Development of the method for determinating ADCC biological activity of IVIG / 中国输血杂志

【Objective】 To establish a method for determinating the antigen-dependent cell-mediated cytotoxicity (ADCC) of human immunoglobulin (pH4)for intravenous injection (IVIG) on luciferase reporter gene-modified cell assay. 【Methods】 As effector cells, Jurkat-NFAT-Luc-CD16 cells were used in the assay, and PLC/PRF/5 cells were used as target cells. After incubation of effector cells and target cells with IVIG, the method for determinating ADCC biological activity of IVIG was established by detecting luciferase released by activated T nuclear factor after binding of IVIG Fc fragment to effector cells. Meanwhile, the experimental assay conditions were optimized, and the methodology was verified subsequently. 【Results】 IVIG had a dose-response relationship in this method, which was consistent with four parameter logistic model. And the PLC/PRF/5 cells were finally determined as the target cells. The initial dilution concentration of antibody was 20 mg/mL, and the ratio dilution was 1∶2, and the effector to target ratio was 1∶3, and co-incubation time of two cells and IVIG was 24 hours. Within-run and between-run analysis including three independent tests, initial working concentration relative light unit (RLU) and the relative standard deviation (RSD) of the concentration for 50% of maximal effect(EC50) were less than 11%. The relative titers of the recovery samples of the two different dilution groups were (23.50±1.69)% and (49.30±2.97)%, respectively, and the corresponding recovery rates were (93.50±6.30)% and (96.24±5.43)%, respectively, with RSD less than 11%. 【Conclusion】 The method for determinating ADCC biological activity of IVIG based on luciferase reporter gene-modified cell assay was successfully established. It could be applied in determinating the ADCC biological activity of IVIG, and has the advantages of satisfactory linearity, accuracy, precision and specificity.

Effect of IgG fragments in IVIG(pH4)on phagocytosis of sensitized erythrocytes by macrophage / 中国输血杂志

【Objective】 To research the effect of the Fc, Fab and F(ab′)2 fragments of immunoglobulin G, the main components of Human Immunoglobulin(pH4) for Intravenous Injection(IVIG), on the phagocytic function of macrophages derived from THP-1 cells. 【Methods】 First of all, IVIG was digested with papain and pepsin to obtain Fc, Fab and F(ab′)2, and these components were then identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Afterwards, propylene glycol monomethyl ether acetate (PMA) was used to induce THP-1 cells to differentiate into M0 macrophages. Finally, the sensitized erythrocytes were labeled with carboxy fluorescein succinimidyl ester (CFSE), and the effect of the above components on the phagocytic ability of M0 macrophages to engulf sensitized erythrocytes was detected by flow cytometry. 【Results】 The identification results of SDS-PAGE showed that the prepared IgG fragments met the requirements of subsequent experiments. Flow cytometry performs showed that the phagocytosis model of M0 macrophages had been successfully established. When the concentration of Fc increased from 0.1μg/ mL to 10μg/ mL, the phagocytosis rate of erythrocytes sensitized by M0 macrophages decreased from (24.21±0.58) % to (12.27±0.19) %. When the concentration of IVIG protein increased from 0.1 μg/ml to 10 μg/ml, the phagocytosis rate decreased from (20.57±0.39) % to (0.20±0.03) %. Meanwhile, at the same protein concentration (10 μg/ml), the inhibitory effect of Fc on phagocytosis was only half that of IVIG. In addition, Fab, F(ab′)2, and human serum albumin could not inhibit phagocytosis of M0 macrophages. 【Conclusion】 IVIG can effectively inhibit the phagocytosis of THP-1 derived M0 macrophages, which is mainly dependent on the Fc, but not related to the Fab of IgG and F (ab′)2.

Maturation of Stem Cell-Derived Cardiomyocytes: Foe in Translation Medicine

With the in-depth study of heart development, many human cardiomyocytes (CMs) have been generated in a laboratory environment. CMs derived from pluripotent stem cells (PSCs) have been widely used for a series of applications such as laboratory studies, drug toxicology screening, cardiac disease models, and as an unlimited resource for cell-based cardiac regeneration therapy. However, the low maturity of the induced CMs significantly impedes their applicability. Scientists have been committed to improving the maturation of CMs to achieve the purpose of heart regeneration in the past decades. In this review, we take CMs maturation as the main object of discussion, describe the characteristics of CMs maturation, summarize the key regulatory mechanism of regulating maturation and address the approaches to promote CMs maturation. The maturation of CM is gradually improving due to the incorporation of advanced technologies and is expected to continue.

