Huocao(a traditional Chinese herbal medicine) moxibustion is a characteristic technology in Yi medicine suitable for cold-dampness diseases. Huocao, as the moxibustion material, is confusedly used in clinical practice and little is known about its quality control. In this study, UPLC method was used to establish the chemical fingerprint of non-volatile components in Huocao, and the contents of eight phenolic acids such as chlorogenic acid were determined. Multivariate statistical analysis was performed to obtain the indicator components of Huocao for quality evaluation, and thus a comprehensive evaluation system for the quality of Huocao was built. The UPLC fingerprints of 49 batches of Huocao were established, and there were 20 common peaks, of which eight phenolic acids including neochlorogenic acid and chlorogenic acid were identified. Except for three batches of Huocao, the similarity of the other 46 batches was higher than 0.89, suggesting that the established fingerprint method could be used for quality control of the medicinal herb. The correlation coefficient between entropy weight score of the eight phenolic acids and comprehensive fingerprint score in Huocao was 0.875(P<0.01), which indicated that the eight phenolic acids could be used as indicator components for the quality evaluation of Huocao. Furthermore, in multivariate statistical analysis on the common peaks of fingerprint and the contents of the eight phenolic acids, chlorogenic acid, isochlorogenic acid A and isochlorogenic acid C were screened to be the indicator components. The results revealed that the proposed method achieved a simple and accurate quality control of Huocao based on UPLC fingerprint and multi-component content determination, which provided useful data for establishing the quality standard of Huocao.
Chlorogenic Acid ; Entropy ; Hydroxybenzoates ; Quality Control

OBJECTIVE: To observe the clinical efficacy of Tiaohe Yinyang acupotomy (acupotomy for regulating and harmonizing yin and yang) for knee osteoarthritis (KOA). METHODS: A total of 88 patients with KOA were randomized into a acupotomy group and a sham-acupotomy group, 44 cases in each group. In the acupotomy group, acupotomy was applied at yin side (4-5 high stress points i.e. pes anserinus and terminal of popliteus) and yang side (1-2 high stress points i.e. stimulation point of infrapatellar ligament and suprapatellar bursa) of knee joint. In the sham-acupotomy group, sham-acupotomy was applied at the same points as the acupotomy group. The treatment was given once a week for 2 weeks in the two groups. Before and after treatment, the Western Ontario and McMaster Universities arthritis index (WOMAC) score, visual analogue scale (VAS) score, thickness of medial and lateral collateral ligaments of knee joint, motion range of knee joint and plantar pressure distribution were observed in the two groups. In the follow-up of 3 months after treatment, the WOMAC and VAS scores were recorded in the acupotomy group. RESULTS: After treatment, the sub item scores (pain, stiffness and function) and total scores of WOMAC and VAS scores were decreased in the both groups (P<0.05), pain score, function score and total score of WOMAC and VAS score in the acupotomy group were lower than those in the sham-acupotomy group (P<0.05). Before and after treatment, there were no statistical differences in thickness of medial and lateral collateral ligaments of knee joint and motion range of knee joint between the two groups (P>0.05). After treatment, the plantar medial pressure was increased while the plantar lateral pressure was decreased (P<0.05), and the plantar force line moved medially in the acupotomy group. In the follow-up, the sub item scores and total score of WOMAC and VAS score were lower than those before and after treatment in the acupotomy group (P<0.05). CONCLUSION Tiaohe Yinyang acupotomy can improve the clinical symptoms of knee joint in patients with KOA by changing the local biological stress.
Humans ; Social Group ; Pain

