Practice and reflections on the teaching reform of Experimental Traditional Chinese Medicine guided by scientific research cases / 中华医学教育探索杂志

Experimental Traditional Chinese Medicine is an emerging discipline that plays an important role in cultivating innovative talents of traditional Chinese medicine (TCM). In recent years, with the rapid development of TCM and the new requirements of positioning, and also combined with the college students' cognitive characteristics, we have revised and republished the "Experimental TCM" (Third Edition) textbook, which focuses on introducing knowledge by adopting relevant scientific research cases. This test book was used in the teaching of undergraduates of batch 2013 in the eight-year program in Shanghai University of Traditional Chinese Medicine. After-class questionnaires showed that this teaching mode, guided by scientific research case, is not only helpful for students to develop their quality and ability of adopting modern experimental methods initially in the study and development of TCM, but also able to spread the great achievements of TCM researches. The teaching mode is also conducive to enhancing students' sense of responsibility for the modernization of TCM. Therefore, it is suggested that the course of Experimental TCM should be promoted in the colleges and universities of TCM.

Application and evaluation of moral education materials of scientific research cases in the teaching of Experimental Traditional Chinese Medicine / 中华医学教育探索杂志

Objective:To evaluate the teaching effect of moral education materials implied in scientific research cases in the teaching of "Experimental Traditional Chinese Medicine (TCM)".Methods:The moral education materials implied in scientific research cases of "Experimental TCM" were extracted and skillfully integrated into the teaching of professional knowledge. A questionnaire survey was conducted with questions as "whether it is good to demonstrate the potential humanistic spirit by case teaching, whether this helps improve your interest in science and whether this teaching form affects your study, life and work attitude in the future" "Ten specific items from scientists' moving deeds that touch students and their recognitions" to assess the teaching effect.Results:95.8 percent of students affirmed this teaching form and thought it helped improve their interest in scientific exploration. 87.5 percent of students considered the humanistic spirit would affect their study, life and work in the future. 77.1-89.6 percent of students held a positive attitude to the ten items derived from the scientists' moving stories. Among these items, the percentages of the two items, "the spirit of being able to endure loneliness, work hard to make contributions selflessly without seeking for rewards or reputations" and "having respect forpeople and their scientific research achievements with courage to challenge the authorities of scientific research" are the lowest and highest, respectively.Conclusion:It is good to apply the moral education materials implied in the scientific research cases in the teaching of "Experimental TCM", which basically achieves the teaching goal, but there is still room for improvement.

The Role of Metallothioneins (MTs) in the Occurrence and Development of Hepatocellular Carcinoma and Mechanism Behind the Prevention and Treatment of Traditional Chinese Medicine / 世界科学技术-中医药现代化

It is a brief review of our research on the role of metallothioneins (MTs) in the occurrence and development of hepatocellular carcinoma (Hcc) in more than a decade,and the mechanism of prevention and treatment for Hcc in the field of TCM.It was found that MTs were required for the malignant proliferation of Hcc cells and their expressions significantly increased under different levels of stress.MTs may exert their effects on promoting proliferation via increasing ribosomal protein gene or other gene expressions.The over-expressions of MTs directly altered the balance of ions in the cell nucleus,and interacted on the genes promoting malignant proliferation.Hepatotoxicity and hepatic damage caused by DEN were reduced and relieved to different extents by the representative formula or constitute,which under the common therapeutic principle,by which MTs expressions were down-regulated as well.In conclusion,both effects above are important in the liver protection,inhibition of hyperplasia and carcinogenesis.

