Soxhlet extraction is a common method of sample preparation. However, there has been no discussion about the efficiency of Soxhlet extraction from different batches and the factors that cause content fluctuation. In this study, Panax ginseng was selected as a model sample. Soxhlet extraction by means of a water bath, which has always been neglected, was identified as a novel key factor in the poor repeat-ability in different batches of Soxhlet extraction, as it can affect the siphon times and reflux time, which have been positively correlated with the ginsenoside contents. By substituting round bottom flasks in the same column, the relative standard deviation of the most fluctuated compound, ginsenoside Rb1, was decreased from 24.6% to 5.02%. Scanning electron microscopy analysis confirmed that the breakdown of the surface of the ginseng powder in the Soxhlet extraction led to a better dissolution of ginsenosides, indicating that chloroform may promote the extraction of ginsenosides by disrupting the cell structure. Moreover, 70% methanol was regarded as the better solvent for extracting the ginsenosides. Overall, this work offers a practical and effective protocol for improving the accuracy and repeatability of Soxhlet extraction methodology for ginsenosides and other analytes.

The aim of study was to investigate the killing effect of double suicide gene system mediated by retroviral vector on K562 cells in vivo and ex vivo. CDglyTK gene was transfected into PA317 cells by using lipofectamine. K562 cells were infected with viral supernatant. K562/CDglyTK cells were treated with 5-fluorocytosine (5-FC) and/or ganciclovir (GCV). Mice were randomly divided into three groups: tumor formation, tumor inhibition and tumor therapy. Each mouse was implanted with K562/CDglyTK cells or K562 cells. The results indicated that the killing effect of 5-FC in combination with GCV on K562/CDglyTK was more significant than using 5-FC or GCV alone. In vivo study showed that after being injected subcutaneously with K562 cells and K562/CDglyTK cells, there was not obvious difference in tumor formation rate of mice, 5-FC + GCV could suppress tumor formation of the K562/CDglyTK cells. After being treated with 5-FC and GCV, the median tumor volume of mice implanted with K562/CDglyTK cells decreased obviously, compared with the control group. Their median survival was significantly prolonged. It is concluded that double suicide genes are more effective for killing effect on K562 cells in vivo and in ex vivo. It may be applicable to clinical gene therapy.
Cytosine Deaminase ; genetics ; Flucytosine ; pharmacology ; Ganciclovir ; pharmacology ; Genes, Transgenic, Suicide ; genetics ; Genetic Therapy ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Protein-Tyrosine Kinases ; genetics ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; Recombination, Genetic ; Retroviridae ; genetics

Objective To investigate the feasibility of self-made capacitation liquid for in vitro fertilization of Mongolian gerbils, and to provide a reference for gerbil embryo cryopreservation. Methods In vitro fertilization of Mongo?lian gerbil was performed with the self?prepared capacitation solution and semen, and the 2?cell embryos of Mongolian ger?bils were cultured in vitro using an improved KSOM culture medium. Results The in vitro fertilization rate of gerbils was over 60%, and some gerbil 2?cell embryos could develop further in vitro. Conclusions An in vitro fertilization and embry?o development system of Mongolian gerbil has been established, but it needs further optimization.

Objective To explore the technique and clinical efficacy of retroperitoneoscopic nephrectomy (RN) at different anatomical plane for benign non-functioning kidneys. Method We retrospectively reviewed the clinical data of 84 patients who underwent RN for benign non-functioning kidneys from February 2010 to November 2016. Various anatomical plane was performed for differing etiology, image and plane, included radical nephrectomy (A group, n = 35), simple nephrectomy (B group, n = 27) and subcapsular nephrectomy (C group, n = 22). Clinical data and preoperative information was collected and analyzed. Result Nephrectomy was performed successfully in 84 patients. There are significantly longer mean operative time and more estimated blood loss in the C group than that in A group and B group (P < 0.05), however, postoperative hospital stay, complications and recovery time were significantly less in the three groups. There was longer mean operative time and more estimated blood in specific and nonspecific infection group compared with the other two groups (P < 0.05). Conclusion Retroperitoneoscopic nephrectomy, although challenging, is safe, reliable, and successful for treatment of benign non-functioning kidneys.Anatomical plane of avascular zone should be offered as the choice of modality to all patients with benign non-functioning kidney.