Effect of residual sex hormones in sex-differentiated human IgG samples on DC secretion of inflammation-related factors / 中国输血杂志

【Objective】 Tostudy the effect of sex-differentiated human IgG samples on Dendritic cells (DC) secretion of inflammation-related factors and explore the effect of residual sex hormones in IgG products (such as IVIg) on the secretion of IL-6 by DC. 【Methods】 According to the standard IVIg production process, the company was entrustedto prepare sex-differentiated plasma purified IgG samples, and two sex-differentiated IgG samples with different sex ratios (male to female ratio1: 0, 0: 1) were obtained. The samples and referenceswere treated with human DC (induced by THP-1 cells) respectively. After 24 h of culture, the chemokines (CCL2, CCL3, CCL4), adhesion molecule (ICAM-1) and inflammatory cytokines (IL-1, IL-6, IL-10, IL-12p70, IFN-) in the cell supernatant were detected, The effects of different samples on the secretion of inflammation-related factors by DC were compared. The effect of sex hormone residues on the anti-inflammatory ability of IgG products was preliminarily explored uing sex hormones and sex hormone receptor blockers. 【Results】 The samples in each group significantly inhibitedthe secretion of chemokines (CCL2, CCL3, CCL4) and the adhesion molecule (ICAM-1) by mature DC (compared with the PBS group, P<0.05), but significantly promoted the secretion of inflammatory cytokines (IL-1a/b, IL-6, IL-10, IL-12p70), compared with the PBS group(P<0.05). The results of sex hormone residues showed that therewere residues of estradiol (E2) and testosterone (TSTO) in sex-differentiated IgG samples and IVIg products. The experimental results of IVIg and sex hormone/sex hormone receptor blockers showed that residual E2 may promote the secretion of IL-6 by DC, which may be achieved through the E2 receptor ERb. 【Conclusion】 There are differences in the effect of IgG samples prepared from combined plasma with different sex ratios on the secretion of cytokines by DC, which may be related to the residual E2 in the products. The residual sex hormones in IVIg may promote the production and secretion of IL-6 through the sex hormone receptor ERb expressed in DC, and TSTO may have a collaboration effect to enhance the secretion-promoting effect of IL-6 by E2. This study provides a theoretical basis for whether sex hormone residues need to be considered in the quality control indicators of IVIg products.

Effect of glutamine metabolism on chemoresistance and its mechanism in tumors / 浙江大学学报·医学版

The metabolic reprogramming of tumor cells is characterized by increased uptake of various nutrients including glutamine. Glutamine metabolism provides the required substances for glycolysis and oxidative phosphorylation and affects the homeostasis of carbohydrate，fat and protein metabolism to induce the chemoresistance of tumor cells. Combination of chemotherapeutic agents with inhibitors specific to different components of glutamine metabolic pathway has obtained favorable clinical results on various tumors. Glutamine metabolic pathway plays a role in drug resistance of tumor cells in various ways. Firstly，the dynamic change of glutamine transporters can directly affect intracellular glutamine content thereby causing drug resistance; secondly，tumor stromal cells including adipocyte，fibroblast and metabolite from tumor microenvironment would give rise to immune-mediated drug resistance; thirdly，the expression and activity of key enzymes in glutamine metabolism also has a critical role in drug resistance of tumors. This article reviews the effects of glutamine metabolic pathway in the development of tumor chemoresistance，in terms of transporters，tumor microenvironment and metabolic enzymes，to provide insight for improving the therapeutic efficacy for drug-resistant tumors.

Determination of the Concentration of Sinoacutine and Its Pharmacokinetic Study in Rabbits / 中国药房

