Polysaccharides from Chinese herbal medicine: a review on the hepatoprotective and molecular mechanism / 中国天然药物

Polysaccharides, predominantly extracted from traditional Chinese medicinal herbs such as Lycium barbarum, Angelica sinensis, Astragalus membranaceus, Dendrobium officinale, Ganoderma lucidum, and Poria cocos, represent principal bioactive constituents extensively utilized in Chinese medicine. These compounds have demonstrated significant anti-inflammatory capabilities, especially anti-liver injury activities, while exhibiting minimal adverse effects. This review summarized recent studies to elucidate the hepatoprotective efficacy and underlying molecular mechanisms of these herbal polysaccharides. It underscored the role of these polysaccharides in regulating hepatic function, enhancing immunological responses, and improving antioxidant capacities, thus contributing to the attenuation of hepatocyte apoptosis and liver protection. Analyses of molecular pathways in these studies revealed the intricate and indispensable functions of traditional Chinese herbal polysaccharides in liver injury management. Therefore, this review provides a thorough examination of the hepatoprotective attributes and molecular mechanisms of these medicinal polysaccharides, thereby offering valuable insights for the advancement of polysaccharide-based therapeutic research and their potential clinical applications in liver disease treatment.

Pharmacokinetics of antibody-drug conjugates and their research progress in the treatment of gastrointestinal malignancies / 药学学报

The morbidity and mortality of gastrointestinal malignancies are the highest in the world. For patients with poor response to conventional chemotherapy, new treatment methods are urgently needed. In recent years, under the background of precision medicine, antibody-drug conjugates (ADCs) with high tumor specificity and potent toxicity have become a hot research spot in the field of biomedicine. However, due to the complex structure and mechanism of ADCs, its pharmacokinetic research is facing great challenges which are the biggest resistance to the development of ADCs at present. In this case, it is of great significance to understand the pharmacokinetic properties of ADCs and make use of it to improve the efficacy of ADCs in the treatment of gastrointestinal malignancies. Based on the basic composition and mechanism of ADCs, this review summarizes the pharmacokinetic properties of ADCs, discusses its recent advances in the treatment of gastrointestinal malignancies, in order to provide more references for follow-up research on ADCs.

Protective effect of chrysin regulates AMPK-NLRP3 signaling mediated pyroptosis to alleviate hepatic fibrosis / 药学学报

This study investigated the protective effect of chrysin on hepatic fibrosis by regulating AMP-activated kinase (AMPK)-NOD-like receptor protein 3 (NLRP3) mediated pyroptosis pathway. The hepatic fibrosis model of mice was established by thioacetamide (TAA) in vivo. Except the control and chrysin alone groups, the mice were injected intraperitoneally with TAA at 100 mg·kg-1, three times per week for the first week. From the 2nd to 5th week, mice were injected intraperitoneally with TAA at 200 mg·kg-1, three times per week for the next 4 weeks. Chrysin groups were intragastrically administrated once per day to 5th week. The histopathological changes were detected by HE and Masson staining. The levels of serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were assessed by the kits. All animal experiments were approved by the Medical Ethics Committee of Affiliated Zhongshan Hospital of Dalian University (DWLL2019060). LX-2 cells were stimulated by (transforming growth factor-β, TGF-β) in vitro. The protein expressions of AMPKα, p-AMPKα, NLRP3, cysteinyl aspartate specific proteinase-1 (caspase-1), gasdermin D (GSDMD) were detected by Western blot, and the mRNA levels of collagen-Ι, α-smooth muscle actin (α-SMA), interleukin-1β (IL-1β), IL-18, caspase-1, GSDMD were analysis by reverse transcription-polymerase chain reaction (RT-PCR). Chrysin attenuated the increases in serum AST and ALT levels in the TAA group, while significantly improved the changes of liver morphology, reduced liver tissue inflammatory cell infiltration and inhibited collagens deposition. Compared with TAA group, chrysin effectively activated AMPKα phosphorylation and inhibited hepatic NLRP3 inflammasome activation. Additionally, the protein expressions and mRNA levels of IL-1β, IL-18, caspase-1 and GSDMD in chrysin groups were decreased. Chrysin inhibited the expressions of collagen-Ι and α-SMA, enhanced the phosphorylation of AMPKα, and decreased the expressions of NLRP3 and GSDMD. Therefore, chrysin may inhibit inflammatory injury and pyroptosis possibly by activating AMPK and inhibiting NLRP3 inflammasome to alleviate hepatic fibrosis.

