Objective To explore the optimal storage methods to maintain serum clozapine (CZP) and N-desmethylclozapine (N-CZP) concentration in a long-term stablility. Methods Ten fresh clinical blood samples were collected, and each sample's serum was separated at volume of 0.5 ml into five pieces of 10 ml conic glass cen-trifuge tubes with cocks; then CZP and N-CZP were beth assayed with HPLC in samples collected on sampling day, stored at 4 ℃ and -20 ℃ for 12 weeks and 24 weeks,respectively. Results There was no significant difference in CZP among samples stored at 4 ℃ for 12 weeks[(525.1±124.3) μg/L] ,at -20 ℃ for 12 weeks[ (535.5± 126.7) and 24 weeks[ (532.5±126.7) μg/L] ,as well as collected on sampling day[(542.7±135.0) μg/L] (P>0.05); There was also no significant difference in N-CZP among samples stored at -20 ℃ for 12 weeks [(226.2±50. 7) μg/L] and 24 weeks[ (224.9±44.8) μg/L] and collected on sampling day[(236.9± 66.6) μL] (P>0.05). Conclusion CZP and N-CZP concentration would be maintained stable within 24 weeks under the refrigerated storage temperature of -20 ℃ at a fixed volume.

OBJECTIVE:To investigate the healing-promoting effect of total tanshinone on sore and ulcer of model rats. METH-ODS:Staphylococcus aureus levitation liquid was made to reproduce model rats with sore and ulcer,sc. 50 rats were randomly di-vided into normal group(equal volume of CMC-Na solution),model group(equal volume of CMC-Na solution),ampicillin group (positive drug 20 mg/kg)and total tanshinone high and low dose groups(200 and 100 mg/kg),ig,once a day for continuous 12 d. The general conditions were observed. The content of lysozyme(LZM)and immunoglobulins G(IgG)in serum were tested af-ter 12 d;the content of LZM and IgG in purulent secretion of model rats were respectively tested after dosing of 4 d,8 d and 12 d. The integral of wound deterioration were evaluated. RESULTS:Compared with normal control group,the index was worse in model group. Compared with model group,the appetite and activity amount of total tanshinone in high dose group (200 and 100 mg/kg)were increased,with no secretion on the wound and a clean escharosis;content of LZM and IgG in serum of rats in in to-tal tanshinone high and low dose group were increased after 12 d;also,the content of LZM and IgG in purulent secretion of rats in total tanshinone high and low dose groups were increased after dosing of 4 d,8 d,and 12 d;the integral of wound deterioration in total tanshinone high dose group were decreased after dosing of 8 d and 12 d,with significant difference (P<0.01 or P<0.05). CONCLUSIONS:Total tanshinone can promote the wound healing on sore and ulcer of model rats by a mechanism that is related to the content of LZM and IgG in serum and purulent secretion.

Objective To evaluate the therapeutic effect of thymosin alpha -1 with glucocorticoid in treat-ment of HBV -related hepatic failure.Methods 130 cases were randomly divided into two groups,they were all giv-en antiviral therapy,protect liver,anti -inflammatory,yellow suit the back support,etc.comprehensive treatment;and patients in treatment group were given thymosin alpha -1 with methyl -prednisone intravenously at the early stage of treating process,and then observed the clinic situation and cure rate of those sufferers,The biochemiccal indicator, PTA and blood serum HBV DNA capacity ending with the period of 4 weeks were tested.Results In both groups,the TBil,TC in serum had apparently improved compared with the baseline after the medication,the difference was signifi-cant (t =3.12,P <0.05 and t =3.05,P <0.05).The time of gastrointestinal symptoms improvement and bilirubin subsided time in treatment group were significantly shorter than those of the control group(t =3.34,P <0.01 and t =4.52,P <0.01 ).During the treatment,there was no significant adverse reaction,and there were no differences between two groups in Alt,PTA,HBV DNA,infection,gastrointestinal bleeding,hepatic encephalopathy and hepatore-nal syndrome.The effective rate of treatment group was 75.2%,which was higher than 50.3% of the control group (χ2 =11.02,P <0.01).Conclusion Patients with HBV -related hepatic failure of short -term application of thy-mosin alpha -1 with glucocorticoid treatment,can quickly improve symptoms,greatly improve the efficiency of survival rate,shortem hospitalization period,reduce side effects and enhance security.

