Objective: A new type of molecular probe design method was established to improve the sensitivity and specificity of microRNA detection. Methods: This is an experimental study. The target hybridization sequence was designed on the stem side of the molecular beacon using the strand displacement principle and based on this, a new probe was designed by using the nucleic acid structure analysis software DNAman to optimize the secondary structure of the molecular probe, which was called as strand displacement molecular probe (MB-D) and MB-D plus. Taking microRNA-21 as an example, microRNA-21 and its related single nucleotide mutations were detected using conventional molecular probe (MB-C) and redesigned MBs (MB-D and MB-D plus) to analyze the differences on minimum detection limit, repeatability and specificity for microRNA detection among these three probes. Results: The minimum detection limit of MB-C for microRNA-21 was 1 nmol/L, and the minimum detection limits for MB-D and MB-D plus were 0.1 nmol/L and 0.01 nmol/L, respectively. The established MB-D plus can significantly distinguish between miR-21 and single nucleotide mutations. Conclusion The molecular probe based on the principle of strand displacement and optimized by secondary structure can significantly increase the sensitivity and specificity of the probe for microRNA detection.(Chin J Lab Med, 2018, 41: 541-546)

Objective:To explore the potential impact of lipid metabolism-related single nucleotide polymorphisms(SNP)on semen quality in men.Methods:We selected 284 semen samples from Xingtai Infertility Hospital and Hebei Human Sperm Bank collected between February and October 2023,33 from oligozoospermia(OS),97 from asthenozoospermia(AS)and 54 from oligoas-thenozoospermia(OAS)patients and the other 100 from normal men.We performed computer-assisted semen analysis(CASA)of the samples,extracted blood DNA and,using the Mass ARRAY? System,genotyped the target genes,determined the genotypes of 13 SNPs and compared their distribution,their correlation with BMI and semen quality in different groups.Results:The mutant homozygous(TT)genotype of the FADS2 rs2727270 gene seemed to be a risk factor for AS(OR=4.420,P=0.047),while the APOA2 rs5082-A allele and MC4R rs17782313 heterozygous(TC)genotype important protective factors for OS(OR=0.422 and 0.389;P=0.045 and 0.043,respectively).A significantly higher sperm concentration was found associated with the MC4R rs17782313 heter-ozygous(TC)genotype than with the homozygous(CC)genotype.Stratification analysis showed that the protective effect of the TC genotype was decreased with increased BMI and remained with the interaction of the rs5082 and rs17782313 genotypes.Conclusion:FADS2 rs2727270,APOA2 rs5082 and MC4R rs17782313 were significantly correlated with the risk of abnormal semen parameters.

Objective To analyze the chemical components of the ethanol-soluble portion of Liuwei Dihuang Decoction,and to explore its mechanism in kidney-Yin deficiency of mice.Methods Bioactive components were analyzed by HPLC,the method was used at Hypersile C1s column(4.6 mm × 250 mm,5 μm),the mobile phase consisted of methanol-water (27 ∶ 73),the detective wavelength was 236 nm,the flow rate was 1.0 mL · min-1,the column temperature was 25 ℃.The mice were divided randomly into three groups (n =10):blank group,model group,experimental group.The mice were gavaged with oral administration of corresponding medicines (9.75 g · kg-1)for nine weeks.At time 6 d after administration,in addition to that the normal group was given distilled water,the other groups were gavaged with hydrocortisone injection per day according to 50 mg · kg-1 body weight for 4 d to establish the kidney Yin deficiency mice model.The contents of cyclic adenosine monophosphate (cAMP),cyclic guanosine monophosphate (cGMP) were determined by ELISA and the follicle-stimulating hormone (FSH),estradiol (E2),testosterone (T) were determined by radioimmunoassay after administration.Results The linear range of morroniside,sweroside,paeoniflorin and loganin were 0.24-2.40 μg (r=0.999 6),0.14-1.38 μg(r =0.999 1),0.12-1.20 μg(r =0.999 1),0.24-2.40 μg(r =0.999 4) respectively.The recovery and relative standard deviation of them were (100.22 ± 1.80) ％,(100.83 ± 1.94) ％,(102.40 ± 1.47) ％,(101.40 ± 1.50) ％,and RSD were 1.80％,1.92％,1.44％,1.50％.The average content of morroniside,sweroside,paeoniflorin and loganin in Liuwei Dihuang Decoction water extract-alcohol soluble parts were calculate in terms of material were 0.20％,0.02％,0.06％,0.13％.The plasma level of cAMP in model group and experimental group were (8.20 ±0.63),(6.90 ±0.15) nmol · L-1 with significant different (P ＜0.01).The serum contents of FSH,E2,T in model group and experimental group were(0.54 ±0.10),(0.88 ±0.04)mU · mL-1;(13.93 ± 0.29),(15.48 ± 0.43) pg · mL-1;(2.23 ± 0.14),(5.63 ± 0.48) ng · mL-1 with significant different (P ＜ 0.01).Conclusion It is found that ethanol-soluble portion might be one of the bioactive component of Liuwei Dihaung Decoction to improve kidney Yin deficiency,and it mainly contains loganin,morroniside,sweroside and paeoniflorin.Its function mechanism of tonifying kidney may be related to the regulation of cAMP content and the levels of hormones in the HPG axis.