Preparation and evaluation of quality management samples for noninvasive prenatal screening / 中华医学遗传学杂志

OBJECTIVE@#To prepare a quality control sample for non-invasive prenatal screening (NIPS) and evaluate its quality and stability.@*METHODS@#According to the biological characteristics of cell-free fetal DNA derived from the plasma of pregnant women, the simulated samples were prepared by mixing genomic DNA fragments derived from individuals with trisomy 21, trisomy 18 and trisomy 13 and background plasma. The samples were then compared with commercially made quality control products tested on various NIPS platforms and stored at -80℃, -20℃, 4℃, 24℃ and 37℃ for various periods of time.@*RESULTS@#The simulated samples have attained the expected results and could be detected on various platforms and stored at -80℃and -20℃ for at least 30 days.@*CONCLUSION@#A simulated sample was successfully prepared and possessed good stability. It can be used as the quality control sample for NIPS.

Effect of postpartum fatigue on maternal behavior in rats and its mechanism / 中华行为医学与脑科学杂志

Objective:To explore the effect of postpartum fatigue(PPF) on maternal behavior in rats and its mechanisms.Methods:Sprague-Dawley (SD) rats on the first day after delivery were randomized into the control group and the PPF group using the random number table method, with eight rats in each group.The rat model of PPF was established by forcing rats to stand in a cage with water and last for seven days.To maintain galactosis and lactation, rats and pups were caged for 90 min after every 3 h of separation.The control group was separated routinely without any stimulus.The length and body mass of the pups were recorded at birth and postnatal day 7.On the seven days after modeling, the following maternal behaviors were observed via video recordings: suckling, nesting, clicking and retrieval.The morphology of neurons in the paraventricular nucleus of hypothalamus (PVH) was observed by HE staining.The expression of oxytocin in the paraventricular hypothalamus (OxtPVH) was determined by immunofluorescence and immunohistochemistry.Western blot and qRT-PCR were performed to detect mRNA and protein expression of prolactin (PRL) in pituitary gland, respectively.Statistical analysis was performed by SPSS version 22.0, normally distributed continuous variables were compared between the two groups using an independent-sample t test, and nonnormally distributed continuous variables were compared between the two groups by Mann-Whitney U test. Results:On the seventh day after modeling, the length and weight gain of pups in the PPF group ((5.82±0.17) cm, (5.33±2.54) g)were significantly lower than those of the control group ((6.24±0.36) cm, (7.92±2.54) g, t=3.199, 2.227, both P<0.05). Compared with the control group, the rats in PPF group exhibited abnormal maternal behaviors, such as gnawing cage, biting tails, turning circles, repeatedly nesting and refusal to suckling.The results from the maternal behavioral test revealed that the latency of first pup retrieval and last pup retrieval ((39.25±3.50) s, (280.75±59.16) s) in the PPF group were significantly prolonged compared with those in the control group((19.25±7.68) s, (146.00±49.62) s, t=-4.742, -3.490, both P<0.05), the duration of nesting building ((19.50±12.69) s)and clicking ((95.50±70.55)s) in the PPF group were significantly shorter than those in the control group((68.00±37.59) s, (243.00±62.07) s; t=2.445, 3.139, both P<0.05). Compared with control group, the neurons cells of PVH in the PPF group were in disordered manner and the OxtPVH content in the PPF group decreased significantly.The mRNA (0.33(0.29, 0.38) vs 0.85(0.76, 1.76), Z=-3.576, P<0.05) and protein ((1.00±0.65) vs (4.17±0.49), t=-7.726, P<0.05) levels of PRL in PPF group were significantly decreased compared with those in the control group. Conclusion:The behaviors of holding back, nesting and licking offspring are decreased in postnatal fatigued rats.This may be related to the decreased expression of OxtPVH and PRL in hypothalamus of female rats.