Exploration and evaluation of the literature studying and reading route in PBL teaching of the courses of TCM integrated experiments / 中华医学教育探索杂志

Objective On the issues of students' lack of representative collection of literature in problem-based learning (PBL) courses of TCM integrated experiments, to try to explore the litera-ture studying and reading route. Method Totally 41 full-time seven-year medical postgraduate of Shanghai University of TCM were randomly divided into two PBL classes, that are class A (n=20) an class B (n=21). Three PBL units were carried out in class A and class B by parallel; the oriented literatures were provided by tutors alternately, and after PBL courses, the appraisal were performed by students and tutors independently, using SPSS 19.0 for data analysis, t test to compare between groups. Results First, the students' assessment indicated that the 75% (15/20) students of Class A and 80.9%(17/21) students of Class B of class gave a good evaluation of the teaching method. Two classes had more than 80% of the students who thought the literature teachers provided was representative and instructive and over 60.0% students of both classes thought that the teaching measures could improve the classroom efficiency and after-school learning efficiency. The tutors' assessment revealed that there were no significantly differences between the quantity of these references in PPT (P>0.05) made by students in A class and B class in PBL unit 2 and unit 3; but an increasing trend did exist after the leading literatures were provided and the discussions among students were focusing and rele-vant and the teaching goal was achieved . Conclusion The route of providing a small amount of oriented literatures by tutor is positive in PBL courses of TCM integrated experiments, while the design of teaching experiment, the specific way of providing oriented literatures, the quantity and quality of literatures, and even the internal connection between them, all needs further optimization.

Effect of forskolin on corticosterone secretion in Y1 mouse adrenocortical tumor cells / 中国药理学通报

Aim To study the effect of forskolin on cor-ticosterone secretion in mouse adrenocortical tumor cells. Methods Y1 cells were treated with 1μmol· L-1 or 10μmol·L-1 forskolin for 15 min to 24 h. Y1 cells growth morphology was observed, cell culture su-pernatants were collected and corticosterone was tested by ELISA kit. The cells total RNA was extracted using TRIzol kit and was reversely transcribed to obtain the cDNA, then was amplificated by real time quantitative PCR. The cells were lysed with RIPA lysis buffer and protein expressions were carried out by Western blot. Results Y1 cells morphology changed from flat adher-ent to shrink and spherical growth after 1μmol · L-1 forskolin treatment for 3 h. Compared with the control group, corticosterone levels were increased significantly by 1μmol · L-1 forskolin treatment for 24 h ( P <0. 01), then, forskolin significantly enhanced steroid ogenic acute regulatory protein ( Star) mRNA and pro-tein expression ( P <0. 01 ) , moreover, the steroido-genic enzymes such as Cyp11 a1 and Cyp11 b1 mRNA expressions were also up-regulated significantly by fors-kolin ( P <0. 01 ) . Additionally, Star mRNA expres-sion was increased significantly in a time-dependent re-sponse after 10μmol·L-1 forskolin treatment from 1 h to 12 h (P<0. 01). Furthermore, Nr4a1 and Nr4a2 mRNA expressions were up-regulated significantly after 10μmol · L-1 forskolin treatment from 15 min and reached the top at 1 h ( P<0. 01 ) . However, forsko-lin showed no effect on Mc2 r and Nr5 a1 mRNA expres-sions. Conclusion Y1 mouse adrenocortical tumor cells have a strong response to adenylate cyclase ago-nists, then, forskolin can be used to glucocorticoid se-cretion inducer reagent.

Effect of tumor size on the expression of adrenal cholesterol homeostasis molecules in H22 hepatoma-bearing mice / 中国实验动物学报

