Exploration on integrated medical education reform in comprehensive universities / 中华医学教育探索杂志

After years of exploration, Central South University has comprehensively reformed and upgraded the curriculum in both horizontal and vertical integration, including optimizing and constructing a basic clinical core curriculum system with organ system integration as the main line, utilizing the advantages of comprehensive universities to further advance the cross-teaching reform of science, engineering, arts and medicine, strengthening pre-medical education with the goal of early exposure to medicine, promoting the reform of early contact clinical integrated teaching according to the concept of "early, multiple and repeated clinical practice", and accelerating the integration of clinical skills training courses with the support of clinical skills simulation teaching. After the integration, the faculty team has gradually matured and the teaching quality has been significantly improved, which has strengthened the students' medical thinking and overall literacy.

An Exploration of the development of medical simulation teaching center / 中华医学教育探索杂志

Medical simulation teaching is an emerging and developing teaching method in our coun-try. At present, the simulation teaching centers are developing rapidly in higher education institutions, but how to improve the effectiveness in the operation of the centers is still a subject that needs to be discussed. Based on our own experience, this paper analyzes the related factors in developing medical simulation teach-ing centers, making summary and demonstrations from aspects of team building, teacher training, docking needs, staffing and so on, so as to provide references and suggestions for the construction of medical simu-lation center higher education institutions.

Effects of silencing HOXA13 gene on malignant phenotypes of hepatocellular carcinoma HepG2and QGY-7703 cells / 吉林大学学报(医学版)

Objective:To investigate the effects of silencing HOXA13 gene on the malignant phenotypes of hepatocellular carcinoma cell lines HepG2and QGY-7703,and to provide a new molecular target for the diagnosis and treatment of hepatocellular carcinoma.Methods:The interference recombinant plasmid of plent-U6-GFP/si-HOXA13 was constructed,then it was stably transfected into the HepG2and QGY-7703 cells.The interfering effects of HOXA13 gene were determined by RT-PCR and Western blotting methods.The HepG2and QGY-7703 cells transfected with HOXA13 knockdown were used as experimental group,and the HepG2and QGY-7703 cells transfected with empty plasmid were used as control group.Then the growth speed was examined by MTT assay, the cell doubling time was examined by double time assay,the colony formation ability was examined by colony formation assay,and the cell cycle was examined by flow cytometry.Results:The MTT assay results showed that compared with control group,the growth speeds of HepG2and GY-7703 cells in experimental group were significantly decreased and the cell cycles were changed;the number of HepG2and QGY-7703 cells in G1phase was increased and the number of cells in S phase was decreased.The doubling time of HepG2and QGY-7703 cells in control group and experimental group were(4.59±0.27),(4.93±0.17),(6.02±0.86),and(6.43±0.66)h, and the differences between control group and experimental group were significant(P<0.05).The colony number of HepG2and QGY-7703 cells in control group and experimental group were 264.00 ± 12.62,269.00 ± 4.55, 165.00± 10.61,and 215.00 ± 4.43,and the differences between control group and experimental group were significant(P<0.01).Conclusion:HOXA13 can increase the proliferation,enhance the clone formation,decrease the number of cells at G1phase and increase the number of cells at S phase in the hepatocellular carcinoma HepG2 and QGY-7703 cells;and it may used as a new molecular target for diagnosis and treatment of hepatocellular carcinoma.

Study on extraction and clathration process of volatile oil in Tianma-Shouwu tablet / 国际中医中药杂志

Objective To optimize the extraction and β-cyclodextrin clathration process of volatile oil in Tianma-Shouwu tablet.Methods The extraction of volatile oil was optimized through the orthogonal design L9(34) with the volatile oil extraction amount as assessment index. The saturated aqueous solution preparation of clathrate form was used. The volume of β-cyclodextrin and volatile oil, clathration temperature, clathration time was investigated by central composite design-response surface methodology, with the score of volatile oil inclusion ratio and utilization ratio as the index. The results were fitted by second-order polynomial equation, and then the best clathration process was determined. Results The optimization extraction process of volatile oil in Tianma-Shouwu tablet was as follows:to add 8 times water,immerse 30 minutes,steam distill 6 hours;and the best clathration process was as follows: the ratio of β-cyclodextrin and volatile oil was 9.50:1, clathration temperature 43 ℃, and clathration time 4.2 hours. Conclusions The optimized extraction and clathration process were reasonable, stabled and repeatable.