Comparable prognosis in different neonatal histidine-tryptophan-ketoglutarate dosage management / 中华医学杂志(英文版)

BACKGROUND@#Histidine-tryptophan-ketoglutarate (HTK) is a solution commonly used for organ transplantation. However, there is no certified fixed regimen for on-pump heart surgery in neonates. We aimed to retrospectively evaluate the outcomes related to different HTK dosages and to analyze the safety of high-dosage perfusion.@*METHODS@#A total of 146 neonates who underwent on-pump heart surgery with single-shot HTK perfusion were divided into two groups according to HTK dosages: a standard-dose (SD) group (n = 63, 40 mL/kg 60 mL/kg). Propensity score matching (PSM) was performed to control confounding bias.@*RESULTS@#The SD group had a higher weight (3.7 ± 0.4 vs. 3.4 ± 0.4 kg, P  0.05). The incidences of post-operative complications were not significantly different between the two groups. There were no significant differences in ventilation time, intensive care unit stay, and post-operative hospital stay (P > 0.05). Follow-up echocardiography outcomes at 1 month, 3 to 6 months, and 1 year showed that left ventricular ejection fraction and end-diastolic dimension were comparable between the two groups.@*CONCLUSIONS@#In neonatal on-pump cardiac surgery patients, single-shot HD (>60 mL/kg) HTK perfusion had a comparable heart protection effect and short-term post-operative prognosis as standard dosage perfusion of 40 to 60 mL/kg. Thus, this study provides supporting evidence of the safety of HD HTK perfusion.

Swallowing Dysfunction for Old People in Nursing Homes in Ningbo / 中国康复理论与实践

Objective:To investigate the prevalence of dysphagia in old people in nursing homes in Ningbo and analyze its related factors. Methods:From August, 2017 to April, 2018, 997 people aged 60 or above were selected from eight nursing homes in four districts in Ningbo. They were investigated with general questionnaire, Kubota water swallowing test, body mass index (BMI) and Quality of Life Index (QLI), and the data were analyzed with Logistic regression. Results:There were 259 (25.98%) persons with dysphagia. Age, male, hypertension, diabetes, hyperlipidemia, coronary heart disease, stroke and fracture were risk factors for dysphagia (P < 0.01). There were 19.2% with low BMI in total, and 63.3% for dysphagia. The total score of QLI decreased with age (F = 18.706, P < 0.001), more in males than in females (t = 2.516, P = 0.012), and less in those with dysphagia than with normal swallowing (t = 7.176, P < 0.001). Conclusion:Dysphagia is prevalent in the old people in nursing homes in Ningbo, which needs early intervention.

Investigation of aflatoxins,mycobiota,and toxigenic fungi during post-harvest handling of Ziziphi Spinosae Semen / 中国中药杂志