ObjectiveUsing Chromium-51 release assay, lactate dehydrogenase release assayand other methods to detect the cytotoxicity of CD19 CAR-T cells is cumbersome, with low repeatability and poor stability. This study aims to establish a label-free and real-time method for detectingspecific cytotoxicity of CD19 CAR-T cells.MethodsIn order to establish target cell models for cytotoxic assay of CD19 CAR-T cells by using Real Time Cellular Analysis (RTCA) system,the adherent human breast cancer cells were infected with lentiviral vectors encoding CD19. CD19 expression on the transduced cells was detected by flow cytometry. The cellsexpressing CD19 stably werethen sorted by fluorescence activated cell sorting (FACS).With such cells as target cells, CD19 CAR-T cells and BCMA CAR-T cells as effector cells, RTCAsystem was used to evaluate the cytotoxicity of CAR-T cells against target cells.ResultsMDA-MB-231 and SKBR3cells with stable expression CD19were obtained in this study.The results of flow cytometry showed that positive expression rate of CD19 in MDA-MB-231/CD19 cells and SKBR3/CD19 monoclonal cells were 99.03% and 98.91%,respectively.RTCA results showed that with MDA-MB-231 and MDA-MB-231/CD19 cells as target cells,CD19 CAR-T cells showed significant cytotoxicity to MDA-MB-231/CD19 cellsat the effector-target ratio of 5∶1, 1∶1 and 1∶5,but not to MDA-MB-231 cells. With SKBR3 and SKBR3/CD19 cells as target cells, CD19 CAR-T cells showed significant cytotoxicity to SKBR3/CD19 cellsat the effector-target ratio of 5∶1and 1∶1. When the effector-target ratio was 1∶5, there was no obvious cytotoxicity.The data of MDA-MB-231/CD19 or SKBR3/CD19 as target cells and CD19 CAR-T as effector cells were analyzed separately, showing that when the number of target cells was the same, the cytotoxicity detected by RTCA increased as the number of CD19 CAR-T cells increased.The cytotoxic assays of CD19 CAR-T cells showed specificity and dose-response relationship of CD19 CAR-T cytotoxicity against the target cells.ConclusionThis study established a method for evaluating cytotoxicity of CD19 CAR-T cells that is real-time, label-free, simple and convenient.

OBJECTIVE: To estimate the univariate heritability of resting heart rate and common chronic disease such as hypertension, diabetes, and dyslipidemia based on extended pedigrees in Fujian Tulou area and to explore bivariate heritability to test for the genetic correlation between resting heart rate and other relative phenotypes. METHODS: The study was conducted in Tulou area of Nanjing County, Fujian Province from August 2015 to December 2017. The participants were residents with Zhang surname and their relatives from Taxia Village, Qujiang Village, and Nanou Village or residents with Chen surname and their relatives from Caoban Village, Tumei Village, and Beiling Village. The baseline survey recruited 1 563 family members from 452 extended pedigrees. The pedigree reconstruction was based on the family information registration and the genealogy booklet. Univariate and bivariate heritability was estimated using variance component models for continuous variables, and susceptibility-threshold model for binary variables. RESULTS: The pedigree reconstruction identified 1 seven-generation pedigree, 2 five-generation pedigrees, 23 four-generation pedigrees, 186 three-generation pedigrees, and 240 two-generation pedigrees. The mean age of the participants was 57.2 years and the males accounted for 39.4%. The prevalence of hypertension, diabetes, dyslipidemia in this population was 49.2%, 10.0%, and 45.2%, respectively. The univariate heritability estimation of resting heart rate, hypertension, and dyslipidemia was 0.263 (95%CI: 0.120－0.407), 0.404 (95%CI: 0.135－0.673), and 0.799 (95%CI: 0.590－1), respectively. The heritability of systolic blood pressure, diastolic blood pressure, fasting glucose, total cholesterol, triglyceride, high-density lipoprotein cholesterol, and low-density lipoprotein cholesterol was 0.379, 0.306, 0.393, 0.452, 0.568, 0.852, and 0.387, respectively. In bivariate analysis, there were phenotypic correlations between resting heart rate with hypertension, diabetes, diastolic blood pressure, fasting glucose, and triglyceride. After taking resting heart rate into account, there were strong genetic correlations between resting heart rate with fasting glucose (genetic correlation 0.485, 95%CI: 0.120－1, P<0.05) and diabetes (genetic correlation 0.795, 95%CI: 0.181－0.788, P<0.05). CONCLUSION Resting heart rate was a heritable trait and correlated with several common chronic diseases and related traits. There was strong genetic correlation between resting heart rate with fasting glucose and diabetes, suggesting that they may share common genetic risk factors.
Blood Pressure ; Chronic Disease ; Female ; Heart Rate ; Humans ; Hypertension ; Male ; Middle Aged ; Pedigree

