SA-gal在肿瘤放免治疗中清除作用的实验研究

研究半乳糖化链霉亲和素 (Streptavidin -gal,SA -gal)和亲和素 (Avidin ,Av)在大肠癌预定位放免显像中对血中标记生物素化单抗的“清除”作用。从荷瘤鼠的尾静脉内注射131I标记的生物素化单抗 ( 131I-bt-McAb)预定位于肿瘤靶位 ,2 4h后 2组实验组分别给予SA -gal和Av ,再过 6h分别行体内分布测定和显像。对照组未给予清除剂。结果表明 ,SA -gal、Av使用后 6h ,血中131I -bt-McAb水平都明显降低 ,肿瘤 血比值明显提高 (对照组、Av清除组、SA -gal清除组的肿瘤 血比值分别为 0 .4 2、2 .0 9、5.2 3,3组之间P <0 .0 5) ;与Av相比 ,SA -gal清除作用更强 ,而且在体内脏器分布上 ,其非特异性摄取也明显地较Av少。实验结果提示 ,SA -gal和Av作为清除剂在肿瘤预定位放免治疗中能有效地加快血中标记单抗的清除 ,使肿瘤靶 非靶比值得以明显地提高 ;两者相比 ,SA -gal优于Av。

Tumor growth inhibition effect of hIL-6 on colon cancer cells transfected with the target gene by retroviral vector

AIM To observe the tumor inhibitory effects bytransfecting IL-6 cDNA into colon cancer cell lineHT-29 with retroviral vector pZIP cDNA.METHODS Human IL-6 gene was reconstructedin retrovirus vector and transfected into incasingcells PA317 by lipofectamine mediated method,the clones of the cells transferred with hlL-6were selected by G418,and targeted HT-29 cellswere infected with the virus granules secretedfrom PA317 and also selected by G418.Test genetranscription and expression level byhybridization,ELISA and MTT assay,etc.Analyze tumor inhibitory effects according to thecell growth curve,plating forming rate andtumorigenicity in nude mice.RESULT Successfully constructed andtransfected recombinant expressing vectorspZIPIL-6 cDNA and got positive transfected celllines.The colon cancer cell line(HT-29 IL-5)transfected with the hlL-5 gene by retroviralvector was established.The log proliferationperiod and the doubling time of this cell line wasbetween 4 to 7 days and 2.5 days according tothe direct cell count,the cell proliferation wasobviously inhibited with MTT assay,the platinginhibitory rate was 50% by plating efficiencytest.When HT-29 IL-6 cells were inoculated intothe nude mice subcutaneously,carcinogenicactivity of the solid tumor was found superior tothe control group and the size of tumor was notsignificantly enlarged.Injection of combinationvirus fluid containing 11.-6 gene intotransplantation tumors could inhibit the growthand development of the tumor.CONCLUSION IL-6 could inhibit the growth andproliferation of colon cancer cells by retroviralvector-mediated transduction.

TREATMENT OF COMPLICATIONS DUE TO PERITONEAL DIALYSIS FOR CHRONIC RENAL FAILURE WITH TRADITIONAL CHINESE MEDICINE

In this paper,the experience in the treatment of complications due to continuousambulatory peritoneal dialysis for chronic renal failure with traditional Chinese medicine(TCM)is reported.Modified Renshen Yangrong Tang(Ginseng Nutrition Decoction)wasused for anorexia and hypoproteinemia;modified Xiangsha Liujunzi Tang(Decoction ofCyperus and Amomum with Six Noble Ingredients)for abdominal pain and distension;modified Da Chaihu Tang(Major Bupleurum Decoction)for peritonitis;modifiedShenling Baizhu San(Powder of Ginseng,Poria and Atractylodes)for diarrhea due toinsufficiency of the spleen with abundance of dampness;Lizhong Tang(Decoction forRegulating the Function of Middle-jiao)and modified Sishen Wan(Pills of FourMiraculous Drugs)for insufficiency of both the spleen and the kidney;Siwu Tang(Decoction of Four Ingredients)added with other drugs for cutaneous pruritus,andGuishao Sijunzi Tang(Decoction of Four Noble Drugs added with Chinese Angelica Rootand white Peony Root)for renal anemia.The therapeutic principles of invigorating theliver and kidney,strengthening the bones and muscles,and promoting blood circulation toeliminate blood stasis were adopted in the treatment of renal osteopathy,and thetherapeutic principles of invigorating the liver and kidney,expelling phlegm and resolvingdampness,and promoting blood circulation to eliminate blood stasis in the treatment ofhyperlipemia.Shen Tekang capsules(capsules for improving the renal function)wasadministered to patients for strengthening the viability and improving the nutrition state,and the recipe for treating renal function failure(both formulated by the authors)forimproving the renal function so as to decrease the frequency and duration of dialysis.