Ziziphi Spinosae Semen is one of the Chinese herbal medicine being susceptible to aflatoxins contamination. To investigate the sources of aflatoxins contamination and toxigenic fungi species on Ziziphi Spinosae Semen,32 samples were collected from multiple steps during the post-harvest processing in this study. Aflatoxins in these samples were determined by immunoaffinity column and HPLC coupled with post-column photochemical derivatization. The dilution-plate method was applied to the fungi isolation. The isolated fungi strains were identified by morphological characterization and molecular approaches. The results showed that aflatoxins were detected in 28 samples from every step during the processing of Ziziphi Spinosae Semen. Three samples were detected with aflatoxin B_1 and 2 samples with both aflatoxin B_1 and total aflatoxin exceeding the limit of Chinese Pharmacopoeia. Especially the samples from the washing step,with the highest detected amounts of AFB_1 and AFs were reached 94. 79,121. 43 μg·kg~(-1),respectively. All 32 samples were contaminated by fungi. The fungal counts on the newly harvested samples were 2. 20 × 10~2 CFU·g~(-1). Moreover,it increased as tphreocessing progresses,and achieved 1. 16×10~6 CFU·g~(-1) after washing. A total of 321 isolates were identified to 17 genera. Aspergillus flavus was the main source of aflatoxins during the processing and storage of Ziziphi Spinosae Semen. One isolate of A. flavus was confirmed producing AFB_1 and AFB_2. The fungal count was significantly increased by composting,and Aspergillus was the predominant genus after shell breaking. The contamination level of aflatoxins was increased by composting and washing.

Chemical Constituents of Lipid-soluble Parts of Tinospora sinensis / 中国实验方剂学杂志

Objective:To isolate the chemical constituents from lipid-soluble parts of Tibetan Tinospora sinensis and identify the structure of the monomer compounds. Method:Eighty kg of dried rhizomes of T. sinensis were soaked by 80% ethanol (5 times of the dose of crude drug) for 1 hour, boiled under reflux for 1 hour,then and filtered. The above steps were repeated. The two extracts were combined,and the solvent was recovered to obtain a total extract. The obtained extract was extracted with a conventional solvent,and the solution was recovered to obtain petroleum ether extract,methylene chloride extract,ethyl acetate extract,n-butanol extract and water extract. Methylene chloride extract was isolated and purified by silicagel column chromatography,semi-preparative liquid chromatography and SephadexLH-20 chromatography; the chemical structure was identified on the basis of ESI-MS,nuclear magnetic resonance (1H-NMR and 13 C-NMR) spectroscopy data and literatures. Result:Fifteen compounds were isolated and identified respectively as n-dodecanol (1),n-hexacosanol (2),palmitic acid (3),dibutyl phthalate (4),vanillic acid (5),vanillin (6),apocynin (7),tinocordifolioside (8),medioresinol (9),isolariciresinol (10),aurantiamide (11),aurantiamide acetate (12),berberine (13),daucosterol (14),β-sitosterol (15). Conclusion:Except for 3,4,6,14,15,the other 10 compounds were isolated from the plant for the first time.

Auditory brainstem response may predict behavior threshold of children with hearing loss / 基础医学与临床

Objective To investigate the impact of degree of hearing loss on auditory brainstem response predictions of behavioral thresholds. Methods A total of 179 patients with different hearing loss degree ( a total of 311 ears) patients from January 2012 to January 2016 in the affiliated hospital of Yan’an university were treated with auditory brainstem response (ABR), performed threshold behavior test, analyed the auditory brainstem electric response and behavioral threshold correlation, the factors that affect the ABR thresholds and behavioral threshold value and threshold of ABR and correction factor as well as linear regression equation for different frequency stimulation. Results ABR threshold was positively related to threshold behavior for the different frequencies stimulation, ABR threshold may predicte threshold behavior( P＜0.05) ; ABR threshold value can predicte behavioral threshold differ-ence error and the degree of hearing loss( P＜0.05) , through the relevant correction factors can reduce the predic-tion error. Conclusions Auditory brainstem responses may effectively predicte the hearing loss in children with be-havioural thresholds, differences in degree of hearing loss influence auditory brainstem response and behavioral threshold value, constant correction coefficient is conducive to auditory brainstem electric response and facilitates accuracy of the behavior threshold.

