The method of monoclonal antibody-based immunoassay has a great importance in the study of quality control of traditional Chinese medicine (TCM) and detection of trace components in vivo animals. Synthesis of small molecule artificial antigen is the prerequisite for the establishment of this method. In present study, catalpol-BSA was synthesized by sodium periodate oxidation method. Matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF-MS) and molecular exclusion chromatography showed that catalpol was successfully conjugated with BSA. The mice could specifically produce anti-catalpol antibodies with titer up to 1:8000. The artificial antigen of catalpol was successfully synthesized.
Animals ; Antibodies ; immunology ; Antigens ; chemistry ; immunology ; Immunoassay ; Iridoid Glucosides ; chemistry ; immunology ; Male ; Medicine, Chinese Traditional ; Mice ; Mice, Inbred BALB C ; Serum Albumin, Bovine ; chemistry ; immunology

Objective: To observe the effects of acupoints, cone numbers and durations of moxibustion with different moxibustion methods on skin surface and inside temperature, and to provide references for the clinical standardization of moxibustion amount. Methods: The 42 big-ear white rabbits were divided into 6 groups according to the random number table method, a 1-cone direct moxibustion group, a 2-cone direct moxibustion group, a 3-cone direct moxibustion group, a 1-cone herbal cake-partitioned moxibustion group, a 2-cone herbal cake-partitioned moxibustion group, and a 3-cone herbal cake-partitioned moxibustion group, with 7 rabbits in each group. Shenque (CV 8), Shenshu (BL 23) and Zusanli (ST 36) were used in each group, but the moxibustion methods, cone numbers and durations of moxibustion were different. Rabbits in each group received moxibustion once every other day for 5 times in total. During the intervention, a thermoelectricity coupled probe and a temperature recorder were used to record the real-time acupoint skin temperature and the temperature at different time points, so as to observe, analyze and process the real-time changes in the temperature difference between the surface and inside of acupoint skin. Results: For herbal cake-partitioned moxibustion, the best temperature for cone changing was (46.38±0.51) ℃ when the highest surface temperature was (49.20±0.52) ℃; the multi-factor comparison of acupoint × cone number × time and acupoint × moxibustion method × time showed that time × acupoint, time × moxibustion method and cone number × acupoint had interactive effects (all P<0.05). Comparing skin temperature differences between different cone numbers at the same acupoint, Shenque (CV 8) on the 1st and the 5th days, Shenshu (BL 23) on the 3rd and the 7th days, Zusanli (ST 36) on the 1st and the 9th days of experiment showed statistically significant differences (all P<0.05). The skin temperature comparison of different moxibustion methods at the same acupoint all had statistical differences (all P<0.05), except for Shenque (CV 8) before moxibustion, Shenshu (BL 23) before moxibustion and on the 5th day; Zusanli (ST 36) only showed statistical differences on the 5th and 7th days (both P<0.05). The skin temperature differences of different acupoints after moxibustion in the 1-cone, 2-cone and 3-cone groups were statistically different (all P<0.05); direct moxibustion and herbal cake-partitioned moxibustion at different acupoints were all statistically different (all P<0.05). Conclusion: Cone changing temperature under the same specifications of herbal cake-partitioned moxibustion was confirmed. Temperature difference between surface and inside of different acupoint skin at the same maximum temperature was significantly different due to the cone numbers and moxibustion methods, which showed the highest at Shenshu (BL 23), the second at Shenque (CV 8), and the lowest at Zusanli (ST 36). The influence of acupoint factor should be considered to determine the quantitative indicators of moxibustion.

