Object To develop a best approach for the rapid propagation of Lilium brownii F.E. Brown var. viridulum Baker by tissue culture. Methods Different parts of the bulb were tried as the explant and cultured on different culture media with the additon of different portions of various hormones at various cultural conditions. Results The best medium for the culture of explant bulb was MS+NAA 0.5 mg/L and GA 3 2.0 mg/L and the highest induction rate was at the lowest part of the scale leaves, which attained 92.5%. The small bud can further differentiate to form secondary small buds. By liquid-quivering culturing, the weight increase can be accelerated. Conclusion Tissue culture of L. brownii var. viridulum can achieve its rapid propagation, resulting in the possibility for its industrial production.

OBJECTIVETo observe the effect of the extracts from rabbit skin inflamed by vaccinia virus for injection, a commercially available agent (Analgecine), in the treatment of postherpetic neuralgia (PHN) in aged patients and its influence on serum interleukin-6 (IL-6) level.

METHODSA total of 44 elderly patients with PHN were randomly divided into two groups. In addition to the conventional antiviral therapy, patients in group A were given analgecine, and those in group B given indomethacin tablet. After the completion of the 14-day treatment course, the therapeutic effects were evaluated using visual analogue scale (VAS), and the changes in serum IL-6 levels after treatment were measured.

RESULTSThe score of VAS decreased significantly after treatment in group A (P<0.01), but not in group B (P>0.05). In both groups, the serum IL-6 level before treatment was significantly higher than that in normal control subjects (P<0.001), and lowered significantly after treatment (P<0.01) in group A, but not in group B (P>0.05).

CONCLUSIONThe extracts from rabbit skin inflamed by vaccinia virus have definite effects for rapid pain relief, and can effectively decrease the serum IL-6 level in elderly patients with PHN.

Aged ; Aged, 80 and over ; Analgesics ; therapeutic use ; Animals ; Female ; Humans ; Inflammation ; virology ; Interleukin-6 ; blood ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Neuralgia, Postherpetic ; drug therapy ; Pain Measurement ; Rabbits ; virology ; Skin ; virology ; Vaccinia virus

OBJECTIVE To explore the effects of the expression of serum Sirtuin-1 （SIRT1） on therapeutic efficacy of sodium valproate （VPA） in the treatment of epilepsy patients. METHODS Fifty-four epileptic patients were collected from the Affiliated Baiyun Hospital of Guizhou Medical University from Mar. to Oct. 2021 as the research objects， and fifty healthy people were also collected during corresponding period as baseline reference samples. The patients whose relative mRNA expression of SIRT1 was lower than the baseline were selected as SIRT1 low-expression treatment group， and the patients whose that expression was higher than the baseline as SIRT1 high-expression treatment group.All patients were treated with low dose （12 mg/kg or about 600 mg/day） of VPA for 3 months， and the clinical efficacy was evaluated. The dosage of VPA in patients with ineffective gzwkj2021-468） epilepsy control should be increased to 15 mg/kg or about 800 mg/day for another 3 months as SIRT1 high-expression intensive treatment group and SIRT1 low-expression intensive treatment group. Clinical efficacies were evaluated. The blood concentration and liver function indexes of the patients were detected after 3 months of treatment and 3 months of intensive treatment. RESULTS Before treatment， among 54 epileptic patients， 31 epileptic patients had low expression of SIRT1， and 23 had high expression of SIRT1. After 3 months of low-dose VPA treatment， within effective blood concentration of VPA， effective control rate of patients in SIRT1 high-expression treatment group was significantly lower than SIRT1 low-expression treatment group （P＜0.05）. After 3 months of intensive treatment， the effective control rate of patients SIRT1 high-expression intensive treatment group was significantly higher than SIRT1 high expression treatment group （P＜0.05）. No abnormality was found in liver function indexes during VPA treatment. CONCLUSIONS Epilepsy in patients with high expression of serum SIRT1 may be more difficult to control when VPA is within the effective blood concentration range； when the VPA dose is effectively increased， the effective control rate of epilepsy can be improved.