Construction of HBV-specific ribozyme and its recombinant with HDV and their cleavage activity in vitro

AIM To construct the recombinant of HDVcDNA and HBV-specific ribozyme gene byrecombinant PCR in order to use HDV as atransporting vector carrying HBV-specificribozyme into liver cells for inhibiting thereplication of HBV.METHODS We separately cloned the ribozyme（RZ）gene and recombinant DVRZ（comprisingHDV cDNA and HBV-specific ribozyme gene）intothe downstream of T7 promoter of pTAdv-Tvector and studied the in vitro cleavage activityof their transcripts（rRZ,rDVRZ）on target RNA（rBVCF）from in vitro transcription of HBV Cgene fragment（BVCF）.RESULTS Both the simple（rRZ）and therecombinant ribozyme rDVRZ could efficientlycatalyze the cleavage of target RNA（rBVCF）under different temperatures（37℃,42℃ and55℃）and Mg2+concentrations（10 mmol/L,15 mmol/L and 20 mmol/L）and their catalyticactivity tended to increase as the temperaturewas rising.But the activity of rRZ was evidentlyhigher than that of rDVRZ.CONCLUSION The recombinant of HDV cDNAand ribozyme gene had the potential of beingfurther explored and used in gene therapy of HBVinfection.

Effects of Moxibustion at Shenque(CV 8)on Serum IL-12 Level and NK cell Activities in Mice with Transplanted Tumor

The study was to investigate the effect of moxibustion on the serum IL-2, IL-12 levels and NK cell and ascitic tumor cell activities in H-22 mice with tumor and its mechanisms. The method used in the study was that the IL-2 and IL-12 levels were determined by the double antibody method with ELISA, and the NK cell activities by the MTT measure. The results showed that moxibustion elevated the serum IL-2 and IL-12 levels and the NK cell activities of the mice with tumor, and the elevations were of significance as compared with the controls (P<0.05). It is concluded that moxibustion can inhibit the growth of tumor, which is related to the increase of the serum IL-2 and IL-12 levels and the strengthening of NK cell activities.

Kinase domain insert containing receptor promoter controlled suicide gene system selectively kills human umbilical vein endothelial cells

AIM: To study the selective killing of human umbilical vein endothelial cells (HUVECs) by a double suicide gene under the regulation of a kinase domain insert containing receptor (KDR) promoter and mediated by an adenoviral gene vector. METHODS: Human KDR promoter was cloned by polymerase chain reaction (PCR), and two recombinant adenoviral plasmids pAdKDR-CdglyTK, pAdCMV-CDglyTK were constructed according to a two-step transformation protocol. These two newly constructed plasmids were then transfected into 293 packaging cells to grow adenovirus, which were further multiplied and purifi ed. HUVECs and LoVo cells were infected with either of the two resultant recombinant adenoviruses (AdKDR-CDglyTK and AdCMV-CDglyTK) respectively, and the infection rates were estimated by detection of green ? uorescent protein (GFP) expression. Infected cells were cultured in culture media containing different concentrations of 5-? uorocytosine (5-FC) and ganciclovir (GCV), and the killing effects were measured. RESULTS: The two recombinant adenoviral plasmids pAdKDR-CdglyTK, pAdCMV-CDglyTK were successfullyconstructed and transfected into 293 cells. The resultant recombinant adenoviruses infected cells caused similar infection rates; and the infected cells exhibited different sensitivity to the prodrugs: HUVECs infected with AdCMV-CDglyTK and LoVo cells infected with AdCMV-CDglyTK were highly sensitive to the prodrugs, and HUVECs infected with AdKDR-CDglyTK were similarly sensitive but signifi cantly more sensitive than the LoVo cells infected with AdKDR-CdglyTK (P < 0.001). CONCLUSION: Selective killing of HUVECs may be achieved by gene transfer of double suicide gene under the regulation of the KDR promoter. This finding may provide an optional way to target gene therapy of malignant tumors by abrogation of tumor blood vessels. ? 2006 The WJG Press. All rights reserved.