Objective To compare the efficacy and complications of percutaneous nephroscope decortication of cystic renal disease with transureteroscopic decompression of cystic renal disease.Methods Retrospectively analyze the clinical data of 42 simple renal cyst cases,who under treatment of surgical in Zhongnan Hospital of Wuhan University from Sep.2010 to Sep.2014 via percutaneous nephrolithotomy as well as ureteroscope.There were 21 patients in each group.Comparisons were made between the two groups on operation time,peripheral tissue injury,blood loss,postoperative infection,hospitalzation time.Postoperative recurrence were followed up.Results For the percutaneous nephroscope decortication of cystic renal disease group and transureteroscopic decompression of cystic renal disease group,the operation time were (38.43 ± 9.76) minutes,(28.95 ± 8.67) minutes,the number of tissue injury were 8,6;the blood loss were (28.62 ± 9.82) mL,(23.48 ± 7.65) mL;the number of postoperative infection was 4,10;the postoperative recurrence were 2,5;the hospitalzation time were 2 days and 8 days.Compared with the transureteroscopic decompression group,the percutaneous nephroscope decortication group had a less postoperative infection and fewer postoperative recurrence (P ＜ 0.05).But the operation time was longer in the percutaneous nephroscope decortication group (P ＜ 0.05).Conclusions The therapeutic effect of percutaneous nephroscope decortication is much better than that of transureteroscopic decompression,but also has a little disadvantage.

OBJECTIVETo study the changes in the mRNA expression of insulin-like factor 3 (INSL-3) in the testis of mouse models of flutamide-induced cryptorchidism.

METHODSWe randomized pregnant BALB/c mice to groups A (control) , B, C, D and E to receive continuous gavage of flutamide at 0, 150, 300, 500 and 700 mg/kg body weight, respectively, from gestation day 12 to 21. We detected the expression of INSL-3 mRNA in the testis of the neonates by real-time PCR at 4 and 8 postnatal weeks.

RESULTSNo cryptorchidism was found in group A; unilateral cryptorchidism was seen in groups B (10.0%) and C (25.0%); and bilateral cryptorchidism was observed in groups D (21.1%) and E (40.0%). The expression of INSL-3 mRNA was reduced with the increased dose of flutamide, not significantly changed in groups B and C (P > 0.05) but remarkably decreased in D and E as compared with A (P < 0.05).

CONCLUSIONAdministration of flutamide to pregnant mice can induce unilateral cryptorchidism at 150 and 300 mg/kg and bilateral cryptorchidism at 500 and 700 mg/kg in their male offspring, which may be related with its reducing effect on the expression of INSL-3 in the testis of the mice.

Animals ; Cryptorchidism ; chemically induced ; metabolism ; Female ; Flutamide ; toxicity ; Insulin ; metabolism ; Male ; Maternal Exposure ; adverse effects ; Mice ; Mice, Inbred BALB C ; Pregnancy ; Proteins ; metabolism ; RNA, Messenger ; genetics ; Testis ; metabolism

OBJECTIVETo study the expression and the role of heme oxygenase-1 (HO-1) and inducible nitric oxide synthase (iNOS) in preterm rats with hyperoxia-induced lung injuries.

METHODSSixty-four three-day-old preterm Sprague-Dawley rats were randomly assigned to a hyperoxia group (90% oxygen exposure) and a control group (room air exposure), with 32 rats in each group. After 3 days or 7 days of exposure, the lung activity of HO-1 and nitric oxide (NO) contents in bronchoalveolar lavage fluid (BALF), pulmonary histopathologic changes, and the cellular distribution and expression of HO-1 and iNOS in the lungs were measured.

RESULTSAfter 3 days and 7 days of exposure, the hyperoxia group showed acute lung injuries characterized by the presence of hyperaemia, red cell extravasation and inflammatory infiltration. The NO contents in BALF and the iNOS expression in the lungs increased significantly in the hyperoxia group compared with those in the control group 3 and 7 days after exposure. The expression of HO-1 in macrophages in the lungs increased significantly in the hyperoxia group compared with that in the control group 3 and 7 days after exposure. The NO contents in BALF and the iNOS and HO-1 expression in the lungs increased significantly 7 days after hyperoxia exposure compared with 3 days after hyperoxia exposure.

CONCLUSIONSiNOS and HO-1 levels in the lungs increase in preterm rats with hyperoxia-induced lung injuries, suggesting that iNOS and HO-1 may play roles in hyperoxia-induced pulmonary injuries.