Objective To explore the dynamic changes of serum S100B and glial fibrillary acidic protein (GFAP) levels and their clinical significance in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).Methods By means of enzyme-linked immunno-sorbent assay (ELISA),the serum S100B and GFAP levels from 33 DEACMP patients were assayed,and the condition changes were analyzed with three types of scale:the activity of daily living scale ( ADL),information-memory-concentration test (IMCT) and Hasegawa' s dementia scale(HDS).The comparison with 32 patients of acute carbon monoxide poisoning without DEACMP was also conducted.Results (1) The serum S100B( (0.60 ±0.21)ng/ml) and GFAP( (226.58 ±90.05 )ng/ml) in DEACMP group at acute stage were significantly higher than those in acute-CO-poisoning group ( (0.50 ± 0.20) ng/ml,( 183.04 ± 73.01 ) ng/ml) and those in DEACMP group at convalescent stage ( (0.51 ±0.16) ng/ml,(183.25 ±81.76)ng/ml) (all P values ＜0.05).(2)In DEACMP group,the serum S100B and GFAP at acute and convalescent stages were significantly correlated (at acute stage:r=0.466,P=0.006; at convalescent stage:r=0.365,P=0.037 ).(3)The S100B and GFAP in ineffective DEACMP patients at acute stage were significantly higher than those in the other groups ( all P values ＜ 0.05 ).(4) In DEACMP group,the ADL,HDS and IMCT scores( (45.21 ± 9.69),(8.26 ± 6.31 ),(9.91 ± 7.52) ) at acute stage were significantly different from those at convalescent stages( (33.67 ± 13.62),( 15.91 ± 10.83),( 19.06 ± 10.37 ) ) ( all P values ＜0.01 ).Conclusion There is secondary brain insult (SBI) in DEACMP; glial activation may play an important role.The S100B and GFAP levels may be associated with the prognosis of DEACMP.

Objective To explore the effect of Activating Blood to Resolve Stagnation on the expression of CD34 and vascular endothelial growth factor (VEGF) in rats with acute myocardial infarction. Methods 32 Sprague-Dawley rats were randomly divided into sham operation group (A, n=8), model group (B, n=8), Xuesaitong Injection + granulocyte colony- stimulating factor (G- CSF) group (C, n=8) and G-CSF group (D, n=8). Corresponding medicine was given to each group 3 hours after modeling, for 6 days. Pathomorphological changes were observed through HE staining, and the expression of CD34, VEGF and Ki-67 were observed through immunohistochemical staining. Results The expressions of CD34, VEGF and Ki-67 were higher in groups B, C and D than in group A (P<0.05), and were higher in group groups C and D than in group B (P<0.05). The expressions of CD34 and VEGF were higher in group C than in group D (P<0.05). However, there was no significant difference in the expression of Ki-67 between 2 groups (P>0.05). Conclusion The expression of CD34 and VEGF increases with Activating Blood to Resolve Stagnation method, which is superior to using G-CSF only. Activating Blood to Resolve Stagnation may play an important role in the treatment of acute myocardial infarction.

