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Retroviral Vector Introduced in the Construction of HSV1-tk/GCV Antitumor Suicide-gene Therapeutic System

Biaoyan DU ; Yuhui TAN ; Yingya WU ; Peng ZHAO ; Lian ZHOU ; Yagang ZHAO

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To explore a method for the construction of antitumor suicide-gene therapeutic system. 【Methods】 The thymidine kinase gene of herpes simplex virus type 1 (HSV1-tk) was orientationally cloned into the retroviral vector plasmid (pLXSN) by DNA recombinant technique, and then the recombinant plasmid of pLXSN-tk was identified by restriction endonuclease cutting and DNA sequencing. Introduced by PolyFect Transfection reagent, the recombinant plasmid was transfected into the packaging cell line PT67. Screened by G418, a cell line, which could produce virus stably, was obtained. The virus was transfected into human gastric carcinomatous cell strain SGC-7901 and the transfected SGC-7901 was applied to evaluate an anti-tumor effect. 【Results】 The identification with restriction endonuclease cutting and DNA sequencing showed that HSV1-tk was successfully inserted into the recombined plasmid of pLXSN-tk. The stable cloned PT67/tk was obtained by screening with G418 and then its amount was enlarged by culture, the titer of the PT67/tk solution being 4?10~4 cfu/mL. The anti-G418 cloned cell line SGC-7901/tk was got by infecting SGC-7901 with the virus. Anti-tumor experiment showed that GCV had an obvious toxic effect on SGC-7901/tk but had no effect on SGC-7901, indicating recombinant retroviral HSV1-tk expressed HSV1-tk gene which had biological activity. 【Conclusion】 Cloning HSV1-tk into the retroviral vector is effective in obtaining the recombinant retroviral HSV1-tk which can express HSV1-tk gene, and also effective in establishing an antitumor suicide-gene therapeutic system. This research naturally lays a foundation for further studying of Chinese medicines having synergistic anti-tumor action with suicide gene.

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Characteristics of Naringenin Contents in Exocarpium Citri Grandis

Xujiang YUAN ; Li LIN ; Shengshan ZHU

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To assay the naringenin content in different medicinal materials of Exocarpium Citri Grandis (ECG). 【Methods】 High performance liquid chromatography (HPLC) was applied to determine the naringenin content in different medicinal materials of ECG. The chromatographic conditions were: gradient elution being performed with methanol as mobile phase A and water - acetic acid (61: 4) as mobile phase B, flowing rate being 1.0 mL/min; detecting wavelength at 283 nm. 【Results】 In the rage of 0.088～0.880 ?g, naringenin showed a good linearity with the chromatographic peak area, r=0.9999, average recovery rate was 95.49% and RSD=2.00% (n=5). The naringenin content in 23 different samples of ECG was in the range of 0.1～1.0 mg?g~(-1). 【Conclusion】 It is the first time to establish the HPLC method for the determination of naringenin content in ECG and the method is accurate and reproducible. Naringenin content in varieties of ECG is stable and the difference of naringenin content between Citrus grandis Osbeck. Var. tomentosa Hort. and Citrus grandis (L.) Osbeck is not significant.

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Cloning and Sequencing of Artemisia annua Squalene Synthase Gene and Its cDNA

Liling FENG ; Qingping ZENG

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To clone the squalene synthase gene of Artemisia annua L. for improving the quality and production of Artemisia annua L. by genetic engineering. 【Methods】 PCR amplification, RT-PCR amplification, ligation of the target fragment with a T-vector and sequence analysis of the interested gene were performed. 【Results】 An expected 3590 bp fragment was amplified by PCR and an expected 1257 bp fragment was amplified by RT-PCR. The two cloned fragments were identified by PCR and restriction enzyme digestion respectively. The preliminary sequence data indicated that the results obtained were similar to that from GenBank, and the difference was only found in several base pairs. 【Conclusion】 The squalene synthase gene and cDNA of Artemisia annua L. were successfully cloned and sequenced.