Animals ; Bronchoalveolar Lavage Fluid ; chemistry ; Female ; Heme Oxygenase (Decyclizing) ; analysis ; physiology ; Hyperoxia ; complications ; enzymology ; Lung ; enzymology ; Lung Injury ; etiology ; Male ; Nitric Oxide Synthase Type II ; analysis ; physiology ; Rats ; Rats, Sprague-Dawley

OBJECTIVEHemophilia A is an inherited bleeding disorder caused by defects in factor VIII (FVIII) gene. In the present study, the frequencies of the microsatellite alleles at introns 13 and 22 in the factor VIII gene were analyzed in the group of Han nationality in Guangxi Zhuang Autonomous Region to explore their diagnostic value for hemophilia A. These two sites were also used to detect the carriers in 13 hemophilia A families.

METHODSNinty-one individuals of Han ethnic group in Guangxi Zhuang Autonomous Region (135 X chromosomes) and 13 HA families were subjected to molecular studies. First, these two fragments were PCR amplified simultaneously. Then, silver staining was used later to show their polymorphisms. The investigators selected one sample at random to obtain its lengths of the PCR products at these two sites by ABI310 PCR amplifier. After counting its repeated numbers of (CA) according to the documents concerned, the repeated numbers of the other samples could be counted easily.

RESULTSIn the 91 individuals, 6 and 4 alleles were detected at these two sites, respectively. At intron 13 the allele frequencies ranged from 0.0002 to 0.5408 and polymorphism information content (PIC) was 0.5899. At intron 22 the allele frequencies ranged from 0.0444 to 0.4963 and its PIC was 0.5359. The actual heterozygosity for intron 13 and intron 22 were 0.6364 (28/44) and 0.5227 (23/44), respectively. In 13 hemophilia A families with positive history, 9 of them were diagnosed by this method and the diagnosis rate was 69%.

CONCLUSIONWith high PICs, (CA)n at intron 13 and intron 22 were two valuable sites in the diagnosis of hemophilia A in the population of Han ethnic group in Guangxi Zhuang Autonomous Region. Compared with some other HA restrictive fragment length polymorphisms (RFLP), intron 22 (GT)n (AG)n was more informative.

Alleles ; Amplified Fragment Length Polymorphism Analysis ; Asian Continental Ancestry Group ; genetics ; Factor VIII ; genetics ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Hemophilia A ; diagnosis ; genetics ; Heterozygote ; Humans ; Introns ; Male ; Polymerase Chain Reaction ; Polymorphism, Genetic ; Sequence Analysis, DNA ; Silver Staining

Objective To investigate whether leukotriene D4 (LTD4) regulates eotaxin-3 (Eot-3) expression in bronchial epithelial cells, and study effect of pranlukst on the regulation.Methods BEAS-2B cells and normal human bronchial epithelia cells were pre- treated with LTD4 for 1 hour,stimulated with interleukin-4, the cells were incubated for 24 hours. Eot-3 protein in supernatant were measured by enzyme linked immunosorbent assay(ELISA). The cells were pretreated with pranlukast in different concentration, then the above procedure was repeated. Results The untreated bronchial epithelial cell expressed Eot-3 protein on a very low level. After stimulating with IL-4 and incubating for 24 hours, Eot-3 production increased significantly. Pretreating the cells with LTD4 enhanced the inducing effect of IL-4. Pranlukast inverted the upregulation of LTD4. Conclusions Upregulating the expression of Eot-3 induced by IL-4 on bronchial epithelial cells may explain partially the mechanism of leukotrienes involving airway allergic inflammation of asthma. The invertion impact on upregulation of LTD4 by pranlukast may be one of mechanisms that leukotrienes receptor antagonist cure asthma.

Objective To explore the effect of protoporphyrin Ⅸ zinc(Znpp) on hyperoxic lung injury in preterm rats.Methods Three-day-old preterm SD rats were randomly assigned to room air control group(group Ⅰ)hyperoxia control group(oxygen≥900 mL/L)(group Ⅱ),room air plus Znpp group(group Ⅲ),hyperoxia plus Znpp group(group Ⅳ).Group Ⅲ and group Ⅳ were injected intraperitoneally with ZnPP 45 ?mol/kg each day.After the third day and the 7th day of exposure,the activity of heme oxygenase-1(HO-1) and the percent of carboxyhemoglobin(HbCO) in the lungs,the lung wet weight /dry weight ratio(W/D),tumor necrosis factor-?(TNF-?),total protein and malondialdehyde(MDA) in bronchoalveolar lavage fluid were determined and lung histophathological changes were examined in all groups.Results On the third day,compared with group Ⅰ the activity of HO-1 and the percent of HbCO in the lungs,W/D,TNF-?,total protein and MDA,all greatly rised in group Ⅱ(Pa