Research on HPLC fingerprint ofJinjing solid beverage / 国际中医中药杂志

Objective To establish an HPLC fingerprint ofJinjing solid beverage,and provide experimental evidence of evaluating its quality.Methods Ten batches ofJinjing solid beverage were analyzed by HPLC under the gradient elution condition. A Kromasil C18 column(4.6 mm×250 mm, 5μm) was used, and the flow phase was acetonitrile-H2O(acidified to 0.1% with phosphoric acid) with gradient elution, and the detection wavelength was 327 nm, and the flow rate was 1.0 ml/min, and the column temperature was 35℃. The data were evaluated by the similarity evaluation software for TCM fingerprint.Results The HPLC fingerprint ofJinjing solid beverage were established. Twelve common peaks including chlorogenic acid were identified with similarity of more than 0.9.Conclusion HPLC method is a reliable, available and quick method, that provides a means for controlling and evaluating the quality ofJinjing solid beverage.

Extraction optimization for Yinsangju solid beverage by orthogonal design and Box-Behnken design-response surface methodology / 国际药学研究杂志

Objective To optimize extraction process of Yinsangju solid beverage(YSJ). Methods The extraction of YSJ was preliminarie optimized through the orthogonal design,and then further optimized by Box-Behnken design-response surface with the overall desirability(OD)of chlorogenic acid and polysaccharide content and yield of extract as evaluation indexes. Results The op?timum extraction of orthogonal design was as follow:10 times the amount of water,soaked 0.5 h,extracted 2 times,each time 1.5 h. The optimum extraction of Box-Behnken design was 10.82 times the amount of water,soaked 0.51 h,extracted 2 times,each time 1.77 h. Conclusion The test results show that the Box-Behnken response surface method optimization of extraction process parameters is more precise and more detailed and predictability is good,and the product quality is improved.

Study on the HPLC Fingerprints of Dangua Yangmu Cream / 中国药房

OBJECTIVE:To establish the HPLC fingerprints for Dangua yangmu cream. METHODS:HPLC was performed on the column of Kromasil C18 with mobile phase of acetonitrile-0.026% phosphoric(gradient elution)at flow rate of 0.8 ml/min,the detection wavelength was 270 nm,the column temperature was 25 ℃ and the injection volume was 20 μl. The chromatographic peak of aurantio-obtusin was used as reference peak to determine the 10 batches of Dangua yangmu cream,and Similarity Evalua-tion System for Chromatographic Fingerprint of TCM(version 2.0)was conducted to identify common peaks and evaluate similari-ty. RESULTS:There were totally 25 common peaks in the 10 batches of Dangua yangmu cream,and the similarity was not lower than 0.921. The validation results showed the fingerprints of 10 batches of Dangua yangmu cream had good consistency with the ref-erence fingerprints. COMCLUSIONS:The established method is specific and reliable,and can provide basis for the quality evalua-tion and control of Dangua yangmu cream.

Optimization of Erzhi pill molding process / 国际药学研究杂志

Objective To optimize the molding process of Erzhi pill. Methods Using the external properties of pill, the pill weight, disintegration time as the comprehensive evaluation index, the L9(34) orthogonal test was used to test the effects of three factors:the relative density of thick paste(A), the powder fineness(B), and the proportion of powder and honey(C). Results The best molding process was as follows: the relative density of thick paste was 1.25, the powder fineness was 100 meshes, and the proportion of powder and honey was 1: 0.6. Conclusion This optimized molding process proved to be repeatable, simple and feasible. The Erzhi pill made by this preparation technology are round, the color and luster are uniform and the dissolving time is short. It provides the basis for studying the molding process of Erzhi pill .