Analysis of the impact of postpartum fatigue on the onset of lactation / 中华护理杂志

Objective To explore the impact of postpartum fatigue on the onset of lactation.Methods Totally 553 mother-infant dyads were enrolled from three hospitals in Nantong City using sampling method of probability proportional to size.The timing of the onset of lactation,delayed onset of lactation and maternal postpartum fatigue were measured during the period of hospitalization.Postpartum fatigue was measured by Fatigue Scale-14.The timing of onset of lactation was determined by matemal perception of breast fullness.One-way analysis of variance was used to compare the difference of onset of lactation in four groups with varied fatigue scores.Impact of the maternal fatigue on the onset of lactation was analyzed by multiple linear regression.Results Mothers' fatigue score was related to the onset of lactation(r-0.15,P＜0.001).Mothers with postpartum fatigue scores ranging from 9 to 14 had delayed onset of lactation compared with those with fatigue score ranging from 0 to 3 and from 4 to 5,and the differences were significant (P=0.004),and the rate of delayed onset of lactation was increased (P=0.020).Postpartum fatigue was an independent risk factor,after controlling mode of delivery,mode of anesthesia,duration of infant sucking,maternal pain,and infant sucking gesture(t=3.26,P=0.001).Conclusion Postpartum fatigue is one of reasons leading to delayed onset of lactation.Health care providers and family members should pay more attention to postpartum fatigue and take effective measures to promote a successful onset of lactation and breastfeeding.

Exploration on improvement of the compliance of colorectal cancer screening in Wujing community of Shanghai / 上海预防医学

To explore the model and method of improving the compliance of colorectal cancer screening , especially the fine sieve compliance of colonoscopy . [ Methods ] The colorectal cancer screening in Wujing community was led by Wujing Community Health Center .For the sake of impro-ving the compliance of screening , Wujing Community Health Center established a screening team and speci-fied a neighboring general hospital as screening partner .We also used the method of refining the screening steps , tracking the result in time , simplifying the procedures of colonoscopy test and making epidemiological analysis. [Results] From October 1,2012 to September 30,2013, a total of 7 507 people in Wujing community received colorectal cancer screening , of whom 427 (5.69%) were found to be positive in fecal occult blood testing (FOBT).And among them, 386 (90.40%) agreed to accept the colonoscopy test and finally 380 people (88.90%) finished the test.According to the results of colonoscopy test were detected 5 cancer patients (1.32%) and 84 precancerous lesion patients (22.11%).Further treatment rate for color-ectal cancer patients was 100.00%and for precancerous lesion patients 75.00%.The median duration was 4 days for preliminary screening positive subjects from preliminary screening to receiving positive notification and 10 days for refined screening .The median duration was 3 days for colorectal cancer patients from re-fined screening to completing treatment and 25 days for early colorectal cancer patients . [ Conclusion] The model and method adopted in Wujing community is helpful to improve the compliance of colorectal cancer screening .

Prognostic significance of coagulation disorders in children with hemophagocytic syndrome / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the prognostic significance of coagulation disorders in children with hemophagocytic syndrome (HPS).</p><p><b>METHODS</b>Thirty-five children with HPS were retrospectively studied to analyze the etiology, clinical characteristics, laboratory results and treatment outcomes.</p><p><b>RESULTS</b>After treatment, 27 of the 35 HPS patients survived, and the other 8 cases died. All cases were treated according to the HLH-2004 protocol, but etoposide (VP-16) was not used in 10 of them. The response rate in patients who received VP-16 (22/25, 88%) was significantly higher than that in those not receiving VP-16 (5/10, 50%) (P<0.05). Compared with the survival group, the dead group had significantly lower platelet count, fibrinogen level, and VP-16 utilization rate (P<0.05) but significantly longer activated partial thromboplastin time and prothrombin time (P<0.05).</p><p><b>CONCLUSIONS</b>Coagulation function can be used as an indicator of disease outcome. It is essential for improving the clinical outcome of HPS to monitor the coagulation function during treatment, detect and correct abnormalities in time, and provide treatment strictly according to the HLH-2004 protocol.</p>

Ultrastructural observation of Birbeck granules in 8 cases of Langerhans cell histiocytosis / 中华实用儿科临床杂志