OBJECTIVE: To construct cytarabine-resistant acute myeloid leukemia (AML) cell lines, and explore the correlation between Sirt1, PGC-1α expression levels and drug resistance. METHODS: Human acute promyelocytic leukemia Kasumi-1 cells were induced by the method of gradually increasing the concentration of Ara-C drug. The IC₅₀ value of Kasumi-1 cells before and after drug addition was detected by CCK-8 method, so as to construct Ara-C resistant cell lines. The expression levels of Sirt1 and PGC-1α mRNA in Kasumi-1 drug-resistant cell lines and their parental cell lines were detected by real-time fluorescence quantitative PCR, and the expression levels of Sirt1 and PGC-1α protein in kasumi-1 drug-resistant cell lines and their parental cell lines were detected by Western blot. RESULTS: The constructed Kasumi-1 cell line had common morphological characteristics of drug-resistant cell lines under microscope, and the drug resistance index was greater than 5, indicating that Kasumi-1 drug-resistant cells had good drug resistance after the construction. The RT-qPCR and Western blot assays showed that the expression levels of Sirt1 and PGC-1α mRNA and protein in the drug-resistant cell lines were higher than those of the parental cell lines (P<0.001). CONCLUSION AML cell lines resistant to Ara-C can be successfully induced by the method of gradually increasing the concentration, and the co-high expression of Sirt1 and PGC-1α may mediate the drug resistance of AML cells.
Cell Line ; Cytarabine/pharmacology* ; Drug Resistance ; Humans ; Leukemia, Myeloid, Acute/genetics* ; RNA, Messenger/genetics* ; Sirtuin 1

Mycoplasma pneumoniae is one of the common pathogens of community-acquired pneumonia, which not only can cause respiratory performance, but also can cause extrapulmonary performance.Among them, arteriovenous embolism is difficult to identify early, lacking specific clinical manifestations.Once embolism occurs in the heart, lung, brain, etc, the condition is critical and can cause death or disability in a short time, which greatly threatens children′s health.Thrombosis is often accompanied by an increase in D-dimer, but the level of D-dimer fluctuates greatly.Therefore, it is still difficult to identify and diagnose the hypercoagulable state early in the clinic and evaluate the risk factor of thrombosis.

OBJECTIVE: To explore the genetic basis for a Chinese pedigree with suspected mitochondrial functional defects through combined next-generation sequencing (NGS), copy number variation sequencing (CNV-seq), and mitochondrial DNA (mtDNA) sequencing. METHODS: Clinical data of the proband and his family members were collected. The patient and his parents were subjected to family-trio whole-exome sequencing (WES), CNV-seq and mtDNA variant detection. Candidate variant was verified by Sanger sequencing. RESULTS: Trio-WES revealed that the proband has carried compound heterozygous variants of the NDUFS1 gene, including a paternally derived c.64C>T (p.R22X) nonsense variant and a maternally derived c.845A>G (p.N282S) missense variant. Both variants may cause loss of protein function. No variant that may cause the phenotype was identified by CNV-seq and mtDNA variant analysis. CONCLUSION Children with suspected mitochondrial disorders may have no specific syndromes or laboratory findings. A comprehensive strategy including mtDNA testing may facilitate the diagnosis and early clinical interventions.
Child ; China ; DNA Copy Number Variations ; Electron Transport ; Humans ; Mutation ; NADH Dehydrogenase/genetics* ; Pedigree