Objective To observe the clinical efficacy of withdrawal therapy based on regulating nutritive qi and defensive qi(shortened to Tiaohe Yingwei method)in treating sedative-hypnotic dependent insomnia of disharmony between nutritive qi and defensive qi type.Methods Ninety patients with sedative-hypnotic dependent insomnia of disharmony between nutritive qi and defensive qi type were randomly divided into the treatment group and the control group,with 45 patients in each group.The control group was given oral use of Estazolam by 25%of weekly dose-reduction,while the treatment group was treated with Chinese medicinal decoction of Tiaohe Yingwei Zhumian Prescription based on Tiaohe Yingwei method together with Estazolam.The treatment course for the two groups lasted for 4 weeks.The changes of Pittsburgh Sleep Quality Index(PSQI)scores,total TCM syndrome scores,and Drug-withdrawal Syndrome Scale(DWSS)scores in the two groups were observed before and after treatment.After treatment,the efficacy for improving sleep efficiency value(IUSEV)and clinical safety in the two groups were evaluated.Results(1)During the trial,2 cases fell off in the treatment group,and 43 cases included in the statistics;3 cases fell off in the control group,and 42 cases included in the statistics.(2)After 4 weeks of treatment,the total effective rate for improving IUSEV of the treatment group was 88.37%(38/43),and that of the control group was 61.90%(26/42).The intergroup comparison by non-parametric rank-sum test showed that the efficacy for improving IUSEV in the treatment group was significantly superior to that in the control group(P＜0.05).(3)After treatment,obvious reduction was shown in the overall PSQI scores and the scores of the items of sleep quality,time for falling asleep,sleep time,sleep efficiency,sleep disorder and daytime dysfunction in the two groups when compared with those before treatment(P＜0.05).The intergroup comparison showed that except for the items of sleep disorder and daytime dysfunction,the treatment group had stronger effect on decreasing the scores of the remaining items and the overall PSQI scores than the control group(P＜0.05).(4)After treatment,the total scores of TCM syndromes of both groups were significantly decreased compared with those before treatment(P＜0.05),and the decrease of the total scores of TCM syndrome in the treatment group was significantly superior to that in the control group(P＜0.05).(5)After treatment,the total DWSS scores of the two groups were significantly decreased compared with those before treatment(P＜0.05),and the effect on lowering the scores in the treatment group was significantly superior to that in the control group(P＜0.05).(6)During the course of treatment,no significant adverse reactions occurred in the two groups,or no abnormal changes were found in the safety indexes such as routine test of blood,urine and stool,liver and kidney function,and electrocardiogram of the patients.Conclusion Withdrawal therapy based on Tiaohe Yingwei method exerts certain effect for the treatment of sedative-hypnotic dependent insomnia of disharmony between nutritive qi and defensive qi type.The therapy is effective on improving the quality of sleep and reducing the incidence of drug-withdrawal syndrome,and has a high safety.

BACKGROUNDIt is well recognized that meteorological factors have important infuences on the onset and development of many kinds of diseases. The present study was undertaken to investigate the effects of the meteorological elements on admission rates of cerebral infarction patients with hypertensive nephropathy at Changchun city, Jilin Province, northeast China.

METHODSA total of 763 medical records of inpatients from nine hospitals at Changchun city, during a period from April 6 to April 17 in 2010, were reviewed. These patients were admitted to hospitals due to the occurrence of cerebral infarction. The hypertensive nephropathy was evidenced with certain diagnosis of essential hypertension and hypertension-related kidney injuries. The cerebral infarction was diagnosed according to the World Health Organization (Stroke) standard. All the meteorological data were from practical monitoring records in Jilin Province Meteorological Observatory. The relationships between the epidemiological prevalence of cerebral infarction and meteorological variables were analyzed using the time series models of statistics.

RESULTSCompared with admission rates before the violent change in meteorological status (April 6 to April 17, 2010), the number of admission patients suffering from cerebral infarction remarkably peaked on April 12. Such an increase was highly correlated with heavy precipitation, elevation of daily average relative humidity, and reduction of average daily air temperature. With the betterment of the meteorological conditions on April 17, the admission rates of cerebral infarction patients dropped to the same level as the dates before snowing (April 6 to April 11).

CONCLUSIONSThe meteorological changes are highly associated with the occurrence of cerebral infarction in patients with hypertensive renal injury in northeast China. This study also suggested that an intensive medical interference for those patients with hypertension-induced organ injuries is very necessary in preventing the occurrence of cerebral infarction with hypertensive nephropathy when there is a violent change in meteorological condition.

Adult ; Aged ; Aged, 80 and over ; Cerebral Infarction ; China ; Female ; Hospitalization ; statistics & numerical data ; Humans ; Hypertension, Renal ; Male ; Meteorological Concepts ; Middle Aged ; Nephritis ; Patient Admission ; statistics & numerical data

OBJECTIVETo identify polymorphisms of the serotonin transporter(5-HTT) gene and to find out whether there was relationship between any such polymorphisms and sleep apnea syndrome (SAS).

METHODSFor two polymorphisms of 5-HTT target DNA gene was amplified using polymerase chain reaction (PCR) and 6% non-denaturing polyacrylamide gels electrophoresis. The frequencies of the different forms of the genotypes and alleles of 5-HTT gene were analyzed in 104 patients with SAS and 150 healthy controls.

