotransferase (ALT),aspartate aminotransferase (AST), serum albumin (ALB), total bilirubin (TBil) and observing the histological changes in liver injury after surgery.Results After surgery, the body weight decreased progressively, but then increased in both groups.Since the first day after surgery, the body weight began to decrease,reached the lowest value on 3 d, and then on day 7 the body weight in PVL group returned to preoperative levels.However, the body weight was still lower than that before surgery [(3.7 ± 2.7) ％ vs (-3.0 ± 1.9)％, P＜0.05].On day 3, 7, 10 and 14, the hepatic regeneration rate (HRR) of the fight median lobe in ALPPS group was obviously higher than that in PVL group (P ＜ 0.05).On day 1, the serum ALT and AST levels in two groups were elevated dramatically and then gradually decreased, which in ALPPS group were significantly higher than those in PVL group (P ＜ 0.05).There were no significant differences at other time points.On day 1 and 3, the serum ALB in ALPPS group was obviously lower than that in PVL group [(25.4±1.7)g/Lvs (31.4±1.5)g/L, P＜0.05;(25.0±2.0)g/Lvs (31.8±1.5)g/L, P＜ 0.05], respectively.More focal necrosis of liver were observed in ALPPS group on day 1, which were more than those in PVL group.Conclusions This method could successfully establish a ALPPS rat model and proved that ALPPS could induce accelerated hepatic regeneration and more severe early hepatocyte injury compared with PVL.This ALPPS experimental model provides a basis for further research on ALPPS, especially for clarifying the mechanisms of liver regeneration and tumor recurrence, and exploring the reasons for various ALPPS related complications, which play a significant role in its clinical application.

Objective To determine the tumor clonal origin of diffuse hepatocellular carcinoma (D-HCC) and clinical outcome after liver transplantation. Methods Fourteen D-HCC patients who underwent liver transplantation in the Tianjin First Central Hospital between August 2004 to November 2012 were evaluated retrospectively. Their clinical characteristics and data of disease-free survival postoperatively were gathered. Twelve loss of heterozygosity sites (LOH) on high frequency microsatellite were selected. The tumor clonal origin of D-HCC was assessed by the means of PCR-SSCP. Results Of 14 D-HCC patients, intrahepatic metastasis (IM) occurred in 11 cases (11/14), and IM with multicentric occurrence (MO) occurred in 3 cases (3/14). The disease-free survival time was 4.5 to 37.4 months, and the median time was 8.7 months. Conclusion The most frequent tumor clonal origin of D-HCC was IM. Liver transplantation may be the effective treatment for D-HCC.

Objective To explore the influence of intra-abdominal fat (IAF) accumulation on ambulatory blood pressure (ABP). Methods Four hundred and forty-eight hospitalized patients with cardiovascular and metabolic diseases, including 235 males and 213 females, aged 20-85 (55±12) years old, were enrolled in this study. The abdominal fatty thickness was measured with B-mode ultrasound. Simultaneously the patient received 24hABP monitoring. The patients were divided into excessive IAF group, normal group, and quintile subgroups according to the cut-off points (male 38.5mm, female 34.7mm) for observing the difference in indexes of ambulatory blood pressure between different groups/subgroups. Results The 24h mean systolic BP (24hMSBP), 24h mean BP (24hMBP), 24h mean diastolic BP (24hMDBP), daytime MSBP (dMSBP), daytime MBP (dMBP), daytime MDBP (dMDBP), nighttime MSBP (nMSBP), nighttime MBP (nMBP), night MDBP (nMDBP) were significantly higher in the excessive IAF group than in the normal group (P < 0.05). However, the elevation of ABP was not a simple gradual linear elevation corresponding to a gradual increase in IAF thickness, but manifested an approximately U-shaped trend. Pearson correlation analysis showed IAF thickness was positively correlated with all the indexes of 24hABP, while partial correlation analysis after adjustment for age and sex showed IAF thickness was positively correlated with 24hSBP, 24hMBP, dMBP and nMBP. Conclusion The relationship between IAF thickness and ABP presents a U-shaped curve, which may provide an enlightenment in controlling blood pressure by weight loss in clinical work.

This paper aims to establish a new method of calibration and positioning in quantitative analysis of multicomponents by single marker (QAMS), using Shuanghuanglian oral liquid as the research object. Establishing relative correction factors with reference chlorogenic acid to other 11 active components (neochlorogenic acid, cryptochlorogenic acid, cafferic acid, forsythoside A, scutellarin, isochlorogenic acid B, isochlorogenic acid A, isochlorogenic acid C, baicalin and phillyrin wogonoside) in Shuanghuanglian oral liquid by 3 correction methods (multipoint correction, slope correction and quantitative factor correction). At the same time chromatographic peak was positioned by linear regression method. Only one standard uas used to determine the content of 12 components in Shuanghuanglian oral liquid, in stead of needing too many reference substance in quality control. The results showed that within the linear ranges, no significant differences were found in the quantitative results of 12 active constituents in 3 batches of Shuanghuanglian oral liquid determined by 3 correction methods and external standard method (ESM) or standard curve method (SCM). And this method is simpler and quicker than literature methods. The results were accurate and reliable, and had good reproducibility. While the positioning chromatographic peaks by linear regression method was more accurate than relative retention time in literature. The slope and the quantitative factor correction controlling the quality of Chinese traditional medicine is feasible and accurate.
Apigenin ; analysis ; Calibration ; Chlorogenic Acid ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Flavanones ; analysis ; Flavonoids ; analysis ; Glucosides ; analysis ; Glucuronates ; analysis ; Glycosides ; analysis ; Reproducibility of Results

OBJECTIVEMuscle segment homeobox gene (MSX)1 has been proposed as a gene in which mutations may contribute to nonsyndromic cleft lip with or without cleft palate (NSCL/P). To study MSX1 polymorphisms in NSCL/ P by means of polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP), and investigate the association of MSX1 exons 1 polymorphisms with NSCL/P.

METHODSDNA were extracted from blood samples from NSCL/P and unrelated normal subjects. Genome DNA from peripheral leukocyte with these blood samples were extracted, which was used as template to amplify desired gene fragment of MSX1 exons 1 by means of polymerase chain reaction (PCR). The PCR products were examined by single-strand conformation polymorphism (SSCP). The MSX1 exons 1 polymorphisms were examined by sequencing if mutations were found.

RESULTSMSX1 genes of exon 1 mutation was not been found in the NSCL/P and unrelated normal subjects by SSCP.

CONCLUSIONNo correlation between MSX1 exon 1 and NSCL/P was found. MSX1 exon 1 may not be a key gene (susceptibility gene) in NSCL/P.

The current appointed teaching material of Science of Acupuncture and Moxibustion holds that there is no difference among the needling sensation, arrival of qi and needling response. However, the author has a different understanding. Therefore, Neijing (Internal Classic), its annotation, exposition and understandings of ancient and modern famous experts are cited to analyze their meanings. And the result indicates that the needling sensation is subjective feelings and perceived responses of doctors and patients. Arrival of qi is the healing process of the organ through activating the anti-pathogenic qi to expel the pathogens. The needling response is the final aim of acupuncture therapy. Thus, the meaning of needling sensation, arrival of qi, and needling response are different. And an accurate understanding can better guide acupuncture treatment.
Acupuncture Therapy ; instrumentation ; methods ; Humans ; Moxibustion ; instrumentation ; methods ; Needles ; Qi