OBJECTIVE: To analyze the clinical phenotype and genetic variant of a child with Snijders Blok-Campeau syndrome (SBCS). METHODS: A child who was diagnosed with SBCS in June 2017 at Henan Children's Hospital was selected as the study subject. Clinical data of the child was collected. Peripheral blood samples of the child and his parents were collected and the extraction of genomic DNA, which was subjected to trio-whole exome sequencing (trio-WES) and genome copy number variation (CNV) analysis. Candidate variant was verified by Sanger sequencing of his pedigree members. RESULTS: The main clinical manifestations of the child have included language delay, intellectual impairment and motor development delay, which were accompanied with facial dysmorphisms (broad forehead, inverted triangular face, sparse eyebrows, widely spaced eyes, narrow palpebral fissures, broad nose bridge, midface hypoplasia, thin upper lip, pointed jaw, low-set ears and posteriorly rotated ears). Trio-WES and Sanger sequencing revealed that the child has harbored a heterozygous splicing variant of the CHD3 gene, namely c.4073-2A>G, for which both of his parents were of wild-type. No pathogenic variant was identified by CNV testing. CONCLUSION The c.4073-2A>G splicing variant of the CHD3 gene probably underlay the SBCS in this patient.
DNA Copy Number Variations ; Heterozygote ; Pedigree ; Phenotype ; RNA Splicing ; Mutation

OBJECTIVES: The incidence of infertility is increasing, more than 30% of them having related abnormal tubal patency. Four-dimensional (4D) hysterosalpingo-contrast sonography (HyCoSy) overcomes the shortcomings of 3D HyCoSy in the diagnosis of tubal patency, showing high specificity and accuracy. In addition, 4D HyCoSy discards iodine allergy and X-ray radiation and possesses easy-operating, contributing to good acceptance in clinical practice. However, there is no research to explore the imaging standards related to the possibility of natural pregnancy after 4D HyCoSy. If a predictive model of postoperative natural pregnancy was established using the analysis of clinical data combined with imaging characteristics of 4D HyCoSy of patients with tubal factor infertility, clinical decision-making can be wisely guided in the future. This study aims to establish a predictive model of natural pregnancy after 4D HyCoSy based on clinical data and imaging characteristics of patients with tubal factor in fertility. METHODS: A retrospective study was conducted for patients who were diagnosed with tubal factor infertility in Hunan Guangxiu Hospital from February 2017 to May 2018. The patients ought to possess complete 4D HyCoSy imaging data and at least one-side-unobstructed fallopian tube. General clinical data and imaging data were collected. Pregnancy outcome was followed up till 3 months after 4D HyCoSy. According to pregnancy outcome, patients were divided into a pregnancy group and a non-pregnancy group. Binary logistic regression was used to analyze the correlation between various variables and natural pregnancy after 4D HyCoSy. The variables with significant difference (P<0.05) in single-factor logistic regression were included in the natural pregnancy probability prediction model. The classification accuracy was further verified with 10-fold cross-validation. RESULTS: A total of 1 085 patients with clinically suspected tubal factor infertility who met the requirements and followed the doctors' prescription were collected. Clinical characteristics (age and duration of infertility) and 4D HyCoSy imaging characteristics (thickness of endometrium from the 3rd to the 7th day after the end of menstruation, visualization of the left fallopian tube, the diffusion of contrast agent around the left ovary, and the diffusion of contrast agent around the right ovary) were independent predictors for natural pregnancy 3 months after 4D HyCoSy. A natural pregnancy probability prediction model was established with the area under the curve (AUC) verified by the 10-fold cross-validation all greater than 0.75, and the best AUC was 0.868. The Q value obtained by the prediction model was the probability of natural pregnancy, and the cutoff value was 0.5. When the Q value was greater than 0.5, it was recommended to attempt natural pregnancy for 3 months, and when the Q value was less than 0.5, in vitro fertilization was adviced. CONCLUSIONS A predictive model for the evaluating probability of natural pregnancy in women with tubal factor infertility after 4D HyCoSy is successfully established based on the analysis for clinical data and imaging characteristics. This model shows a great potential in assisting clinical decision making.
Humans ; Female ; Pregnancy ; Contrast Media ; Retrospective Studies