Effects of superfine pulverization technology on the dissolution of Erzhi pill / 国际药学研究杂志

Objective To determine the dissolution rate of Erzhi pill which was prepared by using different crushing technologies with specnuezhenide as index , and study the influence of superfine pulverization technology on the dissolution of Erzhi pill. Methods Crude drugs were crushed into the fine powder by employing general crusher and ultra-micro pulverizer,respectively. The water-honeyed pills were prepared by using these fine powders. The method of rotating basket and HPLC were used for the determination of dissolution of specnuezhenide in vitro. The data were analyzed by applying the law of Weibull. Results Compared with Erzhi pill B,the dissolution parameters of specnuezhenide in Erzhi pill A ,including T50、T60、T70、T80、T90、F1h and F2h, showed significant differentce(P<0.05). Conclusion Applying the superfine pulverization technology can improve the dissolution of Erzhi pills in vitro.

Molecular cloning and alternative splicing analysis of hepatoma associated gene HTA / 中南大学学报(医学版)

Objective:To obtain the full length cDNA sequences of hepatoma associated gene HTA, analyze its alternative splicing, detect the expression pattern of 2 HTA gene transcripts in different hepatic cell lines, and to establish a base for further study of HTA gene function in hepatocellular carcinoma (HCC) occurence and development. Methods:The full length cDNA of HTA gene was cloned by rapid amplification of cDNA 3' ends (3'-RACE), rapid ampliifcation of cDNA 5' ends (5'-RACE) and DNA sequencing. The gene structure and alternative splicing were analysed. Northern blot assay was performed to detect the expression pattern of 2 HTA gene transcripts in different hepatic cell lines. Results:The full length of HTA gene was 1414 bp, composed of 3 exons and 2 introns, and the second intron could be retained in mRNA. Northern blot assay showed that 2 transcripts of HTA mRNA(1.4 kb and 1.7 kb) could express in the HCC cell lines HepG2 and QGY-7703, but not in the non-malignant cell line L-02 and HUVEC. The expression level of 1.4 kb transcript was much higher than 1.7 kb one. Conclusion:This study successfully has obtained the full length cDNA of HTA gene, and analysed the gene sequence and alternative splicing, 2 transcripts of HTA mRNA specifically expressed in HCC cell lines. As a hepatoma associated gene, HTA deserves further investigation.

Expression of scFv SA3 against hepatoma fused with enhanced green fluorecsent protein and its targeted ability in vivo / 中南大学学报(医学版)

Objective To express and purify the human scFv antibody,SA3,against the hepatoma fused to enhanced green fluorecsent protein,and to observe the targeted capacity of fusion protein EGFP-SA3 in vivo.Methods SA3 and EGFP genes were cloned into plasmid pET-25b( + )to construct the recombinant plasmid EGFP-SA3/pET-25b ( + ),followed by DNA sequencing.Then it was transformed into E.coli BL21 ( DE3 ) and induced for fusion expression of EGFP-SA3with IPTG.The expressed fusion protein EGFP-SA3 was purified and detected with SDS-PAGE.HepG2 cells were incubated with the fusion protein EGFP-SA3 in vitro,and the binding bioactivity was observed under the fluorecsent microscope.Further more,we injected the EGFP-SA3 by caudal vein into nude mice planted by hepatoma and observed the whole body fluorescence image of EGFP.Results SA3 and EGFP genes were successfully cloned into pET-25b( + ),which was confirmed by restriction enzyme Nco I-Xho I or Nco I-Eco RI.A band migrated at the position 750 bp,same to EGFP gene,emerged when recombinant plasmid was digested by restriction enzyme Nco I-Eco RI.Similarly,a band,about 1 500 bp,emerged when digested by Nco I-Xho I.The open-reading frame was confirmed by DNA sequencing.Fusion protein EGFP-SA3 was expressed as inclusion body.After purification and refolding,the result of immunofluorecsence detection verified that EGFP-SA3could specifically bind to HepG2 cells and maximum tumor penetration was at 24 h after the injection.Conclusion The purified fusion protein EGFP-SA3 has strong binding capacity to HepG2cells,indicating the scFv SA3 has a potential value as a targeting molecule for diagnosis and targeted therapy for liver cancer.