Objective To study the effect of tumor size on the expression of adrenal cholesterol homeostasis mole -cules in H22 hepatoma-bearing mice.Methods Two hundred and twenty mice were injected with H 22 hepatoma cells to their right armpit.On the 11th day after injection, the mice were sorted according to the tumor size .18 mice with large tumor (large tumor group) and 18 mice with small tumor (small tumor group) were sacrificed, and the tumors were weighed.A control group consisting of 18 normal Kunming mice was also included in this study .The plasma TC, TG and HDL-C were detected using total cholesterol , triglycerides or HDL-C assay kits , respectively .The mRNA expressions of Srb1, Ldlr, Npc1, Npc2, Stard3, Hmgcr, Lipe, Acat1, Abca1, Abcg1, Srebp-1c, Lxrα, Lxrβ, Rxrα, Apoa1 and Apoe were tested by real-time quantitative RT-PCR ( qRT-PCR ) , and Gapdh and β-actin were used for normalization .SRB1 and ApoA1 proteins were analyzed by Western blot .Results The tumor weight was significantly higher in the large tumor group than that in the small tumor group (P<0.05).Compared with the control group , the plasma HDL-C was significant-ly decreased in the two hepatoma groups (P<0.05).The expression levels of Srb1, Ldlr, Apoa1 mRNA and SRB1 protein were significantly increased in the large tumor group (P<0.05 for all).The ApoA1 protein level was significantly higher in the large tumor group than that in the small tumor group (P<0.05).The expressions of Acat1, Lipe, Abca1 and Abcg1 mRNA were significantly lower in the large tumor group than those in the small tumor group (P<0.05 for all).However, the expressions of Srebp-1c, Lxrαand RxrαmRNA were not significantly changed , then, Srebp-1c, Lxrβand RxrαmRNA expressions were significantly up-regulated in the small tumor group (P<0.05).The expressions of Hmgcr and Apoe mR-NA were not significantly different in the two groups .Conclusions In hepatoma-bearing mice , due to the adaptation to tumor-induced chronic stress response , the adrenal cortical cells can effectively utilize intracellular cholesterol to synthetize cortical hormones .

The exploration and innovation ability development of medical students in higher colleges of traditional Chinese medicine under current TCM new situation / 中华医学教育探索杂志

In the new economic situation for the new requirements for traditional Chinese medicine innovation talents cultivation, in view of the relevant medical education at home and abroad experience and many years of exploration in our school, we suggest that in the curriculum construction and development of the national institutions of higher learning of Chinese medicine, the experiment teaching of traditional Chinese medicine should be extended by popularizing college students' exploring experiment and innovative experiment and establishing the combined mechanism between the excel-lent students and excellent teachers. We also put emphasis on developing five abilities, which include the ability of structuring own knowledge system, the ability of acquiring useful knowledge and aban-doning other, the ability of logical reasoning and critical thinking skills, the ability of professional in-terest and innovation, and the ability of expressing themselves and communication skills.

Comparison of the curative effect of three therapeutic regimens on H22 hepatoma-bearing mice / 中国实验动物学报

Objective To compare the therapeutic effect of three treatment regimens on H 22 tumor-bearing mice. Methods H22 tumor-bearing mice were treated by local injection of compound chemotherapeutic agents (5-FU, mitomy-cin and cisplatin ) , oral administration of sorafeni , or the both combined , respectively .Standardized and quantitative syn-drome differentiation methods were used to assess the syndromes and tumor-inhibition rate in the mice .Results All these three therapeutic regimens were effective in suppressing the tumor growth in mice .Among them, the combined therapeutic regimen of local injection of compound chemotherapeutic drugs plus oral administration of sorafeni was the best .Conclu-sion The effect of sorafenib combined with local injection of compound chemotherapeutic agents is better than the other two regimens used separately .

The expression features of hydrolase genes related to the sec retion of thyroid hormone of H22 hepatoma mice with different symptoms in early stage / 国际中医中药杂志

Objective To study the expression features of hydrolase genes related to the secretion of thyroid hormone of H22 hepatoma mice with different symptoms in early stage. Methods Firstly, The quantitative diagnosis and syndrome differentiation methods were used in H22 tumor-bearing mice in early stage, the expression profile of Tg and related hydrolase genes in poisonous pathogenic factors syndrome group (PPFS) and qi-deficiency syndromes (QDS) were got, and the major differential expression were selected. Secondly, the experiment was repeated and ELISA were used to detect T3 and T4 in serum, RT-PCR were applied to detect gene transcription level of genes including Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1. Results ① Based on gene chip, the expression of Tg, Ctsb, Ctsd, Ctsl, Napsa and Tpp1were decreased in the first batch of experiment, the exactly ratio was Tg(0.77 in PPFS;0.84 in QDS), Ctsb(0.83 in PPFS, 0.91 in QDS), Ctsd(0.79 in PPFS;no notable change in QDS), Ctsl(no notable change in PPFS; 0.65 in QDS), Napsa(0.78 in PPFS; no notable change in QDS), and Tpp1 (0.75 in PPFS; no notable change in QDS), respectively. ② T3 and T4 downregulated in PPFS (the T3 value was 1.519±0.162ng/ml, T4 value was 2.194±0.305mg/dl) and in QDS (the T4 value is 4.366±0.727μg/dl) in early stage (P<0.01), especially in PPFS, which was in accordance with the change of Tg in both batches. ③the same trend happened in the validation of Tg(0.22 in PPFS;0.38 in QDS), Ctsb(0.31 in PPFS;0.55 in QDS), Ctsd(0.36 in PPFS;0.78 in QDS) and Napsa(0.24 in PPFS;0.59 in QDS) ,while ctsl(1.24 in PPFS;2.11 in QDS) and Tpp1 (2.85 in PPFS;0.85 in QDS)werethe opposite;even this, the total trend of the expression in QDS was still higher than that in PPFS. Conclusion All the results showed that the thyroid function of H22 hepatoma mice was inhibited in early stage especially in PPFS.