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Effects of Weishu Powder on Acute Liver Injury Induced by Carbon Tetrachloride in Mice

Xukai LIN ; Senyuan XU ; Yipeng ZHANG ; Tingliang ZHENG ; Wei ZHANG

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To investigate the effect of Weishu Powder (WP) on acute liver injury induced by carbon tetrachloride (CCl_4) in mice. 【Methods】 Fifty mice were randomized to normal control group (group A), model group (group B), bifendate pills group (group C), low-dose WP group (group D) and high-dose WP group (group E). The mice in the above groups were fed with the corresponding drugs by gastric infusion for 7 days. On the 8th day of feeding, the mice except those in group A were given 0.5% CCl_4 peanut oil solution (10 mL/kg) by intraperitoneal injection. Sixteen hours later, liver weight, serum level of alanine aminotransferase (ALT), malondialdehyde (MDA) content and necrotic focus amount in the liver were observed. 【Results】 As compared with the normal group, liver weight, serum ALT level and necrotic focus amount were increased in model mice with CCl_4-induced acute liver injury (P

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Effect of Fucongxiang Liquid on Hippocampal Neurons in Rat Models with Vascular Dementia

Shangzhen YU ; Changying ZHAO ; Juan DU

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To investigate the effect of Fucongxiang Liquid (FL) on hippocampal CA1 neurons in rat models with vascular dementia. 【Methods】 Forty SD rots were randomized to four groups: pseudo-operation group, model group, Naofukang group and FL group. Rat models with vascular dementia were established by repeated cerebral ischemia/reperfusion combined with decreasing blood pressure and then were given with normal saline, Naofukang and FL respectively for 15 days. After treatment, the brain of mrs was taken out to make slices for hematoxylin and eosin staining to observe the histomorphological changes of hippocampal CA1 neurons. 【Results】 As compared with the pseudooperation group, a large amount of damaged, degenerative or even necrotic neurons were fotmd in the model group; FL and Naofukang could reduce the damage of neurons to different extents. 【Conclusion】 FL exerts a protective effect on hippocampal CA1 neurons in mice models with vascular dementia.

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Inhibitory Effect of Kaixin Capsule on Gene Expression of Myocardial Immune Inflammatory Factor in Dogs After Resuscitation

Xiaoxing LUO ; Dawei WANG ; Yonghua ZHAO ; Jian LIANG

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To investigate the effect of Kaixin Capsule (KC) on gene expression of myocardial immune inflammatory factor in dogs after resuscitation and to search for an effective way to counteract the progress of systemic inflammatory response syndrome (SIRS) after cardiopulmonary resuscitation. 【Methods】 Fifteen hybrid dogs were randomly divided into three groups, 5 dogs each: pseudo-operation group (group A), model group (group B) and KC group (group C). The dog model of cardiac arrest and resuscitation was established; the dogs were executed and the heart was isolated 180 min after the recovery of autonomic circulation. Myocardial total RNA was extracted and the mRNA expression of objective gene was detected by reverse transcription polymerase chain reaction (RT-PCR). 【Results】 mRNA expression of Toll-like receptor (TLR_4) and tumor necrosis factor ?(TNF-?) in group B differed from that ingroup A (P

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Therapeutic Effect of Focus-Locating Scalp Circum-Needling for Ischemic Stroke

Yanhui LI ; Lping LI ; Guohua LIN

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To observe the therapeutic effect of focus-locating scalp circum-needling for ischemic stroke (IS) at acute stage. 【Methods】 Sixty-one cases of IS were randomized to two groups: group A (n=31) was treated with acupuncture on homolateral scalp region projected by CT-located focus; group B (n = 30) was treated with acupuncture on conventional scalp regions which were in charge of motor and sensory function. The cases were treated with acupuncture once a day for 30 days. The changes of symptoms and signs and the plasma contents of norepinephrine (NE) and epinephrine (E) were observed before and after treatment. 【Results】 Twenty cases were cured, 10 markedly effective, 1 effective and 0 ineffective in group A; 11 cases were cured, 15 markedly effective, 4 effective and 0 ineffective in group B. The therapeutic effect in group A was better than that in group B (P