OBJECTIVE： To establish a method for concentration determination of sinoacutine in rabbit plasma， and to conduct its pharmacokinetic study. METHODS： The rabbits were grouped according to gender， 6 rabbits in each group. Rabbits were injected with sinoacutine solution （5 mg/kg） via ear vein. Each blood sample 1 mL was collected before medication and 5， 10， 15， 30， 45， 60， 90， 120， 180， 240 min after medcation. After the plasma isolated and extracted with ethyl acetate， HPLC method was adopted by using sinomenine as internal standard. The determination was performed on Agilent Zorbax Extend-C18 column with mobile phase consisted of methanol-2 mmol/L disodium hydrogen phosphate aqueous solution （containing 0.016％ triethylamine， pH 9.8） （45 ∶ 55， V/V) at the flow rate of 1 mL/min. The detection wavelength was set at 262 nm， and column temperature was 30 ℃. The sample size was 20 μL. The pharmacokinetic parameters were calculated by using DAS 3.0 software. The difference of 2 groups were investigated by t-test. RESULTS： The linear range of sinoacutine were 0.1-5.0 mg/L； the limit of quantitation was 0.1 mg/L， and the lowest detection limit was 0.08 mg/L. RSDs of intra-day and inter-day were both less than 10％； the accuracy ranged from （99.80±8.21）％-（103.61±8.55）％. The extraction method did not affect the quantitative analysis of the substance to be measured. The average plasma-time curve of sinomenine with single intravenous injection in rabbits was in line with the two-compartment model. The distribution half-life of all rabbits was （10.99±2.52） min， and the elimination half-life was （147.08±32.41） min. AUC0-t was （190.82±30.82）mg·min/L， and AUC0-∞ was （289.82±73.27） mg·min/L. There was no statistical significance in pharmacokinetic parameters between female and male rabbits （P＞0.05）. CONCLUSIONS： Established HPLC method is simple， specific and sensitive， and can be used for plasma content determination of sinoacutine. Pharmacokinetic study shows that the pharmacokinetic process of the compound is in line with two-compartment model in rabbits. The pharmacokinetic parameters of the compound have no sex difference， and the compound is distributed rapidly and eliminated fast.

Clinical analysis of 95 cases of cervical tuberculous lymphadenopathy associated with abscess formation / 中国感染与化疗杂志

Objective To explore the clinical features and surgical treatment of cervical tuberculous lymphadenopathy associated with abscess formation.Methods The clinical data of 95 cases of cervical tuberculous lymphadenopathy associated with abscess formation were reviewed and analyzed retrospectively.All the patients were treated in Shenzhen Third People's Hospital during the period from February 2012 to May 2014.Results The mean age of the 95 patients was (27.8±8.2) years.The average duration of the disease was 4.0 months.The tuberculous lymphadenopathy was most frequently found in Level Ⅳ cervical lymph nodes.Thirty-four patients were complicated with pulmonary tuberculosis.History of surgical treatment for tuberculous cervical lymphadenopathy was reported in 26 cases.Sixty-five patients were managed with incision and drainage plus dress changing.Twenty-two patients underwent abscess rinsing,excision and primary wound closure.And 8 patients received excision of ulcers and sinuses,and lymphadenopathy cleaning.The average healing time was (2.11± 1.76) months for incision and drainage.Abscess and ulcer reoccurred in 18 of the 65 patients after healing completely by incision and drainage.All the patients treated by the other two surgical modalities were cured as primary healing except 2 patients,who received local flap grafting after primary surgery.Conclusions Cervical tuberculous lymphadenopathy associated with abscess formation has special features in diagnosis and treatment.Surgical approach is usually effective if optimized for individual patients.

Analysis of risk factors of metabolic syndrome in patient with rheumatoid arthritis / 中华风湿病学杂志

Objective To explore the relationships between metabolic syndrome (MS) and rheumatoid arthritis (RA).Methods Two hundred and forty RA patients who fulfilled the 1987 revised American College of Rheumatology (ACR) classification criteria,and 250 age and sex matched healthy controls were enrolled.The frequency of metabolic syndrome was assessed using three Metabolic Syndrome definitions (Guidelines on Prevention and Treatment of Blood Lipid Abnormality in Chinese Adults 2016,International Diabetes Federation 2005,National Cholesterol Education Program 2005).Data were analyzed by Chi-square test and t test,risk factors were identified by Logistic regression.Results ① According to the definition used,the frequency of metabolic syndrome varied from 28.3％ to 35％ in RA,which was significantly higher than controls.② In RA with MS group defined by guidelines on Prevention and Treatment of Blood Lipid Abnormality in Chinese Adults 2016,the age,the diseases duration,erythrocyte sedimentation rate (ESR),DAS28,health assessment questionnaire (HAQ) scores and dose of glucocorticoid used were higher than controls [(12±6) years vs (10±7) years,t=6.96,P=0.01,(32±9) mm/1 h vs (26±6) mm/1 h,t=4.66,P=0.008,(3.7±1.2) vs (2.6±1.0),t=3.78,P=0.01,(1.6±0.5) vs (1.2±0.5),t=3.42,P=0.017],the percentage of patients who received anti-tumor necrosis factor α therapy was lower than the control group (39.7％ vs 23.7％,x2=6.881,P=0.009).③ In multivariate analysis,long disease duration,high ESR and HAQ scores were independently associated with the presence of MS [OR(95％CI):1.489(0.382,0.624),1.555(0.423,0.729),1.603(0.423,0.858),P＜0.01].Being treated with anti-tumor necrosis factor α was an independent protector of the presence of metabolic syndrome in RA patients [OR=0.582,95％CI(0.453,0.742),P＜0.01].Conclusion The frequency of metabolic syndrome in RA is higher compared to controls,while diseases duration,ESR,and HAQ are independent predictors for the presence of metabolic syndrome in patients with RA,anti-TNF-α therapy may be protective factor for the presence of metabolic syndrome in patients with RA.