OBJECTIVE To explore the effect and mechanism of Bushen Zhuyun formula(BSZYF)on gonadotropins and gonadotropin-releasing hormone receptor(GnRHR)signaling transduction in mifepristone-induced LPD rats,and to find the scientific evidence of"heart,kidney and uterus axis"theory of TCM.METHODS SD female rats were administered mifepris-tone suspension via oral gavage,followed by pharmaceutical intervention.Then,the serum follicle-stimulating hormone (FSH)and luteinizing hormone(LH)levels were detected by ELISA assay,the FSHβand LHβexpression levels in the pitui-tary were determined by qPCR,and the GnRHR expression levels were assessed by qPCR and Western blot analysis.Simulta-neously,transcription factors,such as c-Jun,Elk-1,Egr-1,Nur77 and CREB,and transduction molecules in PKC-MAPK, Ca2+-CAM,and cAMP-PKA signaling pathways,such as PKC,p38MAPK,ERK1/2,CAM,and PKA were assessed by qPCR and Western blot analysis.RESULTS After pharmaceutical intervention,BSZYF increased the serum FSH levels sup-pressed by mifepristone(P<0.05).Moreover,the expression of c-Jun,Elk-1,Egr-1,CREB,PKA,p38MAPK,JNK,ERK and PKA were was significantly upregulated in the BSZYF group compared with that in the mifepristone group(P<0.05). However,there was no statistical difference in Nur77 and CAM expression among the blank,mifepristone and BSZYF groups (P>0.05).CONCLUSION BSZYF can regulate abnormal pituitary gonadotropin secretion and GnRHR signaling transduc-tion systems induced by mifepristone,which confirms that the prescription can regulate the whole function of the reproductive axis,and explains the consistency of"Heart,kidney and uterus axis"theory in TCM and the concept of the hypothalamus-pitu-itary-ovary axis in modern medicine