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Establishment of Diagnostic Criteria of Spleen-Stomach Damp-Heat Syndrome in Chronic Superficial Gastritis

Shaoxian LAO ; Zheng ZHOU ; Wenliang LIN ; Gengxin CHEN ; Zhixin HUANG ; Hong OUYANG

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

【Objective】 To establish preliminary diagnostic criteria of spleen-stomach damp-heat syndrome (SDS) in chronic superficial gastritis (CSG) by disease-syndrome combined differentiation method and modern epidemiological method. 【Methods】 A clinical questionnaire of SDS in CSG was drafted out based on literatures study and experts' survey. Questionnaire investigation was performed in 146 cases of CSG, which consisted of 81 SDS cases and 65 nonSDS cases. Mter the investigation, discriminant analysis was adopted to set up the discriminant functional equations and to perform frequency analysis. According to the results, the diagnostic criteria and the simplified diagnostic criteria of SDS in CSG were established. 【Results】 The preliminary diagnostic criteria of SDS in CSG were as follows: (1) the major symptoms were characterized as yellow and greasy tongue fur, fullness or distension or pain in the stomach, loose stool and poor appetite; (2) the secondary symptoms were characterized as bitter taste and sticky mouth, chest depression, thirst but unwilling to drink, fatigue and nausea; (3) diagnosis could be confirmed by the existence of yellow and greasy tongue fur accompanied by either 2 of the other major symptoms, or by either 1 of the other major and 2 of the secondary symptoms, or by more than 3 of the secondary symptoms. The simplified diagnostic criteria were characterized the co-existence of yellow and greasy tongue fur and fullness or distension or pain in the stomach. 【Conclusion】 The preliminary diagnostic criteria of SDS in CSG are scientific, objective and practicable to some extent.

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Pharmacological Target of Spleen-Strengthening and Qi-Invigorating Herbal Medicine on Small Intestinal Crypt-like Cells

Weiwen CHEN

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

Achievements of pharmacological studies on IEC-6 small intestinal crypt-like cells (IEC-6 cells) were reviewed. Based on the physiological research of IEC-6 cells, the pharmacological experiment model of IEC-6 cells was established and the pharmacological actions of spleen-strengthening and Qi-invigorating herbal medicine (SQHM) on intestinal epithelial stem cells were observed. Thiazolyl blue colorimetry (MTF), reverse transcription polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), high performance liquid chromatography (HPEC) and immune fluorescence labeling technology were applied in the experiments to observe the effect of SQHM on the proliferation, migration and differentiation of IEC-6. Its influences on the expression of villus protein, divalent mental transferrin, interleukin-6 (IL-6) and ornithine decarboxylase (ODC) and on ODC activity and putrescine content were also observed. The results showed that the extracted chemical components of Radix Codonopsis, Rhizoma Atractylodis Macrocephalae, Radix Astragali seu Hedysari and Radix Glycyrrhizae promoted or inhibited the proliferation, migration, differentiation and absorption function of IEC-6 cells by regulating ODC activity and putrescine production. It is indicated that IEC-6 cells are the pharmacological target of spleen-strengthening and Qi-invigorating herbal medicine.

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Experience of Professor Luo Yuankai in Regulating Menstruation, Promoting Gestation and Calming Fetus

Yuzhen ZHANG ; Songping LUO

Journal of Guangzhou University of Traditional Chinese Medicine.2004;0(05):-.

Guided by Professor Luo Yuankai and influenced by his academic thought, the gynecology department of the first hospital affiliated to Guangzhou University of TCM has made great achievements in the research of regulating menstruation, promoting gestation and calming fetus in recent 30 years. It is believed that based on the principle of kidney controlling the reproduction, kidney-tonifying therapy should be the principal method for the treatment of menstrual disease, pregnant disease and infertility which are characterized as deficiency syndrome.

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