With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals ; Antibodies, Monoclonal ; chemistry ; genetics ; immunology ; Humans ; Hybridomas ; metabolism ; Immunologic Techniques ; methods ; trends

OBJECTIVETo observe the effect of Shen warming Pi strengthening method on expressions of serum T cell subsets (C045+%, C03+%, and C04 +/COB+) in diarrhea-predominant irritable bowel syndrome (IBS-0) rats. Methods An IBS-0 rat model was established referring to AL-Chaer's modeling method combined with tail clamp and intragastric administration of sanna leaf. After modeling 30 SO rats were randomly divided into 6 groups according to random digit table, i.e., the model group, the high, middle, low dose Wenshen Jianpi Recipe (WJR) groups, and the Sishen Pill control group, 6 in each group. A normal control group consisting of 6 SO rats were also set up. Rats in high, middle, low dose WJ R groups were administered by gastrogavage with boil-free WJ R at the daily dose of 3. 100, 1. 550, 0. 775 g/kg, respectively. Rats in the Sis hen Pill control group were administered by gastrogavage with boil-free Sis hen Pill at the daily dose of 0. 736 g/kg. Equal volume of normal saline was given by gastrogavage to rats in the model group and the normal control group. All medication lasted for 2 successive weeks. Rats' general state, expressions of T cell subsets (CD45+%, CD3+%, and CD4+ /CDB+) changes were observed.

RESULTSCompared with the normal control group, expressions of CD45+% and CD3+% increased, but CD4+ /CDB+ decreased with statistical difference (P < 0. 05). Compared with the model group, expressions of CD45+% and CD3+% decreased, but CD4+ ICDB+ increased with statistical difference in high, middle, low dose WJR groups, and the Sis hen Pill control group (P <0. 05). Compared with the Sis hen Pill control group, there was statistical difference in all indices except CD45+ value in the low dose SWPSM group (P <0. 05). Compared with the low dose WJ R group, the expression of CD3+% decreased in high and middle dose WJR groups, and the Sis hen Pill control group; CD4+ /CD8+ increased in the Sishen Pill control group and the high dose SWPSM group (all P < 0. 05).

CONCLUSIONSWJR showed better treatment effect. The mechanism of Shen warming Pi strengthening method might be achieved by regulating expressions of CD45+% and CD3+%, and CD4+ /CD8+ ratios.

Animals ; Drugs, Chinese Herbal ; Female ; Irritable Bowel Syndrome ; therapy ; Leukocyte Common Antigens ; metabolism ; Medicine, Chinese Traditional ; Rats ; T-Lymphocyte Subsets ; metabolism

OBJECTIVETo explore the clinical anesthesia value of transcutaneous acupoint electrical stimulation (TAES) combined with general intravenous anesthesia in endoscopic bilateral thyroidectomy patients.

METHODSTotally 60 patients who underwent endoscopic bilateral thyroidectomy were equally randomly assigned to 2 groups, the treatment group and the control group, 30 in each group. Patients in the treatment group received TAES combined general intravenous anesthesia, while those in the control group received total intravenous anesthesia. Anesthesia was maintained by target controlled infusion of propofolum combined constant speed infusion of remifentanil in the two groups. TAES was maintained from 30 min before anesthesia induction to the end of endoscopic thyroidectomy at bilateral Hegu (L14) and Neiguan (PC6). The mean artery pressure (MAP) and heart rate (HR) were recorded at different time points of anesthesia, i.e., immediately after entry into the operating room (TO), immediately after intubation (T1), 5 min after intubation (T2), 5 min before incision (T3), 5 min after incision (T4), 30 min after inflation (T5), at the end of surgery (T6), 5 min before extubation (T7), immediately after extubation 0 (T8), and 5 min after extubation (T9). The concentration of IL-6 and TNF-alpha were measured at TO, T3, T5, and T6. The target concentration of propofol was also recorded at T3, T4, and T5.

RESULTSThere was no statistical difference in the operation time between the two groups (P >0.05). Compared with TO in the same group, HR at T3-T4 decreased and increased at T8-T9, and MAP increased at T7-T9 in the treatment group; HR decreased at T3 and increased at T7-T9, MAP increased at T1, T5, T7-T9, and MAP decreased at T2-T3 in the control group. IL-6 increased at T5-T6, while TNF-alpha decreased at T6 in the two groups (P <0.01,P <0.05). Compared with the control group at the same time point, HR decreased at T6-T9, MAP decreased at T1, T4, T5, T7-T9, MAP increased at T3, and IL-6 decreased at T5-T6 in the treatment group (P <0. 05). The concentration and the total amount of propofol were significantly lower in the treatment group than in the control group (P <0.01,P <0.05).