OBJECTIVETo report the modified technique and the short-term results of simultaneous pancreas-kidney transplantation (SPK) with the enteric drainage (ED) of exocrine secretions.

METHODSFrom June 2000 to August 2006, thirty-eight patients with diabetes complicated with uremia underwent SPK. The pancreas graft was placed intraperitoneally with exocrine secretions drained into the proximal jejunum without Roux-en-Y procedure. The mean cold ischemic times of pancreas and kidney were (10 +/- 2.0) h and (7 +/- 2.0) h, respectively. Quadruple immunosuppressive therapy with antilymphocyte globulin or anti-CD25 monoclonal antibody, tacrolimus, mycophenolate mofetil and steroids was adopted except one patient.

RESULTSThe 6-month survival rates of patients and grafts were both 97.4% after transplantation. All patients achieved insulin-free euglycemia at (7 +/- 6.9) d postoperative except one. For preoperative patients, mean fasting insulin and C-peptide values were (9 +/- 8.1) mU/L and (6 +/- 4.5) mU/L. After operation, fasting insulin and C-peptide values of patients were (12 +/- 5.8) mU/L and (6 +/- 4.7) mU/L, respectively, which peaked to an insulin level of (57 +/- 43.0) mU/L and a C-peptide level of (11 +/- 6.8) mU/L with stimulation. There were eight cases of delayed renal graft function. All other patients achieved immediate renal graft function. No graft losses occurred due to leakage or intra-abdominal infection. The most common surgical complications were wound infection (n = 12), enteric anastomostic hemorrhage (n = 5) and perirenal hemorrhage (n = 2). Three patients (7.9%) had been reoperated for the reasons of intra-abdominal hemorrhage and perirenal hemorrhage.

CONCLUSIONSSPK is an effective treatment option for selected patients with diabetes mellitus and approaching end-stage renal disease. Enteric exocrine drainage by direct side-to-side anastomosis (without Roux-en-Y) seems to be a simple and reliable technique.

Diabetes Mellitus ; surgery ; Drainage ; methods ; Female ; Follow-Up Studies ; Graft Rejection ; prevention & control ; Graft Survival ; Humans ; Immunosuppressive Agents ; therapeutic use ; Jejunum ; surgery ; Kidney Transplantation ; methods ; Male ; Middle Aged ; Pancreas Transplantation ; methods ; Postoperative Complications ; prevention & control ; Treatment Outcome ; Uremia ; surgery

As a traditional Chinese medicine， Ziziphi Spinosae Semen （ZSS） has the functions of tonifying liver， tranquilizing heart， astringent sweat and producing body fluid， which is used to treat neurasthenia， insomnia， dreaminess， debility， night sweat and so on. With the rapid and constant development of ZSS resource industry and its medicinal value， a large number of by-products and waste generated in the production and processing process， resulting in serious environmental problems. In general， the utilization rate of ZSS resources was still not high. Based on this， the chemical components and potential resources of ZSS were systematically combed from the perspective of the medicinal parts and bioactive components in this paper， and the authors had summarized that the widely application of ZSS and its by-products （fruit， leaf， root， etc.） in the fields of food， medicine， functional food and other areas was discovered and summarized as feed， feed additives， activated carbon， organic fertilizer， etc. In addition， this paper systematically summarized the current environmental protection problems of the industry， and put forward suggestions for improvement， aiming at reducing environmental pollution and improving the utilization efficiency of resources， so as to provide reference and basis for the comprehensive utilization of ZSS and its by-products， and promote the green， economical and double-effect development of the industry.

OBJECTIVETo develop 22 Chinese Mandarin monosyllable lists with good psychometrical equivalence. This study was to evaluate the test-retest reliability of these lists when it was used in speech recognition test in normal hearing dialectal speakers.

METHODSSeven cities including Dalian, Shanghai, Hangzhou, Wuhan, Guangzhou, Fuzhou and Xiamen were selected as testing centers which contain 6 typical Chinese dialectal regions including north of China, East of China, north of Fujian, south of Fujian, Guangdong and mid-south of China. At each center, 22 local normal hearing people were selected to join this study. Every participant was tested by each recognition test of all 22 lists twice in two sessions and same test order respectively. The second run of testing was carried out within 10 days-1 month since first run of testing.