Construction of eukaryotic expression vector of major histocompatibility complex class I -related chain A and establishment of its stable transfected Tca8113-Tb cell line / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To construct the eukaryotic expression vector, encoding major histocompatibility complex class I-related chain A gene (MICA), for the further research of transfecting Tca8113-Tb cell line(a metastatic cell line of brain metastasis from human tongue cancer Tca8113 cells in nude mouse), and to establish a stable MICA overexpression oral squamous cell line.</p><p><b>METHODS</b>cDNA of MICA gene from pCMV-SPORT6-MICA was amplified by PCR, and subcloned into eukaryotic expression vector pEGFP-N1 marked with green fluorescent protein (GFP). The recombinant plasmid was sequenced and transfected into Tca8113-Tb cell line by lipofectamine 2000. After screen culture by G418, stable tranfected Tca8113-Tb cell line was established using definite dilution method. The expressions of GFP protein was viewed directly with fluorescence microscopy and the overexpression of MICA was identified by RT-PCR, real time PCR and immunocytochemistry.</p><p><b>RESULTS</b>The MICA gene was amplified by PCR and then cloned into the vector, whose sequence was identical to that in the GenBank. The transfected cells showed the expression of GFP. And the overexpression of MICA gene in transfected cells was detected by RT-PCR, real time PCR and immunocytochemistry.</p><p><b>CONCLUSION</b>The recombinant eukaryotic expression vector pEGFP-N1-MICA has been constructed successfully and stably expressed in Tca8113-Tb cell line, providing a foundation for further studies on the function of MICA in vitro.</p>

Effect of vacuum sealing drainage and adsorbing material on treatment of large area of skin avulsion wound / 中国组织工程研究

BACKGROUND:Skin flap replantation was poor in patients with large area of skin avulsion due to poor blood transportation and severe infection; therefore,dressing should be changed again and again.OBJECTIVE:To investigate the effect of vacuum sealing drainage technology and absorbing materials on the large area of skin avulsion.METHODS:A total-of-100 patients with large area of skin avulsion were selected from Department of Medical Beauty of Affiliated Hospital of Xingtai Medical College between September 1,2008 and August 31,2009.The injury area ranged from 10% to 50%.All patients were randomly divided into odd day group (n=58,testing group which was treated with vacuum sealing drainage) and even day group (n=42,control group which was treated with dressing treatment).Length of stay,infection rates and healing were compared between the two groups.RESULTS AND CONCLUSION:Length of stay in the testing group was significantly shorter than control group (P<0.05).In the testing group,two cases had baumanii infection and 4 had pseudomonas aeruginosa infection,and the infection rate was 10%.In the control group,7 cases had pseudomonas aeruginosa infection and 5 had MRSA infection,and the infection rate was 29%.The infection rate in the testing group was significantly lower than control group (P < 0.05).In the testing group,46 patients (79%) were cured in the first therapy cycle,and the other 12 cases (21%) were treated with autoallergic free skin transplantation; in the control group,12 cases were cured in the first therapy cycle,and the other 30 patients were treated with autoallergic skin transplantation following granulation tissue growing.The secondary operation time in the testing group was significantly shorter than control group (P<0.05).The results suggested that the effect of vacuum sealing drainage technology and absorption materials application were satisfactory in a large area of skin avulsion.

In vitro anti-tumor immune mechanism of nasopharyngeal carcinoma bispecific anti-idiotype antibody / 中南大学学报(医学版)