Objective To study the ultrastructural features of Langerhans cell(LC),especially Birbeck granules in Langerhans cell histiocytosis(LCH) and provide some ultrastructural evidence to clarify the origin,function and stereoscopic structure of Birbeck granules.Methods Pathologic tissues of 8 LCH patients were treated with routine transmission electron microscope (TEM) procedures,fixture,dehydration,permeation,embedding,polymerization,ultramicrocut,staining and examined by TEM.Results Under the TEM,8 cases of LCH revealed typical LC cell,and LC cell in 6 cases revealed typical rod,tennnis-racquet like Birbeck granules.Some double membrane structures revealed irregular shape,sometimes accompanied with vacuole-like structure.Most of Birbeck granules locate inside the cytoplasmic membrane,in continuity with the cell membrane.There was Birbeck granule-like structure outside the cytoplasmic membrane in one case of multisystem LCH.Conclusions The ultrastructure of Birbeck granules could be various and irregular.The irregular Birbeck granule-like structure can also be useful for the diagnosis of LCH.Birbeck granules may arise from the cell membrane.The stereoscopic structure of Birbeck granules could be irregular.

Increased expressions of leptin and its receptor in the epididymis of varicocele model rats and their implications / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the expressions of leptin and its receptor in the epididymis of experimental varicocele (EV) rats.</p><p><b>METHODS</b>Forty male Sprague-Dawley rats were randomly divided into four groups: 4-week EV (n = 12), 8-week EV (n = 12), 4-week control (n = 8), and 8-week control (n = 8). EV models were established by partial ligation of the left renal vein. The expressions of leptin and its receptor in the rat epididymis were measured by immunohistochemistry, and their mRNA expressions determined by real-time quantitative PCR.</p><p><b>RESULTS</b>The expressions of leptin and its receptor in the epididymis were significantly higher in the 4- and 8-week EV groups than in the 4- and 8-week control groups (P < 0.01), with no significant difference between the two EV groups (P > 0.05). So were their mRNA expressions in the former two than in the latter two groups (P < 0.01), with no significant difference between the former two (P > 0.05).</p><p><b>CONCLUSION</b>The expressions of leptin and its receptor are markedly increased in the epididymis of varicocele rats. Leptin may be involved in the mechanisms of varicocele inducing male infertility.</p>

Clinical efficacy of desmopressin in the treatment of mild hemophilia A in children / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the effects of desmopressin (DDAVP) on coagulation factor Ⅷ (FⅧ) and activated partial thromboplastin time (APTT) in children with mild hemophilia A.</p><p><b>METHODS</b>Eighteen children with mild hemophilia A were enrolled. DDAVP (0.3 μg/kg•d) was injected intravenously for 5 days. Plasma FⅧ levels and APTT were measured before and after DDAVP treatment.</p><p><b>RESULTS</b>In 16 of 18 children with mild hemophilia A, the bleeding symptoms, including the articular or musclar hematoma, were significantly alleviated as a result of DDAVP treatment. The plasma FⅧ levels increased significantly to (27±4)% and APTT was shortened to (66±10)s 60 minutes after the first dose of DDAVP treatment. The plasma FⅧ remained at the levels of 25%-30% during 3-4 days of DDAVP treatment. Five days after DDAVP treatment, the plasma FⅧ levels decreased [(21±3)%], and APTT was prolonged when compared with 1-4 days of DDAVP treatment.</p><p><b>CONCLUSIONS</b>DDAVP treatment can increase plasma FⅧ levels and shorten APTT in children with mild hemophilia A. DDAVP is effective in the treatment of mild hemophilia A. The duration of DDAVP therapy for mild hemophilia A is recommended as 3 to 4 days.</p>