CONCLUSIONSTAES could maintain the hemodynamics more stably and inhibit the stress response in endoscopic thyroidectomy. It also reduce the dosage of anesthetics and improve the safety of anesthesia.

Acupuncture Points ; Anesthesia, General ; Anesthesia, Intravenous ; Electric Stimulation ; methods ; Endoscopy ; methods ; Heart Rate ; Hemodynamics ; Humans ; Interleukin-6 ; Piperidines ; Propofol ; Thyroidectomy ; methods ; Transcutaneous Electric Nerve Stimulation ; Tumor Necrosis Factor-alpha

The data mining technique is adopted to analyze characteristics and rules of acupoint and meridian selection of acupuncture-moxibustion for treatment of vertigo at different time periods in the ancient. The data is collected from literature regarding acupuncture-moxibustion from the pre-Qin period to the end of Qing Dynasty, so as to establish a clinical literature database of ancient acupuncture-moxibustion for treatment of vertigo. Data mining method is applied to analyze the commonly used meridians, acupoints and special acupoints in different dynasties, also possible rules are explored. Totally 82 pieces of prescription of acupuncture-moxibustion for treatment of vertigo are included. In the history the leading selection of acupoitns are Fengchi (GB 20), Hegu (LI 4), Shangxing (GV 23) and Jiexi (ST 41) while that of meridians are mainly three yang meridians of foot and the Governor Vessel, especially the acupoints on the Bladder Meridian of foot yangming had the highest utilization rate, accounting for 23.04%. The acupoint selection is characterized by special acupoint, accounting for 80.6%, among which the crossing points are the most common choice. Distal-proximal acupoints combination is the most frequent method. The results indicate that the ancient acupuncture-moxibustion for treatment of vertigo focused on acupoints in the yang meridians, and the specific acupoints play an essential role in prescription; also the principle of syndrome differentiation and selecting acupoints along the meridians could be seen.
Acupuncture Points ; Acupuncture Therapy ; history ; Data Mining ; History, 18th Century ; History, 19th Century ; History, 20th Century ; Humans ; Medicine in Literature ; Moxibustion ; history ; Vertigo ; history ; therapy

OBJECTIVETo evaluate in vivo pharmacokinetics of Ophiopogonis Radix polysaccharide MDG-1 oily suspension injection prepared with different prescriptions in rats, and explore the feasibility of the long-acting drug delivery of MDG-1 Injection by using the oily suspension drug release system.

METHODMDG-1 microparticles were prepared by the anti-solvent precipitation method. Their size and size distribution were characterized. Castor oil with a high viscosity or aluminum stearate were added into soybean oil with a low viscosity, in order to prepare oily media with different viscosities, detect their rheological properties and screen out superior prescriptions for in vivo evaluation.

RESULTThe average size of microparticles was 21.81 microm, and the span between them was 2.63. The in vivo evaluation was conducted for prescriptions of mixed oil (soybean oil/castor oil, 2: 3) and soybean oils gelled by 2% and 4% aluminium stearate. Among them, the prescription of soybean gelled by 4% aluminium stearate could significantly reduce C(max) and prolong the apparent t1/2, with the MDG-1 release time of several days.

CONCLUSIONIt is feasible to achieve the long-acting MDG-1 drug delivery by using oily media with a high viscosity.

Animals ; Chemistry, Pharmaceutical ; methods ; Drug Delivery Systems ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; pharmacokinetics ; Male ; Ophiopogon ; chemistry ; Plant Oils ; chemistry ; Polysaccharides ; administration & dosage ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Viscosity

Objective:To compare the modified Qjng Long Bai Wei needling method and ordinary acupuncture method in the effects of improving the levels of interleukin (IL)-1β,IL-6 and tumor necrosis factor (TNF)-α in the treatment of knee osteoarthritis (KOA),and to determine the advantage of the modified Qing Long Bai Wei needling method for KOA.Methods:One hundred KOA patients were randomized into a treatment group and a control group by using the random number table,with 50 cases in each group.The treatment group was intervened by the modified Qing Long Bai Wei needling method,and the control group was given ordinary acupuncture.The two groups were observed before and after the treatment to determine the changes in the levels of IL-1β,IL-6 and TNF-α in synovial fluid,and the clinical efficacies were compared between the two groups.Results:The total effective rate and clinical recovery rate were 97.9％ and 52.1％ respectively in the treatment group,versus 85.1％ and 25.5％ in the control group,and the between-group differences were statistically significant (both P＜0.01).After the treatment,the levels of Ib1β,IL-6 and TNF-cα in synovial fluid changed significantly in both groups (all P＜0.01);there were significant differences in the levels of IL-1β,IL-6 and TNF-α in synovial fluid between the two groups (all P＜0.01).Conclusion:The modified Qjng Long Bai Wei needling is an effective method for KOA and it can significantly improve the levels of IL-1β,IL-6 and TNF-α in synovial fluid.