Increased serum and ascitic fluid levels of soluble tumor necrosis factor-p55 in hepatocellular carcinoma patients

Abstract Objective:To explore the levels of serum and ascetic fluid soluble tumor necrosis factor receptor-p55(sTNFR-p55) and understand their clinical implication in primary hepatocellular carcinoma(HCC) pa-tients.Methods:Enzyme-linked immunosorbent assay(ELISA) was used to exmine the levels of sTNFR-p55 in the serum and ascetic fluid in 25 HCC patients and 25 patients with liver cirrhosis(LC).The test was also performed on the serum of 30 healthy subjects who served as control group.To assess the clinical effects of increased serum concentrations of sTNFR-p55 ,four parameters were analyzed by logistic regression.Results:Serum and ascetic fluid levels of sTNFR-p55 in HCC patients were significantly higher than those in LC patients and controls(P=0.001).No significant difference was found between serum sTNFR-p55 levels in the latter 2 groups(P=0.019),and positive correlation between serum levels of sTNFR-p55 and that in ascetic fluid was noted in the 2 patient groups(r=1.000,P<0.001).Levels of the sTNFR-p55 positively correlated with TBIL and AFP in the peripheral blood of HCC patients(r=0.524,P=0.01 and r=0.234,P=0.03,respectively).Conclusion:Increased levels of sTNFRs-p55 in the serum and ascetic fluid could re-flect the abnormal immune status of the HCC patients and may help predict the development of the tumor.

Long-term Effect of TCM Decoctions in Treatment of Nephrotic Syndrome

Fifty-seven cases of nephrotic syndrome were treated with TCM decoctions as accessory treatment for prednisone and cyclophosphamide, and the effects were observed in a follow-up period of 5-15 years. The long-term complete remission rate of 68.4% and recurrence rate of 26.3% in the treatment group were respectively higher and lower than those in the control group (P<0.01, and P<0.01). The results suggested that the TCM decoctions were very helpful in treating this condition.

Gastric cancer induced by N-methyl-N'-nitro-N-nitrosoguanidine in rat with ulcers and expressions of ras and c-erbB2 genes

Objective: To observe the series of pathological changes during the development of gastric adenocarcinoma in ulcerative rats induced by N-methyl-N′-nitro-N-nitrosoguanidine (MNNG), and the expression profile of related oncogenic protein.Methods: MNNG was administered in rats with ulcers due to acetic acid treatment to induce gastric cancer, and the protein expressions of ras and c-erbB2 genes in the ulcer were examined immunohistochemically along with pathological examination.Results: The incidence of gastric adenocarcinoma in the model group reaches 40% (6/15), while none of the rats developed cancer in the control group with ulcers.Positive expressions of the proteins of p21ras and c-erbB2 were observed in the tissues undergoing canceration in the 6 rats of model group, but were not observed in the 5 control rats; p53 protein expression, however, failed to be detected in both groups.Conclusion: A new animal model of gastric cancer has been established in rats with gastric ulcer after MNNG treatment, which may facilitate the pharmacological research of gastric cancer.