Objective To compare the in vitro antitumor immune responses induced by bivalent bispecific anti-idiotype antibody G22-I50 and monovalent anti-idiotype antibody G22 and I50, and explore its possible mechanism. Methods Proteins G22-I50, G22, and I50 were induced and identified by Western blot and ELISA. Peripheral blood monoclear cells (PBMC) were isolated and stimulated with G22-I50, G22, and I50 anti-idiotype antibodies, respectively. MTT assay and LDH release test were employed to examine the proliferation and cytotoxicity of the PBMC. The levels of IFN-γ, IL-2, and IL-4 in the supernatant were detected by ELISA and changes of T lymphocyte subsets were determined by flow cytometry. Results Western blot showed that G22-I50, G22, and I50 had specific binding capabilities to FC2 (Ab1). The activities of G22-I50, G22, and I50 had recovered and these proteins could be used in the in vitro study. The proliferation and cytotoxicity of the PBMC stimulated with G22-I50 were significantly higher than those stimulated with G22 or I50, The level of IFN-γ and IL-2 in the culture supernatant of the PBMC stimulated with G22-I50 was higher than that in the G22 or I50 group, but the level of IL-4 did not increase.Compared with the G22 or I50 group, the proportion of CD4+ and CD8+ T cells and CD4+/CD8+ ratio significantly increased, and the proportion of CD4+CD25+ T cells significantly decreased in the PBMC stimulated with G22-I50. Conclusion G22-I50 has more potent immunogenicity and would enhance specific antitumor effect which might relate to improving PBMC proliferation, inducing the secretion of Th1 type cytokines, activating CD8+T cells, and suppressing the expression of CD4+CD25+ T cells.

A clinical study of 90 patients with pancreatic carcinoma / 中国综合临床

Objective To improve the early diagnosis and prognosis factor of pancreatic carcinoma by summarizing and analyzing the clinical data. Methods The clinical data of 90 patients with pancreatic carcinoma of our hospital from 1989 to 2005 were analyzed retrospectively. Results The symptoms of pancreatic carcinoma were very complicated,the most common manifestations were bellyache,jaundice and weight loss. Main physical signs in these patients included abdominal tenderness,abdominal mass,hepatomegalia,gallbladder enlargement. Jaundice was the outstanding manifestation of pancreatic head cancer. Among all patients,16 cases accepted sugical resection(17.8%),and the 1-year,3-year,5-year survival rate were 22. 2%,11.1% and 2. 2% respectively. Our data showed that the most important prognostic factors which influenced life span were the surgical procedures,tumor size and location,histological differentiation,TNM stage. Conclusions Clinical manifestations of pancreatic carcinoma are related to TNM stage,tumor size and location,histology type,complication disease. Clinical symptoms only provide clue for diagnosis of pancreatic carcinoma. Laboratory and imaging examination will provide objective evidence for further diagnosis and prognosis in pancreatic carcinoma.

Effect of gamma-aminobutyric acid on malignant phenotypes of hepatocellular carcinoma cell line HepG-2 / 中南大学学报(医学版)

OBJECTIVE@#To determine the effect of gamma-aminobutyric acid(GABA) on proliferation and malignant phenotypes of hepatocellular carcinoma cell line HepG-2.@*METHODS@#HepG-2 cells were cultured by routine method, and then treated with different concentrations of GABA. The proliferation of HepG-2 cells was measured through MTT, doubling time and cell cycles by flow cytometry. The malignant phenotypes were investigated by soft agar colony formation assay.@*RESULTS@#Compared with the control group, GABA efficiently stimulated the proliferation of HepG-2 cells in a dose-dependent manner and affected the distribution of cell cycles of HepG-2 cells. The doubling time of the control group and the GABA-treated group were 39.0, 30.6, 30.0, 27.3, 26.6, and 38.2 h, respectively. The colony formation rates were 3.2%, 4.2%, 5.4%, 6.6%,6.5%, and 3.5%, respectively. Tumorigenicity testing showed that the average weights of tumor was 1.382 g, and 0.285 g for the 2 groups. The difference between the control group and the GABA-treated groups was significant (P<0.01).@*CONCLUSION@#GABA can enhance the proliferation and malignant phenotypes of HepG-2 cells.