The study was aimed to investigate the pp65 antigen of human cytomegalovirus (CMV) and its clinical significance in patients revived allogeneic hematopoietic stem cell transplantation (HSCT). 104 patients received allogeneic HSCT were studied. Anticoagulant blood samples were obtained from the recipients before and after transplantation and in the convalescence. CMV pp65 antigen in leukocytes was detected by indirect immunofluorescence assay using CMV Brite Kit weekly. The results showed that among the 104 patients, 29 cases were CMV pp65 positive (27.88%). Out of 29 cases 16 were CMV antigenemia and 13 cases were CMV disease. There were 25 cases who positively responded to antiviral therapy (effective ratio 86.21%) and 4 cases died (case-fatality ratio 13.79%). The detection revealed a significant difference in the incidence of CMV infection between the patients received unrelated or haploidentical family donor HSCT (39.29%) and HLA-identical sibling donor HSCT (14.58%) (P < 0.05). The incidence rate of CMV infection in patients with 0-I grade aGVHD and patients with II-IV grade aGVHD were 19.44% and 46.88% respectively, which had significant difference (P < 0.05). There was significant difference in the occurrence of aGVHD between the patients with and without positive CMV pp65 (P < 0.05). It is concluded that infection of CMV can be detected by the CMV pp65 monoclonal fluorescence immunohistochemistry, The detection of CMV pp65 antigen in peripheral blood leukocytes as a indicator for CMV disease surveillance after HSCT, which may be used to early diagnose the CMV infection, to guide the antiviral treatment and evaluate its efficacy.
Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Child, Preschool ; China ; epidemiology ; Cytomegalovirus ; immunology ; Cytomegalovirus Infections ; diagnosis ; drug therapy ; epidemiology ; Female ; Graft vs Host Disease ; epidemiology ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Leukocytes ; virology ; Male ; Middle Aged ; Phosphoproteins ; blood ; Risk Factors ; Viral Matrix Proteins ; blood

Objective:To investigate the effect of CD8+CD28-T cells on acute graft-versus-host disease(aGVHD)after haploidentical hematopoietic stem cell transplantation(haplo-HSCT).Methods:The relationship between absolute count of CD8+CD28-T cells and aGVHD in 60 patients with malignant hematological diseases was retrospectively analyzed after haplo-HSCT,and the differences in the incidence rate of chronic graft-versus host disease(cGVHD),infection and prognosis between different CD8+CD28-T absolute cells count groups were compared.Results:aGVHD occurred in 40 of 60 patients after haplo-HSCT,with an incidence rate of 66.67％.The median occurrence time of aGVHD was 32.5(20-100)days.At 30 days after the transplantation,the absolute count of CD8+CD28-T cells of aGVHD group was significantly lower than that of non-aGVHD group(P=0.03).Thus the absolute count of CD8+CD28-T cells at 30 days after transplantation can be used to predict the occurrence of aGVHD to some extent.At 30 days after transplantation,the incidence rate of aGVHD in the low cell count group(CD8+CD28-T cells absolute count＜0.06/μl)was significantly higher than that in the high cell count group(CD8+CD28-T cells absolute count ≥0.06/μl,P=0.011).Multivariate Cox regression analysis further confirmed that the absolute count of CD8+CD28-T cells at 30 days after transplantation was an independent risk factor for aGVHD,and the risk of aGVHD in the low cell count group was 2.222 times higher than that in the high cell count group(P=0.015).The incidence of cGVHD,fungal infection,EBV infection and CMV infection were not significantly different between the two groups with different CD8+CD28-T cells absolute count.The overall survival,non-recurrent mortality and relapse rates were not significantly different between different CD8+CD28-T cells absolute count groups.Conclusion:Patients with delayed CD8+CD28-T cells reconstitution after haplo-HSCT are more likely to develop aGVHD,and the absolute count of CD8+CD28-T cells can be used to predict the incidence of aGVHD to some extent.The absolute count of CD8+CD28-T cells after haplo-HSCT was not associated with cGVHD,fungal infection,EBV infection,and CMV infection,and was also not significantly associated with the prognosis after transplantation.