Morphological characteristics of the synaptic structures of substance P-immunoreactive terminals in the marginal division of the rat striatum

In the present study,the morphological characteristics of the synaptic structure(MCSS)of the substance P immunoreactive(SPIR)teminals in the marginal division(MrD)of the ratstriatum were studied by using electron microscopy and immunocytochemistry.Four major typesof SPIR symnapses were identified in teh MrD.axodendritic,axo-spinous,axo-axonic,and com-pound.Axo-dendritic and axo-spinous synapses were more common,and a few axo-axonic andcompound synapses wer observed as well.In the postsynaptic target zones of axo-dendritic,andaxo-spinous synapses,there were small or large dendrites or spires.Some synapses with morethan two synaptic components are referred to as compound synapses.Both symmetric andasymmetric SPIR synapses were seen in the MrD.The vesicles in the SPIR presynaptic boutonswere mostly pleomorphic although a few of them were round,The MCSS distinguishes theultrastructure of the MrD from that of the other part of striatum,which suggests that the func-tion of the MrD may be different from that of the rest of the striatum.

Clinical significance of magnetic resonance cholangiopancreatography utilizing half-Fourier acquisition single-shot fast spin-echo in diagnosing bile duct diseases

Objective: To investigate the clinical significance of magnetic resonance cholangiopancreatography(MRCP) utilizing half-Fourier acquisition single-shot fast spin-echo (HASTE) in the diagnosis of bile duct diseases.Methods: Forty-three patients with obstructive jaundice and 4 without were enrolled in this study. The underlying diseases included bile duct calculi ( 13 cases), chronic cholangitis ( 14 cases) malignant tumors ( 18 cases) and congenital biliary cysts (2 cases). All patients underwent examinations with magnetic resonance imaging (MRI) and MRCP,and 39 were also examined with B-type ultrasonography, 33 with CT and 25 with ERCP and PTC. Three-dimensional image reconstruction was performed using volume-rendered technique (VRE) on the basis of the data obtained by MRCP. Results: The biliary calculi were displayed as circular filling defects in MRCP images, with the proximal end of dilated bile duct taking the form of the mouth of a cup. The bile duct of patients with chronic cholangitis showed distal end dilation and thinner proximal end without discontinuity. Interception of the bile ducts was most frequent(72.2%) in cases of malignant bile duct obstruction, in which the ducts may also be mastoid or resembling rat tails.72.2% of the cases had severe dilation of the bile ducts, which occur in only 16.0% of the benign cases, with significant difference between them ( P < O. O1 ). In images of intrahepatic biliary cyst, intmhepatic duct dilated in the shape of a bursa in connection with the duct. By MRCP, 20 malignant obstructions of the bile ducts were identified with 2 misdiagnoses, and in 25 cases of benign obstructions identified by MRCP, only 1 misdiagnoses occurred. Thus MRCP had the sensitivity, specificity and accuracy of 90.0%, 96.3% and 93.6% respectively in discriminating benign andmalignant diseases of the bile ducts, showing a total diagnostic accuracy of 94.0% that was similar to that of ERCP(92.0%) but significantly higher than those of both CT ( 75.0% ) and B-type ultrasonic examination ( 74.0% ).Conclusion: In diagnosing obstructive jaundice, HASTE MRCP is similar to ERCP but better than CT and B-type ultrasonography, with the merits of fast imaging and high resolution as an ideal sequence for MRCP imaging.

Question & Answer

Chinese insect drugs are drastic in nature, capable ofclearing channels and collaterals to promote a freeflow of qi and blood, and effective in someintractable and obstinate diseases due to long-termstagnation of phlegm and blood, which are hard to betreated by ordinary Chinese drugs. In clinic, properuse of insect drugs can often help raise thetherapeutic effects. Some commonly used pairs ofinsect drugs are introduced in the following.

Relationship between estrogen receptor gene polymorphism and clinical indexes associated with coronary heart disease

Objective: To investigate the relationship between estrogen receptor(ER) gene and the clinical indexes associated with coronary heart disease (CHD). Methods: By means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP), we analyzed ER gene polymorphism in 84 CHD patients and 61 healthy subjects and non-CHD inpatients. The clinical indexes associated with CHD were analyzed in relation to the three ER genotypes. Results: There were significant differences in the incidence of hypertension (58.62%), fibrinogen (Fib) concentration (3.5±0.8 g/L), body mass index (BMI, 25.1±3.2), HDL-C concentration (1.0±0.2 mmol/L) between PP genotype group and other genotype groups (P<0.05). Conclusion: ER gene polymorphism may affect ER-mediated cardiovascular protective effect by modulating the expression of ER.