RESULTSThere was a significant correlations between scores obtained at the two sessions (r = 0.682, P < 0.01). Paired student-t test had shown that a gross score of all dialectal participants was significantly higher than that of initial test to retest (P < 0.01). The mean increment of score was (2.7 +/- 10.1)%. A significant difference of test-retest score in 7 sites was 19.8% and it was equal to 5 test items. A one way ANOVA analysis had indicated that there were statistically significant difference between the score improvement of 7 test sites (P < 0.01). Another analysis had shown that there was no significant correlation between test-retest score improvement and intra-session intervals (P = 0.947).

CONCLUSIONSMandarin monosyllabic recognition test seems to be more stable, and the present study has indicated a systematic differences in Chinese Mandarin monosyllable recognition scores between test and retest. Monosyllable recognition test is not susceptible to memory effect. Pearson's correction analysis is not suitable to evaluation for test-retest reliability.

Adolescent ; Adult ; Analysis of Variance ; Asian Continental Ancestry Group ; Audiometry, Speech ; Female ; Humans ; Language ; Reproducibility of Results ; Speech Discrimination Tests ; Young Adult

The present study was designed to characterize the chemical constituents of Guge Fengtong Tablet (GGFTT). Based on the chromatographic retention behavior, fragmentation pathways of chemical components and the published literatures, a diagnostic ion filtering strategy with electrospray ionization quadrupole time-of-flight tandem mass spectrometry (HPLC-ESI-Q-TOF/MS) was established to identify the multiple bioactive constituents of GGFTT. The rapid identification of forty-seven components, including 18 phenolic acids, 8 saponins, 14 gingerol-related compounds, and 7 diarylhepatonoids, was accomplished using this newly developed method. The coupling of HPLC-ESI-Q-TOF/MS with the diagnostic ion filtering strategy was useful and efficient for the in-depth structural elucidation of chemical compounds of GGFTT.
Catechols ; isolation & purification ; Chromatography, High Pressure Liquid ; Diarylheptanoids ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; Fatty Alcohols ; isolation & purification ; Hydroxybenzoates ; isolation & purification ; Saponins ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization ; Tablets, Enteric-Coated ; chemistry ; Tandem Mass Spectrometry

Dao-di herbs, which are widely recognized as medicinal materials with a high quality and good efficacy in clinic, are now facing the dilemma of absence of standard. This study focused on a pivotal scientific problem of design and application of quality standard of Dao-di herbs, and systematically illustrated the general rules for the quality standard of Dao-di herbs involving "four rules, six core contents, and three key methods". The quality standard of Dao-di herbs shall be fully based on literatures as well as habitat, planting/breeding, processing, characters, chemical-pharmacological/toxic data. The common requirements for the quality standard of Dao-di herbs contain "clear source, explicit origin, rational indicator, gradable quality, and multiple detection methods". Notably, traditional experiences and modern techniques, quality tracing management system, "quality determination by distinguishing characters" method, rapid detection technology, effective/toxic substances control method, were comprehensively applied in this standard to purse the objectification, automation, and intellectualization of detection technology. Appearance characters, chemical components, and bioactive parameters, unified effective/toxic indicators, quality markers, and pharmacopeial control indicators and reasonable ranges were included in rigorous quality standards for Dao-di herbs. Besides, simple grading method shall be developed to guide the implementation of "high quality-high price" policy. Eventually, the new quality standards for Dao-di herbs will lead international standards and promote the high-quality development of Dao-di herbs.
Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Plants, Medicinal ; Reference Standards ; Technology

In the long-term evolution, microbes and hosts coexist widely, forming a symbiotic microecosystem and resulting the complex interactions of the metabolism. With the application of microecological theory in Chinese materia medica science, two main points have been accepted gradually. On the one hand, the prevention and treatment of human diseases by traditional Chinese medicines can be achieved through the correction and adjustment of the imbalance of the human microecosystem. On the other hand, the microecosystem can regulate the quality of traditional Chinese medicines in real time, and further affect their curative effect. Thus, a new discipline, Traditional Chinese Medical Microecology, has been gradually established. In this review, the background, theoretical structure, research directions, key problems and the relationship with human microecology of Traditional Chinese Medical Microecology were systematically summarized and prospected for promoting its development. Moreover, this review provides a reference protocol for further discoursing the microecological mechanism involving the efficacy of traditional Chinese medicines.