RESULTSThe frequencies of the S or L alleles and the S/S, S/L or L/L genotypes in promoter region of 5-HTT gene in SAS group were not significantly different to those in healthy controls (P > 0.05). However, the frequencies of 10/10, 12/10 genotypes of 5-HTT-VNTR in SAS patients were significantly higher than those in healthy control subjects (P < 0.05). Moreover, the frequency of the allele 10 of 5-HTT-VNTR in SAS patients was significantly higher than that in healthy controls (P<0.01).

CONCLUSIONThe allele 10 of 5-HTT-VNTR might be a susceptible factor in the pathogenesis of SAS.

Adult ; Aged ; Alleles ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Male ; Middle Aged ; Minisatellite Repeats ; genetics ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Promoter Regions, Genetic ; genetics ; Serotonin Plasma Membrane Transport Proteins ; genetics ; Sleep Apnea Syndromes ; genetics ; Young Adult

No abstract available.
Diagnosis* ; Fistula*

The purpose of this study was to explore the effect of proteasome inhibitor, bortezomib (Bzb), on osteoblast in pathologic status of myeloma bone disease. The myeloma bone disease was modeled by co-culture of mouse myeloma cell RPMI8226 with osteoblast line MC-3T3E1 from mouse calvaria, and intervenient culture of supernatant. The inhibitory effect of Bzb on proliferation of MC-3T3E1 assayed by modified MTT method, the apoptosis of MC-3T3E1 cells was determined by flow cytometry with Annexin V/PI staining, the expressions of osteoblast markers, Runx2/cbfa1, osteocalcin (OCN) and osterix (OSX) in MC-3T3E1 treated with Bzb were detected by RT-PCR and Western blot respectively. Experiments were divided into 3 group: single cultured, co-cultured and supernatant-interveniently cultured groups. The results showed the Bzb in higher concentration inhibited proliferation of MC-3T3E1 cells in a dose-dependent manner, with the IC(50) of 38.1 nmol/L for 48 hours, the Bzb in low concentration (5 nmol/L) did not show the inhibitory effect on proliferation of MC-3T3E1 in single cultured group (p>0.10), but could decrease apoptotic rate of MC-3T3E1 by 32.5% and 24.6% respectively in cocultured and supernatant-interveniently cultured groups, moreover increased the expression of osteoblast-related gene OSX, OCN mRNA and protein (p<0.05), while no obvious change of Runx2/cbfa1 expression was observed (p>0.05). It is concluded that the proteasome inhibitor, Bzb, in low concentration promotes the activity of osteoblast internal mechanisms, and prevents the apoptosis of osteoblasts induced by myeloma cells. In addition, it can up-regulate transcription and expression of osteoblast markers related to Runx2/cbfa1 path way, thus may protect osteoblasts in myeloma bone disease.
3T3 Cells ; Animals ; Apoptosis ; drug effects ; Boronic Acids ; pharmacology ; Bortezomib ; Cell Line, Tumor ; Mice ; Multiple Myeloma ; pathology ; Pyrazines ; pharmacology

OBJECTIVETo investigate the effect of wortmannin on endothelial cells proliferation and migration induced by high glucose Müller cell conditioned medium (HGMCM).

METHODSImmunofluorescence, flow cytometry and Boyden chamber migration models were used to analyze the effect of wortmannin on endothelial cells.

RESULTS50 nmol/L wortmannin could significantly inhibit the proliferation and migration of endothelial cells induced by HGMCM. The percentage of endothelial cells in S phase was obviously decreased [from (37.82 +/- 0.57)% to (21.91 +/- 0.23)%, P < 0.01], while there was an increase in the percentage of cells in G(0)/G(1) phase [from (54.57 +/- 1.19)% to (65.59 +/- 0.41)%, < 0.01] and G(2)/M phase (< 0.05).

CONCLUSIONWortmannin can inhibit proliferation and migration of endothelial cells induced by HGMCM, suggesting that wortmannin carries an anti-angiogenetic ability in diseased retina.