The effects of exposure to the famine during early life with elevated resting heart rate in the adult / 中华预防医学杂志

<p><b>OBJECTIVE</b>To investigate the association between exposure to the famine during early life and elevated resting heart rate (RHR) in adulthood.</p><p><b>METHOD</b>From June 2006 to October 2007, the employees of kailuan group who took part in the health examination were selected. Of those, 18 619 cases who was born during October 1, 1956 to September 30, 1964 in Hebei province were finally included in the analysis based on the inclusion and exclusion criteria. All the subjects were received questionnaire survey, smoking and drinking, physical examination, Lab examination and the measurement of RHR. The subjects of famine exposure group (3 190 cases) were born from October 1, 1959 to September 30, 1961, semi-exposure group (3 851 cases) were born from October 1, 1958 to September 30, 1959 and from October 1, 1961 to September 30, 1962, control group (11 578 cases) were born from October 1, 1956 to September 30, 1958 and from October 1, 1962 to September 30, 1964. The RHR and the detection rate of elevated RHR were compared among the three groups. The Multivariate logistic regression model was used to analyze the association between of exposure to famine during early life and elevated RHR in adulthood.</p><p><b>RESULTS</b>The RHR level was higher in famine exposure group and semi-exposed group than control group, which were (74.34 ± 9.71), (74.41 ± 9.48) and (73.90 ± 9.45) beat per minute (bpm) (P values were 0.003 and 0.020, respectively). In all of the subjects. The results of multivariate logistic regression showed that exposure of famine during early life increased the risk of elevated RHR in adulthood after adjustment for age, gender and other confounders (OR = 1.10, 95% CI: 1.01-1.21). In men, exposure of famine during early life also increased the risk of elevated RHR in adulthood (OR = 1.15, 95% CI: 1.04-1.28); In women, there was no association between the famine exposure and elevated RHR (OR = 0.92, 95% CI: 0.74-1.14).</p><p><b>CONCLUSION</b>Exposure of famine during early life increases the risk of elevated RHR in adulthood. This negative effect existed mainly in the male.</p>

Effects related to experiences of famine during early life on diabetes mellitus and impaired fasting glucose during adulthood / 中华流行病学杂志

<p><b>OBJECTIVE</b>To investigate the effects of famine-experience during early life on diabetes mellitus and impaired fasting glucose in the adulthood.</p><p><b>METHODS</b>In a total of 101 510 employees who took part in the health examination at the Kailuan Group between 2006 to 2007 were recruited. All the study subjects were born in Hebei province between 1956-10-01 and 1964-09-30 but those who had incomplete data were excluded. 19 347 subjects were finally included for analysis. Members from the famine-exposed group were born between Oct. 1, 1959 and Sep. 30, 1961. There was a semi-exposed group with members born between Oct. 1, 1958 and Sept. 30, 1959 and from Oct. 1, 1961 to Sept. 30, 1962 but members from the control group were born from Oct. 1, 1956 to Sept. 30, 1958 and from Oct. 1, 1962 to Sept. 30, 1964. Prevalence rates on diabetes mellitus and the detection rate of impaired fasting glucose among the three groups were compared. Logistic regression model was used to analyze the effects of famine-experience during early life with the prevalence of diabetes mellitus and the detection rate of impaired fasting glucose during adulthood.</p><p><b>RESULTS</b>Prevalence of diabetes mellitus and the detection rate of impaired fasting glucose in the famine-exposed adult-cohort groups were 8.99%, 8.96% while 8.05% and 9.35% in the semi-exposure groups, 7.71% and 8.20% in the control group. Results from the multivariate logistic regression analysis showed that experiences of famine during early life increased the risk of diabetes mellitus and impaired fasting glucose in adulthood with the odds ratios as 1.218 (95% CI: 1.056-1.404, P = 0.007) and 1.142 (95% CI: 0.994-1.312, P = 0.061). After stratification by sex, odds ratios in males were 1.163 (95% CI: 1.001-1.350, P = 0.048)and 1.213(95% CI:1.039-1.417, P = 0.015). The odds ratios in females were 1.319 (95% CI: 0.920-1.891, P = 0.132) and 0.990 (95% CI: 0.679-1.444, P = 0.959).</p><p><b>CONCLUSION</b>Experiences of famine during early life increased the risk of diabetes mellitus and impaired fasting glucose in the adulthood. However, this negative effect existed mainly in the males, according to the results from our study.</p>