Progress of research in adrenal cortical cells model / 中华内分泌代谢杂志

This paper describes the biological characteristics of human and mouse adrenal cortical carcinoma cell lines and presents research progress made in rat and bovine primary cultured adrenal cortical cells.The effect of angiotensin Ⅱ,potassium,adrenocorticotropic hormone and cAMPon different adrenal cortical cell lines were compared and analyzed,regarding the differences of steroid hormone secreted.The article suggests that it is necessary to develop and establish some ideal adrenal cortex cell models for furtherstudying the mechanisms of steroid hormone synthesis and secretion.

Gene-expression signature of AC-cAMP signal pathway in thyroid of H22 tumor-bearing mice with different symptoms / 国际中医中药杂志

ObjectiveTo observe gene transcription characteristic of AC-cAMP signal pathway in thyroid of H22 tumor-bearing mice with different syndromes.MethodsThe quantitative four diagnosis and syndrome differentiation methods and Affymetrix Gene Chip Mouse Exon 1.0 ST Array were used,thyroid gene expression of normal mice,qi-deficiency syndromes (QDS) and poisonous pathogenic factors syndrome (PPFS) in early stage of H22 tumor-bearing mice were detected.Genes of AC-cAMP signal pathway in gene chips,which related with thyroid hormone synthesis and secretion were selected,and then analyzed their expressive characteristic.Results ①In key genes,TG and Pax8 were down-regulated in early stage,TSHr down-regulated in PPFS while contrary in QDS.②the transcription level of most genes in QDS were slightly higher than in PPFS,Nis,Tpo,AC,Ttf1,Titf2 and Prkaca were included.()Slight changes showed in other genes in this pathway.ConclusionIn AC-cAMP pathway of H22 tumor-bearing mice with different symptoms,some key genes showed similar characteristics including TSHr,AC,Pax8,Ttf1,Tiff2,TG,Nis and Tpo.This suggested that the thyroid is inhibited in mice with PPFS.

Differential gene expressions of steroid- and catecholamine-synthesizing enzymes in adrenal gland of rats with different diseases but the same syndromes / 中西医结合学报

Objective: To study the gene expression characteristics of steroid- and catecholamine-synthesizing enzymes in adrenal gland of spontaneously hypertensive rats (SHRs), Goto-Kakizaki (GK) rats and normal Wistar rats with the same traditional Chinese medicine syndromes. Methods: Sixteen-week-old Wistar rats, SHRs and GK rats were used. By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array, we observed adrenal gland gene expressions in normal Wistar rats, qi deficiency Wistar rats, SHRs with qi deficiency and qi excess, GK rats with qi deficiency and qi excess. Differentially expressed genes of steroid- and catecholamine-synthesizing enzymes and their regulatory factors were analyzed. Results: Thirty-one genes were differentially expressed among all syndromes. Hsd3b6 was down-regulated significantly 6.0-fold in GK rats with qi deficiency syndrome and qi excess syndrome, and Cyp11b2 was up-regulated 1.5 times in GK rats with qi deficiency syndrome. Por, Hsd11b2, and Nr2f6 were up-regulated in all syndromes, and Cyp2c23, Cyp4a3, Cyp4a8 and Cyp2e1 were down-regulated. However, Srd5a1 and Nr4a1 were up-regulated only in GK rats, and Lss was down-regulated only in SHRs. Th was up-regulated 1.5 times in SHRs with qi deficiency syndrome, GK rats with qi deficiency syndrome and GK rats with qi excess syndrome. Ddc was up-regulated 1.5 times in GK rats with qi excess syndrome. Dbh was up-regulated 3.0 times in GK rats with qi deficiency syndrome and qi excess syndrome. However, Comt was down-regulated 1.5 times in GK rats with qi deficiency syndrome and qi excess syndrome, and Mao was up-regulated 1.5 times in SHRs with qi deficiency syndrome and qi excess syndrome. Conclusion: Some genes associated with steroid- and catecholamine-synthesizing pathways were differentially expressed in SHRs and GK rats, and the differentially expressed genes may be related to the development of traditional Chinese medicine syndromes.