OBJECTIVE: To determine and verify the correlation formula of age estimation using the content of signal joint T-cell receptor excision DNA circle (sjTREC) in human peripheral blood and to discuss its application value in forensic biological practice. METHODS: The samples of peripheral blood stains were collected from 30 healthy unrelated individuals whose ages were known. The DNAs were extracted from the samples stored at room temperature after 4 weeks. The content of sjTREC was measured by real-time fluorescent quantitative PCR technique, and the TATA box binding protein (TBP) was selected as reference genes. The age of each sample was predicted with the formula which was Age = -7.181 5 Y-42.458 +/- 9.42 (Y = dCtTBP-sjTREC), and the result was compared with the real age of each individual to determine the accuracy of the formula. RESULTS: sjTREC and TBP gene were detectable in all 30 samples of peripheral blood. The contents of sjTREC in human peripheral blood showed a decreasing tendency with aging. The accuracy rate for the age estimation by this method was 76.67%. CONCLUSION The method for the age estimation with the content of sjTREC was simple, fast, sensitive, and good species specific with important potential application prospect.
Adolescent ; Adult ; Aged ; Aging/blood* ; Blood Stains ; Child ; Child, Preschool ; DNA/genetics* ; DNA Primers/genetics* ; Female ; Forensic Genetics/methods* ; Gene Rearrangement, T-Lymphocyte/genetics* ; Humans ; Infant ; Male ; Middle Aged ; Real-Time Polymerase Chain Reaction/methods* ; Sensitivity and Specificity ; TATA-Box Binding Protein/genetics* ; Young Adult

This study aimed to provide scientific evidence for predicting quality markers(Q-markers) of Elephantopus scaber by establishing UPLC fingerprint of E. scaber from different geographical origins and determining the content of 13 major components, as well as conducting in vitro anti-cancer activity investigation of the main components. The chromatographic column used was Waters CORTECS UPLC C_(18)(2.1 mm×150 mm, 1.6 μm), and the mobile phase consisted of acetonitrile and 0.1% formic acid solution(gradient elution). The column temperature was set at 30 ℃, and the flow rate was 0.2 mL·min~(-1). The injection volume was 1 μL, and the detection wavelength was 240 nm. The UPLC fingerprint of E. scaber was fitted using the Similarity Evaluation System for Chromatographic Fingerprint of Traditional Chinese Medicine(2012 edition) to determine common peaks, evaluate similarity, identify and determine the content of major components. The CCK-8 assay was used to explore the inhibitory effect of the main components on the proliferation of lung cancer cells. The results showed that in the established UPLC fingerprint of E. scaber, 35 common peaks were identified. Thirteen major components, including neochlorogenic acid(peak 1), chlorogenic acid(peak 2), cryptochlorogenic acid(peak 3), caffeic acid(peak 4), schaftoside(peak 6), galuteolin(peak 9), isochlorogenic acid B(peak 10), isochlorogenic acid A(peak 12), isochlorogenic acid C(peak 18), deoxyelephantopin(peak 28), isodeoxyelephantopin(peak 29), isoscabertopin(peak 31), and scabertopin(peak 32) were identified and quantified, and a quantitative analysis method was established. The results of the in vitro anti-cancer activity study showed that deoxyelephantopin, isodeoxyelephantopin, isoscabertopin, and scabertopin in E. scaber exhibited inhibition rates of lung cancer cell proliferation exceeding 80% at a concentration of 10 μmol·L~(-1), higher than the positive drug paclitaxel. These results indicate that the fingerprint of E. scaber is highly characteristic, and the quantitative analysis method is accurate and stable, providing references for the research on quality standards of E. scaber. Four sesquiterpene lactones in E. scaber show significant anti-cancer activity and can serve as Q-markers for E. scaber.
Humans ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal/chemistry* ; Asteraceae/chemistry* ; Lung Neoplasms/drug therapy*