Effects of Yizhi Capsule (益智胶囊) on Learning and Memory Disorder and β-amyloid Peptide Induced Neurotoxicity in Rats

To explore the effects of Yizhi Capsule （益智胶囊, YZC） on learning and memory disorder and β-amyloid peptide induced neurotoxicity in rats. Methods： Various doses of YZC were administered to Sprague-Dawley （SD） rats for 8 consecutive days, twice a day. On the 8th day of the experiment, scopolamine hydrobromide was intraperitoneally injected to every rat and Morris water maze test and shuttle dark avoidance test were carried out respectively to explore the changes of learning and memory capacities in the rats. Resides, after the cerebral cortical neurons of newborn SD rats aged within 3 days were cultured in vitro for 7 days, drug serum containing YZC was added to the cultured neurons before or after β amyloid peptide25-35 （Aβ25-35） intoxication to observe the protective effect of YZC on neurotoxicity by MTT assay and to determine the LDH content in the supernatant. Results： Compared with those untreated with YZC, the rats having received YZC treatment got superiority in shorter time of platform seeking in Morris water maze test, as well as elongated latent period and less times of error in shuttle dark avoidance test. On the cultured neurons, YZC drug serum could effectively increase the survival rate of Aβ25-35 intoxicated neurons and reduce the LDH contents in cultured supernatant. Conclusion： YZC has an action of improving learning and memory disorder, and good protective effect on Aβ25-35 induced neurotoxicity in SD rats. KEY WORDS

Study of a Novel Small Caliber Vascular Graft in a Canine Model with Optical and Scanning Electron Microscopy

A novel biological small-diameter vascular graft was evaluated in a canine model. 3 cm long segments with 4 mm I.D. were implanted end-to-end in the carotid position of 12 dogs for 6 months. Color Doppler sonography was performed at the first week post-operation, and angiography was then administered to 9 grafts at 4th week, 12th week and 24th week respectively to monitor the graft pantency and blood flow characteristics. Vascular samples containing the grafts were collected at 1st week, 8th week, 12th week and 24th week after implantation. Morphological changes of the grafts were observed by optical and scanning electron microscopic (SEM) studies and compared with that of the original prosthesis and the normal host vessel. All grafts were patent throughout the experiment except one graft. Histopathology and SEM demonstrated both a nearly complete inner capsule of varied thickness lining the graft luminal surface and connective tissue adventitia formation at one-week post-operation. The neointima became confluent at 8 weeks and then compact but had no signs of hyperplasia up to 12 weeks; meanwhile on the neointimal surface newly grown endothelial-like cells were migrating from the stoma to the middle portion. The grafts also illustrated endothelialization in many “islands” in the mid-segment luminal surface of the grafts. In addition, the closer distance the cells towards the stoma were, the more morphological similarity the cells with the normal endothelial were. Taken together, the biological vascular graft remained patent for 24 weeks as a carotid prosthesis, characterized by the early and complete neointima formation plus endothelialization starting before 12 weeks post grafting. Therefore, the graft seems suitable for reconstruction of vascular lesions in dogs. Further studies may be carried out to extend the graft application for the clinical use.

Evaluation wiht Computer of the Personnel Needed for Triage and Operation of the Wounded in Nuclear Warfare

When the troops are attacked by nuclear weapons, the number of the wounded and thetype and the condition of the wounds will change with the equivalence and the way of explosion, thenumber of soldiers taking part, the area of the disposition of the troops, and the conditions of protectionof the personnel. Whether the wounded by nuclear weapons who is pouring in enormous amountcan be correctly classified in time has a very important relation in enhancing the effect of the first-aidand the treatment later on. We worked out a programme about the defined types and criteria of thewounded by nuclear weapons beforehand to be stored into the microcomputer. After nuclear cxplo-sion, it is necessary only to input the known data into the microcomputer from the key-board, thecomputer will immediately tell the number of the wounded of various types, the number of peopleand the time needed to perform the triage task and the surgical personnel needed to performthe operations, so that medical supporting programme can be selected or adjusted on time and the ef-ficiency and quality of the triage and first-aid work can be improved.