Methodology and purposes of establishing mouse and rat models for syndrome differentiation and treatment / 中西医结合学报

Abstract: Methodology of syndrome differentiation and syndrome-based treatment in rats and mice has professional characteristics and caters to the research and development of traditional Chinese medicine (TCM). In this paper, the authors introduced their systematic research in five aspects. 1) Rats and mice can be used to simulate TCM clinical practice. Diagnosis and syndrome differentiation can be done to the rats and mice, and information collected by the four diagnostic methods from the experimental animals meets the requirements of treatment based on syndrome differentiation. 2) Standardized and quantified four diagnostic methods and syndrome differentiation for rats and mice can be established, and are operational and applicable for general use. 3) There exists constitution and syndrome diversity in normal rats and mice. A spontaneous syndrome can develop in diseased rats and mice, and it can be accompanied by or even change to another syndrome, similar to that in human beings. 4) There is a complicated material base for syndromes inferred from the different gene expressions and splices in neuroendocrine-immune network. 5) Individualized treatment based on syndrome differentiation, as well as quantified evaluation and comparison of the treatment efficacy can be done in the rat and mouse models of syndromes. The established methodology and criteria for syndrome differentiation and syndrome-based treatment in rats and mice can be used in the following four research fields: 1) syndrome identification on rat or mouse models; 2) research on the basic theories of TCM, such as the research on the viscera manifestation theory, the material base of syndromes, function mechanisms of the treatment based on syndrome differentiation, and the diagnostics of TCM; 3) study in clinical subject of TCM, such as evaluation and comparison of the efficacy of treatment based on syndrome differentiation, protocol optimization of syndrome differentiation and treatment, and preventive treatment of diseases; 4) study in traditional Chinese drugs, such as the research on properties of Chinese herbal drugs, and pharmacological research on Chinese herbal medicines and formulas.

Differentially expressed genes in adrenal gland of H22 liver cancer mice with different syndromes and in different stages / 中西医结合学报

OBJECTIVE: To reveal the characteristics of gene expression in adrenal gland of H22 tumor mice with typical syndromes and in different liver cancer stages. METHODS: By the quantitative four diagnosis and syndrome differentiation methods and GeneChip Mouse Exon 1.0 ST Array, we observed adrenal gland gene expression in H22 tumor mice with pathogenic factor-toxin predominance syndrome and qi deficiency syndrome in the earlier stage, yang-qi deficiency syndrome in the intermediate stage, and qi-yin-yang deficiency syndrome in the advanced stage. Genes highly expressed and remarkably different were analyzed in this study. RESULTS: A total of seventy-three up-regulated coincident genes and twenty-six down-regulated coincident genes in different stages were investigated in the study. Up-regulated coincident genes included Hp, C3, Anxa1, Procr, C2, Il4ra, Cd14, Ptprc, Cd52, C4b, Eno3, Xdh, Gpx3, and so on. Down-regulated coincident genes included nervous system function-related genes such as Plp1, Mbp, Aldh1a1, Cck, Atn1, genes associated with electrolyte metabolism such as Aldh1a1 and Slc22a17, genes related to signal transduction such as Cxcr4, Spag5 and Stmn3, etc, and genes related to transcriptional control and protein biosynthesis such as Hspa1a, Dnajb1, Thra, Hhex and so on. CONCLUSION: With the development of the tumorigenesis, the symptoms and signs and differentially expressed genes in adrenal gland of H22 tumor mice can be measured. Up-regulated and down-regulated coincident genes may be the features of H22 tumor mice different from those of normal mice.