Androstadienes ; pharmacology ; Animals ; Cell Cycle ; drug effects ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; pharmacology ; Endothelial Cells ; cytology ; Female ; Glucose ; Humans ; Male ; Neovascularization, Pathologic ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; Rabbits ; Retina ; cytology

Objective:To evaluate the clinical, radiological and arthroscopic outcomes after surgical repair for chronic lateral meniscus posterior root (LMPR) avulsion combined with anterior cruciate ligament (ACL) reconstruction.Methods:From July 2015 to June 2017, a total of 33 patients who underwent transtibial pull-out suture repair for chronic LMPR avulsion combined with anatomic single-bundle ACL reconstruction with hamstring graft were retrospectively reviewed. There were 30 males and 3 females with an average age of 27.7±7.5 years (range 17-45 years) and a mean BMI of 25.2±3.7 kg/m 2 (range 19.4-36.7 kg/m 2). All patients were available for at least two years of follow-up. A second-look arthroscopy was performed to evaluate the healing status of the repaired meniscus. Subjective knee function was assessed through Lysholm and Tegner scores. Objective knee stability was evaluated using KT-1000 arthrometer side-to-side difference (SSD) and pivot shift test under anesthesia. The tibiofemoral relationship was evaluated by anterior tibial subluxation (ATS) measured on axial MRI. Between patients with preoperative ATS ≥6 mm (18 patients in the ATS positive group) and <6 mm (15 patients in the ATS negative group), the postoperative ATS and the reduction of ATS was also compared. Results:After a mean follow-up of 27.5±4.0 months (range 24-39 months), the LMPR avulsion completely healed in 23 (70%) cases, partially healed in 9 (27%) cases, failed to heal in 1 (3%) case on second-look arthroscopy. The Lysholm score was increased from 60.4±13.6 to 82.7±11.1 at 1 year and to 91.4±9.1 at 2 years operatively ( F=155.996, P<0.001). The Tegner score was increased from 3(2, 5) to 4(3, 5) at 1 year and 6(4, 6) at 2 years postoperatively (χ 2=47.791, P<0.001). The KT-1000 SSD was decreased from 9.1±3.3 mm to 2.0±1.7 mm ( t=11.197, P<0.001). The result of pivot shift test was also improved (10 grade I, 20 grade II, 3 grade III, preoperatively vs 30 grade 0, 3 grade I, postoperatively, U=5.161, P<0.001). The ATS was reduced from 5.7±3.9 mm to 3.5±3.2 mm ( t=3.530, P=0.001). However, there was no statistically significant decrease in the ATS of the ATS negative group ( t=0.400, P=0.695). The ATS of the ATS positive group was reduced from 8.7±1.8 mm to 5.0±3.3 mm ( t=4.765, P<0.001), and the ATS reduction of the ATS positive group was greater than that of the ATS negative group (3.7±3.3 mm vs 0.3±2.8 mm, t=3.115, P=0.004). Conclusion:In patients undergoing ACL reconstruction, the transtibial pull-out suture repair for chronic LMPR avulsion yielded meniscus healing rate of 97% with improved subjective knee function and objective knee stability and better restored the tibiofemoral relationship for patients with excessive ATS.

OBJECTIVETo explore the relationship between CMV reactivation and KIR haplotype or HLA-Cw genotype in patients after unrelated-donor hematopoietic stem cell transplantation (HSCT).

METHODSFrom January 2003 to December 2008 the HLA-Cw/KIR genotype of 48 patient-donor pairs were determined by polymerase chain reaction with sequence specific primers (PCR-SSP) and sequence specific nucleotide (PCR-SSOP). Posttransplant CMV reactivation was performed by immune histochemically assay.

RESULTSOf 48 patients, 15 were transplanted from unrelated donors with an antigen mismatch for HLA Cw and 33 patient-donor pairs were matched for HLA-Cw. The CMV reaction rate was 66.7% for HLA-Cw mismatch group and 48.5% for HLA-Cw match group (chi(2) = 1.39, P = 0.2375). Thirty-seven donor-patients pairs belonged to group C1 and 11 to group C2, and CMV reaction rate was 64.9% in group C1 and 18.2% in group C2 (chi(2) = 18.13, P < 0.0001). Twenty-six patients received graft from KIR haplotype A (group A donor) and 22 from KIR haplotype B donors (group B donor) and CMV reaction rate was 57.7% in group A donor and 50.0% in group B donor (chi(2) = 0.28, P = 0.5941). The number of donor activating KIRs (aKIRs) was less than that of recipient aKIRs in 34 patient-donor pairs in which the CMV reaction rate was 70.6%, and the number of donor aKIRs was more than that of recipient aKIRs in 14 patient-donor pairs in which the CMV reactivation was 14.3%. There was a significan difference between the two group (chi(2) = 12.44, P = 0.0004).

CONCLUSIONKIR and HLA-Cw genotypes influence the rate of CMV reactivation following non-T cell deleted unrelated donor hematopoietic cell transplantation.

Genotype ; HLA-C Antigens ; genetics ; Haplotypes ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, KIR ; genetics