Nuclear matrix associated protein PML: an arsenic trioxide apoptosis therapeutic target protein in HepG2 cells / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate arsenic trioxide (As(2)O(3))-induced apoptosis and the effects on cell nuclear matrix related protein promyelocytic leukaemia (PML).</p><p><b>METHODS</b>HepG2 cells were cultured in MEM medium and treated with 0.5, 2, 5 and 10 micro mol/L As(2)O(3) for either 24 h or 96 h at each concentration. In situ terminal deoxynucleotidyl transferase (TdT) labeling (TUNEL) and DNA ladders were used to detect apoptosis. Confocal microscopy and Western blotting were used to observe the expression of PML.</p><p><b>RESULTS</b>The growth rates of HepG2 cells were slower in the As(2)O(3) treated than the untreated control group. DNA ladder and TUNEL positive apoptotic cells could be detected in As(2)O(3) treated groups. The expression of PML decreased in HepG2 cells with 2 micro mol/L As(2)O(3) treatment. Confocal images demonstrated that the expression of PML protein in HepG2 cell nuclei decreased after treatment with 2 micro mol/L As(2)O(3), and micropunctates characteristic of PML protein in HepG2 cell nuclei disappeared after treatment with 5 micro mol/L As(2)O(3).</p><p><b>CONCLUSIONS</b>Our results show that arsenic trioxide can significantly inhibit the growth of HepG2 cells in vitro. As(2)O(3) induces apoptosis in HepG2 tumor cells in a time and concentration dependent manner. As(2)O(3) may degrade the PML protein in HepG2 cell nuclei. The decreased expression of PML in As(2)O(3) treated tumor cells is most likely to be caused by apoptosis. Nuclear matrix associated protein PML could be the target of As(2)O(3) therapy.</p>

The Effect of Compound Glycyrrhizin on Jaundice after Cholelithes Operation / 中国药房

OBJECTIVE:To observe the effect of compound glycyrrhizin on patients with cholelithes combined with ob?structive jaundice.METHODS:The changes of bile drainage and liver function in41patients with choledocholith and obstruc?tive jaundice were observed on1to15days after operation(the trial group).The control group did not receive compound gly?cyrrhizin.RESULTS:The bile drainage of the trial group arrived to stable state3days earlier than that of the control group and color of bile was darker than that of control group;The liver function of trial group was significantly better than that of the control group from the7th day after operation(P

Immunohistochemical study on STAT_3 change after focal cerebral ischemic reperfusive injury in rats / 临床神经病学杂志

Objective To explore the expression of singal transducers and activators of transcription(STAT 3) during focal cerebral ischemic reperfusive injury in rats and the relationship between ischemic neuronal damage and it.Methods Using immunohistochemical method of avidinbiotin peroxidase complex (ABC) we observed the distribution of positive cells in STAT 3 protein immunoreaction after focal cerebral ischemic reperfusive injury in rats.Results STAT 3 immunoreactive positive cells were not found in the cortex and striatum of normal and sham operative rat brains and nonischemic hemisphere brain after cerebral ischemia,small amount of STAT 3 immunity positive cells were induced in the embolism la teral infarction area 12 h after reperfusive injury,and peaked after 24 h,especially in ischemic lateral striatum and around cortex,small number of nerve cells in around infarction still showed positive expression after one week.The difference had remarkable significance( P

Breast-conserving modified radical mastectomy for breast cancer:a report of 21 cases / 中国普通外科杂志

Objective To explore the indication,surgical procedure and the postoperative treatment of breast conserving modified radical mastectomy for early stage breast cancer .Methods The clinical data of 21 patients with early stage breast cancer underwent breast conserving modified radical mastectomy were analyzed retrospectively. Results All patient recovered uneventfully. All the external configuration of the breast were fine. There were no recurrence and no complications in this series. Conclusions The breast conserving modified radical mastectomy is recommended for the early stage breast cancer. The external configuration of breast is fine postoperatively, and patients have higher survival quality.But follow up is necessary in order to find and treat the recurrence of breast cancer.