Effects of Erxian Decoction and its separate prescriptions on the levels of luteinizing hormone and follicle-stimulating hormone in anterior pituitary cells from female rats / 中西医结合学报

OBJECTIVE: To investigate the effects of Erxian Decoction (EXD), a traditional Chinese compound herbal medicine and its separate prescriptions such as Wenshen Yijing Recipe (WSYJR, a recipe for warming kidney and replenishing essence), Ziyin Xiehuo Recipe (ZYXHR, a recipe for nourishing yin and dispersing fire) and Tiaoli Chongren Recipe (TLCRR, a recipe for regulating thoroughfare and conception vessels) and some extracts from EXD on the levels of LH and FSH in the primary cultured anterior pituitary cells from female rats. METHODS: EXD, its separate prescriptions and traditional Chinese herbal extracts from EXD were added directly to the incubators. The levels of LH and FSH were tested by radioimmunoassay. RESULTS: The levels of LH and FSH in the supernatant of anterior pituitary cells treated by EXD or its separate prescriptions including ZYXHR and TLCRR were increased significantly as compared with those in the blank control. There was a tendency for stimulating the secretions of LH and FSH in the WSYJR-treated group too. Dimethyl sulfoxide as the solvent of icariin and curculigoside could interfere with the results. CONCLUSION: EXD and its separate prescriptions such as ZYXHR and TLCRR can increase the levels of LH and FSH significantly.

"Same Disease but Different Syndromes" and "Same Syndrome but Different Diseases" of the Experimental Gastric Ulcer Rat Models Caused by Stress and Other Factors / 浙江中医药大学学报

[Objective] To investigate and discuss if there are special syndromes,same disease but different syndromes and same syndrome but different diseases in the different gastric ulcer rat models.[Methods] Based on reports,we select fourmethods-stress,antifani,apoplon,and alcohol to make gastric ulcer model simultaneously,and set up the normal and control groups.Then we use the standardized and quantified four diagnostics and differentiation methods for mice,which are established to examine and appraise the above models in an objective,dynamic and quantified way.[Result] First,different mice models have different syndromes,such as in the water immersion model,damage to Qi blood Yin and Yang was slight and recovered quickly.The anti-fani model,with typical syndromes of cold in the stomach,is a compound model of Qi,blood,Yin and Yang deficiency;second,there exists syndrome evolution in the disease diversity stage in the same rat;moreover,significant syndrome difference exists for the same disease in different rat.For example,on the second day after the alcohol was given to the rats,there was Yin deficiency,or abundance of Qi,or deficiency in both Yin and Yang to the different individual.Thirdly,as for same syndrome but different disease,similar syndromes appear on the same stage in different diseases,as well as the similar syndrome appear on the different stage in different diseases.For instance,the anti-fani model and the apoplon model have similar syndromes of cold in the stomach,Qi deficiency,Yin and Yang deficiency.[Conclusion] There exist respective syndromes in different gastric ulcer mice models.We can infer that to put into practice"treating the same disease with different methods"and"treating different diseases with same method"will help improve the curative effect.

Effects of Bushendingchuan Decoction on the blood level of CORT and expression of hypothalamic CRHmRNA in asthmatic rats / 中成药

Objective: To study the change of the blood level of CORT and expression of CRHmRNA in asthma-tic rats and the effects of Bushendingchuan Decoction(Cornu Cervi, Herba Cistanches, Radix Morindae Officinalis, Radix Rehmanniae Preparata, etc.) on them. Methods: The change of level in CORT was measured by radioimmunoassay, the change of level in hypothalamic CRHmRNA was measured by RT-PCR(Reverse transcription and polymerase chain reaction by the semi-quantitative determination). Results: The blood level of CORT decreased(P