RT-PCR identification and genetic characterizations of VP1 region of human enterovirus 71 isolated in Beijing in 2008 / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the genetic characterizations of VP1 region of Human enterovirus 71 (HEV71) isolated from clinical specimens of hand, foot and mouth disease (HFMD) patients in Beijing in 2008.</p><p><b>METHODS</b>285 clinical samples were collected from HFMD patients in hospitals and day-care centers in Chaoyang district. They were performed by reverse transcription-polymerase chain reaction (RT-PCR) specific for HEV71. 10 HEV71 isolates were selected for entire VP1 coding gene amplification and sequencing.</p><p><b>RESULTS</b>129 samples were RT-PCR positive, the positive rate is 45.26%. The homology of the nucleotide and the amino acid of the 10 strains were 94.6%-99.6% and 95.9%-100%. The phylogenetic tree revealed that 10 Beijing strains clustered within the C4a evolution branch of C4 subgenotype.</p><p><b>CONCLUSIONS</b>RT-PCR played an important role in identifying HFMD outbreak in Beijing in 2008. The HEV71 strains were all belong to C4a evolution branch of C4 subgenotype with several transmission chains, and it showed that C4 subgenotype HEV71 spread in mainland China widely after 1998. The molecular epidemiology surveillance and the research of genetic characterizations of HEV71 should be strengthened in mainland China.</p>

Levels of Toll-like receptors-2,-4 on platelets in children with idiopathic thrombocytopenic purpura / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the changes and significance of Toll-like receptor-2 (TLR2) and Toll-like receptor-4 (TLR4) on platelets, CD86 on lymphocytes and concentrations of IL-2, IFN-gamma, IL-4 and IL-10 in serum in children with idiopathic thrombocytopenic purpura (ITP).</p><p><b>METHODS</b>Peripheral blood samples were collected from 24 children with acute idiopathic thrombocytopenic purpura (AITP), 21 children with chronic idiopathic thrombocytopenic purpura (CITP) and 20 normal children (control group). Expression of TLR2 and TLR4 on platelets and CD86 on lymphocytes were detected by flow cytometry. Serum concentrations of IL-2, IL-4, IL-10 and IFN-gamma were measured using ABC-ELISA.</p><p><b>RESULTS</b>The expression of CD41+TLR2+ and CD61+TLR4+ in the AITP and the CITP groups were significantly lower than those in the control group (p<0.01). The AITP group had lower expression of CD41+TLR2+ and CD61+TLR4+ than the CITP group (p<0.01). The expression of CD86+ in the AITP and the CITP groups was significantly higher than that in the control group (p<0.01). The serum concentrations of IL-2, IL-4, IL-10 and IFN-gamma in the AITP and the CITP groups were significantly higher than those in the control group (p<0.05). There was a positive correlation between CD41+TLR2+ and CD61+TLR4+ expression. CD41+TLR2+ and CD61+TLR4+ expression were negatively correlated with CD86+ expression and serum concentrations of IL-2, IL-4 and IL-10.</p><p><b>CONCLUSIONS</b>The detections of TLR2 and TLR4 on platelets, CD86 on lymphocytes and serum concentrations of IL-2, IFN-gamma, IL-4 and IL-10 are of great value in understanding the pathogenesis and predicting types of ITP in children.</p>

Effects of FMS-like tyrosine kinase 3 targeted RNA interference on proliferation and apoptosis of acute monocytic leukemia cell line THP-1 / 中华儿科杂志