OBJECTIVEIBS-D rat model was established to assess the effect of Shen warming Pi strengthening method (SWPSM) for intervening diarrhea-predominant irritable bowel syndrome (IBS-D) by observing rats' general state, stool properties, AWR ranking, and histopathological changes.

METHODSTotally 72 rats were randomly divided into 6 groups, i.e. the normal group, the model group, the high, middle, low dose SWPSM groups, and the control group, 12 in each group. The IBS-D rat model was successfully established referring to AL-Chaer ED's modeling method. After modeling high, middle, and low dose SWPS Recipe boil-free granules were given by gastrogavage to rats in corresponding treatment groups. Sishen Pill boil-free granule was given by gastrogavage to those in the control group. Equal volume of normal saline was given by gastrogavage to rats in the model group. The medication lasted for 2 weeks. Rats' general state, stool properties, abdominal withdrawal reflex (AWR) ranking, and histopathological changes were observed.

RESULTSAfter treatment, the general state of all rats got im- provement to various degrees. The improvement in the high and middle dose SWPS Recipe groups were superior to that in the low dose SWPS Recipe group and the control group (P < 0.05). There was no statistical difference in the growth rate between after and before treatment in each group (P > 0.05). Compared with the model group and the low dose SWPS Recipe group, the defecation amount within 4 h was less in the high and middle dose SWPS Recipe groups and the control group (P < 0.05). The Bristol ranking score, average ranking of loose stool, ratio of dry stool and wet stool were lower in the high and middle dose SWPS Recipe groups than in the control group and the low dose SWPS Recipe group (P < 0.05). The AWR ranking score was lower in the high and middle dose SWPS Recipe groups than in the control group when the volume of balloon dilation was 1.5 mL. There was no organic change of histological or morphological observation.

CONCLUSIONSHigh sensitive IBS-D model was proved to be reliable. SWPSM could reduce the quantity of stools, lower Bristol ranking score, average ranking of loose stools as well as ratios of dry stool and wet stool, contributing to reducing the high sensitivity of rats' visceral organs to some extent.

Animals ; Diarrhea ; drug therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Irritable Bowel Syndrome ; drug therapy ; Male ; Phytotherapy ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the nephrotoxic effects of methyl cantharidimide tablets on urinary protein and enzymes in Beagle dogs.

METHODBeagle dogs were randomly divided into negative control group(blank tablet), methyl cantharidimide tablets group (6.11,12.21, 24.42 mg x kg(-1)), continuously 30 days of oral adminiStration, once a day. The drug and control group were collected and determined fresh urine in 1, 2, 3 and 4 weeks of the administration; Serum urea nitrogen (BUN), creatinine (Crea), total protein (TP) and albumin (ALB) as well as sodium, potassium, chloride electrolyte were determined on 15 and 30 days of the administration; Urine albumin (mAlb), kidney injury molecule-1 (KIM-1), neutrophil gelatinase-associated lipocalin( NGAL), N-acetyl-beta-D-glucosaminidase (NAG), clusterin, beta2-microglobulin (beta2-MG), alpha1-microglobulin (alpha1-MG), alanine aminopeptidase( AAP) and im- munoglobulins IgG were tested on 15 and 30 days of the administration.

RESULTCompared with the control group, urine protein and white blood cells was significantly increased in each dose group. On 15 days of the administration, mAlb were higher in each dose group, KIM-1, NGAL, clusterin, NAG and AAP were significantly higher in high-dose group, while the middle and low dose group had no significant difference, as well as blood SCr and BUN no obvious abnormalities. On 30 days, mAlb, KIM-1, clusterin, NAG, AAP were increased in each dose group, appearing dose-effect relationship, beta2-MG and NGAL levels were significantly increased in high-dose group. Contents above indicators were increased with significant dose and time relationship, and serum BUN, Scr were correlated, suggesting that urine mAlb, KIM-1, clusterin, NAG and AAP indicators that can sensitively respond the changes of proteins and enzymes in urine.

CONCLUSIONMethyl cantharidimide tablets has a renal toxicity, urine mAlb, KIM-1, clusterin, NAG and AAP can be used as the early nephrotoxic biomarkers of methyl cantharidimide tablets.

Animals ; Biomarkers ; urine ; Dogs ; Female ; Kidney ; drug effects ; Kidney Diseases ; chemically induced ; Male ; Proteins ; metabolism ; Tablets ; adverse effects ; Urine ; chemistry

OBJECTIVETo investigate the role of nitric oxide in the inhibitory effect of somatic afferent input on the pressor response caused by electrical stimulation of the paraventricular nucleus of the hypothalamus (PVN).