AIM To investigate the effects of total Astragalus saponins on the improvement of sarcopenia in a rat model of type 2 diabetes mellitus(T2DM).METHODS The rats were divided into the normal group for a normal feeding and the model group for the feeding of high-sugar and high-fat diet combined with intraperitoneal injection of STZ to establish a T2DM model.The successful model rats were randomly divided into the model group,the metformin group(0.2 g/kg)and the total Astragalus saponins group(80 mg/kg),and given corresponding doses of drugs by gavage.After 12 weeks administration,the rats had their FBG,postprandial blood glucose(PG2h)and wet weight of skeletal muscle measured;their serum levels of INS,C-peptide(C-P),IGF-1,TNF-α and IL-1β detected by ELISA;their morphological changes of skeletal muscle observed by HE staining;their protein expressions of PI3K,p-Akt,mTOR,S6K1,FoxO1 and Murf1 in skeletal muscle detected by Western blot;and their mRNA expressions of Pi3k,Akt and mtor in skeletal muscle detected by RT-qPCR method.RESULTS Compared with the model group,the total Astragalus saponins group displayed decreased levels of FBG,PG2h,OGTT-AUC,HOMA-IR,TNF-α and IL-1β(P＜0.01);increased levels of INS,C-P,IGF-1 and wet weight of skeletal muscle(P＜0.05,P＜0.01);improved skeletal muscle atrophy and increased protein expressions of PI3K,p-Akt,mTOR and S6K1 in skeletal muscle(P＜0.05,P＜0.01);decreased protein expressions of FoxO1 and Murf1(P＜0.05,P＜0.01);and increased mRNA expressions of Pi3k,Akt and mtor(P＜0.01).CONCLUSION The improvement effects of total Astragalus saponins on sarcopenia in T2DM rats may be associated with the regulation of PI3K/Akt/mTOR and PI3K/Akt/FoxO1 pathways.

Objective To summarize the palliative care consultations proposed by the Emergency Department of Peking Union Medical College Hospital. Methods A retrospective study was conducted on 22 palliative care consultations in the Emergency Department of Peking Union Medical College Hospital from January 2017 to June 2020. Results A total of 18 patients (6 males and 12 females) received palliative care consultations in the Emergency Department,with the average age of (65±8) years (36-88 years).Specifically,10 and 6 patients received once and twice consultations,respectively,and 2 patients did not complete the consultation.Of the patients receiving palliative care consultations,15 had malignant tumors and 3 had non-neoplastic diseases.The reasons for palliative care consultations included communication (61.1%,11/18) and pain relief (61.1%,11/18).In terms of the place of death,8 patients died in the hospital and 6 patients in other medical institutions. Conclusion There is a clear demand for palliative care consultation in the Emergency Department of Peking Union Medical College Hospital,and the consultation can bring help to both emergency doctors and patients.
Male ; Female ; Humans ; Middle Aged ; Aged ; Palliative Care/methods* ; Retrospective Studies ; Referral and Consultation ; Hospitals ; Emergency Service, Hospital

The samples of Huangqi injection (HI) were analyzed by liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (LC-TOF-MS), and both positive and negative ion modes were employed to obtain the LC-TOF-MS analysis information of chemical compounds in HI. Then the mass defect filtering (MDF) approach, which was developed based on the previously published articles, was utilized to rapidly screen the astragalosides from the obtained LC-TOF-MS data. Each screened astragaloside was confirmed by the presence of no less than 2 quasi-molecular ions. All the screened astragalosides were then tentatively assigned according to the parent ion and daughter ion information. Finally, a total of 62 astragalosides were screened and characterized from the HI samples, including 15 new detected ones. The identification results indicated that acetylation, hydrogenation, dehydrogenation, methoxylation and hydration might be the major conversion reactions involved in the formation of the astragalosides. The LC-TOF-MS-based MDF approach was proved to be a feasible and efficient tool to screen the chemical constituents in complex matrices such as herbal medicines.

End-stage patients experience unbearable pain because of refractory symptoms.Palliative sedation is a form of palliative care which relieves patients' agony by lowering their consciousness.Standard palliative sedation can help patients die with dignity.It is distinct from euthanasia and does not alter the survival of patients.Sufficient palliative care is the premise of palliative sedation.Repeated and detailed clinical evaluation,as well as multidisciplinary involvement,is necessary for the standardized implementation of palliative sedation.Here,we proposed the standard process and specifications of palliative sedation in Peking Union Medical College Hospital.Furthermore,we reported a case of palliative sedation for an advanced cancer patient with refractory delirium and living pain to demonstrate its application in clinical practice.
Humans ; Anesthesia ; Pain ; Hospitals ; Palliative Care ; Universities