<p><b>OBJECTIVE</b>FMS-like tyrosine kinase 3 (FLT3) is a receptor tyrosine kinase that is constitutively activated in (70-90)% pediatric patients with acute myeloid leukemia (AML) and appears to confer an adverse prognosis. Although several FLT3-selective small molecule inhibitors and antibodies were developed with varied degrees of success, to address the specificity and resistance, new approaches for specifically targeted FLT3 are needed and RNA interference is a promising choice. The aim of the present study was to investigate the efficacy of suppression of FLT3 induced by small hairpin interfering RNA (shRNA) on myeloproliferation and apoptosis in an acute monocytic leukemia (AMOL) cell line THP-1.</p><p><b>METHODS</b>FLT3-targeted small hairpin interfering RNA (FLT3-shRNA) was designed and synthesized by transcription system in vitro was transfected into THP-1 cells. Firstly FLT3 mRNA level was detected by semi-quantitative RT-PCR and FLT3 protein level was detected by flow cytometry (FCM) to verify the efficacy on FLT3-shRNA interference at 48 h after transfection. Cell growth viability was measured at 24 h, 48 h and 72 h after treatment with CCK-8. The distribution of cell cycle was assayed by FCM, and apoptosis was analyzed by DNA Ladder and Annexin V-FITC Staining at 48 h.</p><p><b>RESULTS</b>FLT3 targeted shRNAs was synthesized successfully and the concentration of 15 nmol/L for 48 h could obtain desirable downregulation of FLT3 expression, the inhibitory percentages of FLT3 mRNA and protein were (72.95 +/- 2.07)% and (65.39 +/- 5.57)%, respectively. The suppression of FLT3 induced by FLT3-shRNA resulted in marked inhibition of cell growth and the inhibitory percentages were (36.66 +/- 3.67)% at 48 h, (35.56 +/- 0.73)% at 72 h. FLT3-shRNA induced the inhibition of cell cycle from G(0)/G(1) phase to S phase, the percentage of sub-G(0)/G(1) phase (65.71 +/- 4.47)% was higher than those in the PBS-control group (52.23 +/- 2.98)%, NC-shRNA control group (51.81 +/- 1.44)%, P < 0.01; the percentage of S phase (25.11 +/- 2.70)% was lower than those in the PBS-control group (34.41 +/- 4.07)% and NC-shRNA control group (32.50 +/- 1.46)%, P < 0.05. Furthermore treatment with FLT3-shRNA for 48 h resulted in clear apoptosis ladder, the percentage of early apoptosis detected by Annexin V-FITC was (18.59 +/- 2.07)% which was significantly higher than that in the PBS-control group (4.00 +/- 0.50)% and the NC-shRNA control group (6.06 +/- 0.70)%, P < 0.001.</p><p><b>CONCLUSION</b>The suppression of FLT3 induced by the shRNA can effectively inhibit cell proliferation, and apoptosis induction on THP-1 cells, which indicates that this approach may bear the therapeutic potential on childhood AMOL.</p>

In vitro transcription synthesis and effects of FLT3 targeted short hairpin RNA / 中国实验血液学杂志

FMS-like tyrosine kinase 3 (FLT3) is a receptor of tyrosine kinase that is constitutively activated in most of acute myeloid leukemia patients and seems to give an adverse prognosis. In order to explore the silencing effect of FLT3 targeted short hairpin RNA (FLT3-shRNA) on acute leukaemia cell line THP-1, three FLT3-shRNAs (shRNA1, shRNA2, shRNA3) were designed and synthesized by transcription system in vitro and then transfected into THP-1 cells. FLT3 mRNA was analyzed by semi-quantitative RT-PCR, FLT3 protein was detected by Flow cytometry and immunofluorescence. The results indicated that FLT3 expression was downregulated by shRNA1 and shRNA3, and shRNA1 showed stronger inhibitory effect. At 48 hours following transfection, the inhibitory rate of 25 nmol/L shRNA1 was 72.95 +/- 2.07%, lasting 72 hours. The 5 nmol/L and more concentration of FLT3 shRNA1 could downregulate FLT3 mRNA level, which displayed a quantity-effect relation; the inhibitory rate of 15 nmol/L shRNA1 was 67.53 +/- 0.66%. FLT3 protein was located on THP-1 cell membrance, its expression was downregulated obviously by shRNA1, at 72 hours following transfection the inhibitory rate of shRNA1 was 79.67 +/- 0.66%. shRNA1 showed the best inhibitory effect on FLT3 protein, the optimal time of which was 72 hours with an inhibitory rate of 79.67%. It is concluded that FLT3-shRNA1 shows a desireable FLT3-targeted inhibitory effect, which can be used for further investigation of FLT3 mechanism or FLT3 targeting treatment.