METHODSSD rats anesthetized by urethane were used in the study. Bipolar stainless stimulating electrode was inserted into PVN for electrical stimulation. Multi-barreled micropipettes were used for microinjection of L-NAME or normal saline into the lateral ventricle or amygdala. Deep peroneal nerve (DPN) was stimulated with electrical current pulses of 0.4 mA with duration of 0.5 ms at 4 Hz for 5 min. PVN was stimulated by electrical current pulses of 0.3 mA with duration of 0.5 ms at 80 Hz for 10 sec.

RESULTElectrical stimulation of PVN increased mean arterial pressure. Stimulation of DPN significantly inhibited the pressor response induced by stimulation of PVN (P<0.01), with the inhibitory percentage of 43.27%. Microinjection of L-NAME (0.5 mol/L,10 microl) into the lateral ventricle of brain attenuated the inhibitory effect of DPN. The inhibitory percentage decreased from 47.73% to 12.49% (P<0.05). Microinjection of L-NAME (2 mol/L,100 nl) into amygdala reduced the inhibitory effect of DPN. The inhibitory percentage of stimulating DPN on the pressor response decreased from 50.71% to 25.30% (P<0.05).

CONCLUSIONNitric oxide in the brain and amygdala are involved in the inhibitory effect of somatic afferent input on central pressor response.

Afferent Pathways ; drug effects ; physiology ; Amygdala ; drug effects ; physiology ; Animals ; Blood Pressure ; physiology ; Electric Stimulation ; Enzyme Inhibitors ; pharmacology ; NG-Nitroarginine Methyl Ester ; pharmacology ; Nitric Oxide ; antagonists & inhibitors ; physiology ; Nitric Oxide Synthase ; antagonists & inhibitors ; Paraventricular Hypothalamic Nucleus ; drug effects ; physiology ; Peroneal Nerve ; physiology ; Pressoreceptors ; drug effects ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the relationship between genotype of hepatitis B virus and hepatitis B virus related-glomerular nephritis in (HBV-GN) children.

METHODTotally 176 HBV-DNA positive children with chronic hepatitis B were randomly collected. Among the 176 patients, 92 were HBV carriers, 84 were cases with chronic hepatitis. The genotypes of their serum HBV, liver function, and HBV-DNA load were detected. When children showed nephrotic syndrome, renal biopsy was performed.

RESULTOf the serum samples of 176 cases, 85 (48.3%) were genotype C, 72 (40.9%) were genotype B, 13 (7.4%) were genotype B/C, and 6 (3.4%) were non-B/C genotype which were excluded. Among the analyzed 157 cases, the ratio of HBV-GN in the HBeAg positive group (78.3%) was significantly higher than that in the negative group (21.7%) (χ(2) = 18.301, P < 0.001). And, the ratio of HBV-GN in the genotype C group (73.9%) was significantly higher than that in the genotype B group (26.1%) (P < 0.039). The ratio of hematuria or proteinuria in the genotype C group (20%, 18.8%) was significantly higher than that in the genotype B group (8.3%, 5.6%) (P < 0.039; P value = 0.013); and the alteration of ALT or C3 in the genotype C group (10.2%, 15.3%) was more frequent than those in the genotype B group (2.8%, 2.8%) (P = 0.005; P = 0.008). There were no significant differences in kidney dysfunction or hepatomegaly. Further, the ratio of HBV-GN was more significantly frequent in HBV-DNA highly loading group (79.2%) than which in HBV-DNA lowly loading group (20.8%) (P = 0.000). Finally, in HBV-GN group, genotype C cases (88.2%) more frequently had high HBV-DNA load condition than genotype B cases (11.8%) (P = 0.021).

CONCLUSIONChildren with HBV infection in Gansu province showed mainly genotypes C or B, while genotype C seemingly predominant. Patients with genotype C more frequently showed proteinuria or hematuria. The high HBV-DNA load may be related with HBV-GN. It is a potential reason in the mechanism of HBV-GN that patients with genotype C had more possibility to have HBV-DNA high load. Analysis of HBV genotype for HBV patients maybe helpful in diagnosis and treatment.

Adolescent ; Biopsy, Needle ; Child ; Child, Preschool ; China ; epidemiology ; DNA, Viral ; blood ; genetics ; Female ; Genotype ; Hepatitis B ; blood ; epidemiology ; virology ; Hepatitis B virus ; genetics ; Humans ; Infant ; Male ; Nephritis ; epidemiology ; pathology ; virology ; Viral Load