Screening and characterization of human scFv antibodies against nasopharyngeal carcinoma / 中国医师杂志

Objective To screen the anti-nasopharyngeal carcinoma scFv from a human anti-nasopharyngeal carcinoma single-chain phage antibody library, and identify its characteristics. Methods The single-chain phage antibody library was subjected to three rounds of positive and negative cell panning and enrichment, and then it was selected by ELISA. The binding specificity of phage antibodies with naso-pharyngeal carcinoma cells was confirmed by immunohistochemistry. Results After panning, enrichment and testing by ELISA, 3 phage an-tibody clones reacting with CNE2 more strongly than HUVEC and NP69 were picked out from 4212 clones. One clone, HNSAO33, was fur-ther analyzed after DNA sequencing. The results of immunohistochemistry with cultured cells were similar to those of ELISA. HNSAO33 spe-cifically reacted to nasopharyngeal carcinoma cells in most human nasopharyngeal carcinoma tissue sections except a few human normal naso-pharyngeal tissue sections. The distinction of positive rates was of a great statistical significance. Conclusion ELISA and immunohisto-chemistry results confirmed HNSAO33 specifically bind with nasopharyngeal carcinoma cells. The seFv fragment against nasopharyngeal carci-noma may be further developed and applied in clinical diagnosis and therapy of nasopharyngeal carcinoma.

Construct a subtractive library of Caski cell lines induced by exposing to the space environment by suppression subtractive hybridization / 中国医师杂志

Objectives Construct a subtractive library of Caski cell line induced by exposing to the space environment by suppression subtractive hybridization and pave the way to explain the molecular mechanisms of the changes at the gene level. Methods Super SMART cDNA synthesis and suppression subtractive hybridization (SSH) were performed to isolate differentially expressed cDNA fragments from strains subclonal 48A9 cell line. cDNA from the 48A9 cell line were used as " tester" , and the other from the control Caski cell line as "driver". Subtractive products were directly inserted into T/A cloning vector, and then transformed into host bacteria to set up a subtractive cDNA library of specially or highly expressed genes in strains subclonal 48A9 cell line. Results mRNA were directly extracted and purified with good quality. Double strand cDNA were reverse transcripted integratedly, and then cut by Rsa I into even length short segments. Liga-tion was identified as high effective. After two hybridizations, a subtractive library of differentially expressed genes in strains subclonal 48A9 cell line was successfully constructed by SSH. Conclusion SSH is an effective approach to isolate differentially expressed genes.

Study on effect of ultramicro-shatter technology on penetrating skin absorption of Pollen Typhae in Zhongtongxiao Cataplasm / 中华中医药杂志

Objective:To study the effect of ultramicro-shatter technology on penetrating skin absorption of isorhamnetin-3-O-neohesperidin in Zhongtongxiao Cataplasm.Methods:To apply reformed Frans penetrating skin absorption cell marching extraorgan penetrating skin experiment.HPLC method was used to determine the content of isorhamnetin-3-Oneohesperidin in ultramicro-shatter Zhongtongxiao Cataplasm and in common Zhongtongxiao Cataplasm.Results:The Q-t equation of ultramicro-shatter Zhongtongxiao Cataplasm:Q=3.0382t+47.082,penetrating skin velocity:3.0382(?g.cm2/h);the Q-t equation of common Zhongtongxiao Cataplasm:Q=2.7967t+39.752,penetrating skin velocity:2.7967(?g.cm2/h);Extraction rate of dynamic extracting micro-powder,the ephedrina hydrochloridum,glycyrrhizic acid and glycyrrhizae glycoside were higher than the trdtional cut crude drug decocting.Conclusion:The accumulating osmolality and penetrating skin velocity of isorhamnetin-3-O-neohesperidin in ultramicro-shatter Zhongtongxiao Cataplasm were all better than those in common Zhongtongxiao Cataplasm,it explained that ultramicro-shatter technology accelerate the dissolution of medicine compsitions.

Preparetion of Ketanning dispersible tablet / 中华中医药杂志

Objective: To optimize the formulation of Ketanning dispersible tablet.Methods: The Ketanning dispersible tablet was prepared by using wet granules.The formulation and preparation technology was optimized by using orthogonal design which took the situation of granules,appearance of taablets,the disintegrating time and the tensile strength as indices.Results: The optimized formulation contained,10%MCC,10% PVPP is inner,10% PVPP is outer,1% magnesium stearate.The tensile strength,the disintegrating time were 70N and 3min respectively.Conclusion: It is successful to prepare immediate release tablet.The optimized formulation is rational and stable,the tablet could be released quickly.