AN EXPERIMENTAL STUDY BY SINGLE BLIND METHOD ON THE ANTIOBESITY EFFECT OF XIAOPANGMEI

To evaluate the anti-obesity effect of Xiaopang-mei,an experimental study by single method in threegroups of rats was done.The first group were fedwith Xiaopangmei-01,the second group with aplacebo,and the third group were controls.Ascompared with the control and the placebo groups,the rata that had been fed Xiaopangmei for six weeksshowed body weight,food intake,weight of epididy-mis fat pad and the intestinal absorption of glucoseall significantly decreased.The swimming tolerancetest showed they swam longer than the other twogroups.No significant difference was found betweenthe three groups in the growth and developmentindices such as body length,length of femur and tibia,weight of heart,lungs,spleen,kidneys,stomach,smallintestines,gastrocnemius muscle,and adrenal glands.No side effects were found on the liver and kidney.The secretion of insulin was inhibited after the swim-ming tolerance test and the gestrocnemius muscletest was similar for all three groups of rats.Weconcluded that Xiaopangmei is an effective anti-obesity drug.

Frequency domain cardiography and radionuclide ventriculography for evaluation of left ventricular function in coronary artery disease

Fifty-six patients with chest pain underwent selective coronaryangiography,radionuclide ventriculography (RNV) and frequency domaincardiography (FCG) to evaluate left ventricular function in coronary artery dis-ease (CAD).The sensitivity,specificity and accuracy of FCG were 87.5%,77.8%and 82.9%,respectively,which were slightly less than those of RNV (92.5%,88.9% and 90.8%,respectively).The changes of FCG scores were negatively corre-lated with changes of LVEF (r=-0.586,P【0.01),and TS (r=-0.679,P【0.01).These results indicate that FCG may be useful for the evaluation of leftventricular function in patients with CAD.

The effects of scorpion(Buthus martensi Karsch)venom on rat left ventricular function

In order to investigate the cardiovascular effects of the scorpion(Buthus martensiKarsch)venom(BmKv),the left ventricle of the rats was catheterized via the right carotidartery.The LVP,LVEDP,+dp/dt max,Vmax,HR and BP were observed.The results showedthat intravenous injection of the BmKv(60μg/kg),in comparison with the control,elicited obvi-ous hypertension and increase of cardiac contractility,both of which lasted for 1h,while theheart rate had no significant change rand that pretreating the rats with alpha-adrenergic blocker,phentolamine,antagonized the hypertensive effects,but did not antagonize the increase of cardiaccontractility.Pretreatment with beta-adrenergic blocker,propranolol,has no influence on the ef-fects of the venom.It is suggested that the hypertensive effects are due to the activation of al-pha-adrenergic receptor,whereas the increase of cardiac contractility may not be resulted fromthe activation of beta-adrenergic receptor.The BmKv treated with dithiothreitol before injectionhad no cardiovascular effects,indicating that the intact disulfide bridges play a decisive role inthe cardiovascular effects of the BmKv.

Double potentialities of tumoricide and hematopoiesis of human bone marrow cells activated by IL-2 and IL-3

The biomodulative and hematopoietic potentialities of IL-2 and IL-3 activatedbone marrow(ABM)cells from patients with lung adenocarcinoma were studied in vitro.Human bone marrow(BM)cells could be activated by IL-2 in culture for 7d.TheseIL-2 ABM cells had higher cytolytic activities against cells of H 7402 cell line and freshautologous adenocarcinoma cells and maintained the cytotoxicities longer than IL-2 acti-vated peripheral blood lymphocytes(APBLs),a point of possible importance in adoptiveimmunotherapy for cancer patients.The IL-2 ABM cells also had similar number ofBFU-E and CFU-GM to that had fresh BM cells if 1L-3 was added 48h alter IL-2 inculture.The IL-2 and IL-3 ABM cells might be used to eliminate tumor cells and tosupply reconstitutive elements of BM for autologous bone marrow transplantation.