OBJECTIVE: To assess the value of copy number variation sequencing (CNV-seq) and karyotyping in the prenatal diagnosis for carriers of balanced translocations. METHODS: Clinical records of 135 amniocentesis samples of balanced translocation carriers undergoing simultaneous CNV-seq and karyotyping were analyzed. Chromosomal aberrations were defined as those can definitely lead to birth defects definitely, which included chromosomal numerical abnormality, large deletion/duplication and pathogenic copy number variations (pCNVs). RESULTS: The detection rates for karyotyping and CNV-seq were 4.44% (6/135) and 5.93% (8/135) respectively, and the latter had a detection rate of 1.48(2/135) higher than the former. A total of 68 fetal chromosomal translocations were detected by karyotying analysis. CONCLUSION For couples carrying a balanced translocation, simultaneous CNV-seq and karyotyping is conducive to the detection of fetal chromosomal abnormalities and genetic counseling.
Chromosome Aberrations ; Chromosome Disorders/genetics* ; DNA Copy Number Variations ; Female ; Humans ; Karyotyping ; Pregnancy ; Prenatal Diagnosis ; Translocation, Genetic

Objective:To analyze the genetic etiology of 487 fetuses with increased nuchal translucency (NT) using copy number variant sequencing (CNV-seq) and explore the relationship between increased NT and chromosomal abnormality.Methods:A retrospective study was performed on 487 fetuses with increased NT who received CNV-seq in the First Affiliated Hospital of Zhengzhou University from January 2018 to December 2020. These fetuses either had NT of ≥3.0-<3.5 mm (Group A, n=129) or ≥3.5 mm (Group B, n=358), the distribution and incidence of chromosomal abnormalities in the two sets of fetuses were analyzed using Chi square test or Fisher's exact test. Results:Fetuses with abnormal chromosomes accounted for 25.9%(126/487) of cases, including 107 with chromosome aneuploidy (22.0%) and 19 with pathogenic or likely pathogenic copy number variation (CNV, 3.9%). The detection rate of fetal aneuploidy in Group B was higher than that in Group A [14.0% (18/129) vs 24.9% (89/358), χ2=6.58, P=0.010]. However, no significant difference was observed regarding the detection rate of pathogenic or likely pathogenic CNV between the two groups ( χ2=0.30, P=0.584). Conclusions:The risk of fetal chromosome aneuploidy increased with NT thickness, but not with pathogenic or likely pathogenic CNV, which needed further verification due to the small sample size. CNV-seq is an option to detect the conventional detection methods for the genetic etiology of NT thickening fetuses.

Objective:To investigate the effect of Asarinin on the survival time of transplanted heart after allogeneic heterotopic heart transplantation and to further verify the anti-immune rejection effect of Asarinin in spleen and peripheral blood.Methods:Using 64 Wistar rats as donors, 64 SD rats as recipients to establish the allogeneic heterotopic heart transplantation model in rats.After successful transplantation, 64 rats were use simple randomization divided into control group, cyclosporine A(CsA) group, Asarinin group and half CsA + half Asarinin group with 16 rats in each group.CsA group was given 5 mg/kg by gavage; Asarinin group was given 25 mg/kg; half dose group was given CsA 2.5 mg/kg+ Asarinin 12.5 mg/kg and the control group was given the same volume of normal saline by gavage.After administration for 1 week, half of them were used to observe the survival time.The other half of the rats were fully anesthetized with chloral hydrate, spleen and peripheral blood were taken.Half of the spleen was taken to observe the slices under the microscope.The other half of spleen was used RT-PCR to detect the relative expression of IFN-γ and IL-4.The expression of co-stimulatory molecules CD80, CD86 and CD40 in peripheral blood were detected by flow cytometry.Results:Survival time of transplanted heart was control group (8.4±0.9), CsA group (30.5±8.3), Asarinin group (16.5±4.3) and half-dose group (26.1±5.2) days.Compared with control group, survival time of heart transplantation became prolonged in all groups and the difference was statistically significant ( P<0.05). HE staining of splenic tissue showed that, as compared with control group, the injury of each group was alleviated.The relative expression of IFN-γ in spleen was control group (1.055±0.083), CsA group (0.396±0.038), Asarinin group (0.833±0.094) and half-dose group (0.862±0.104). The last three groups were lower than control group and the difference was statistically significant ( P<0.05). The relative expression of IL-4 in spleen was control group (1.429±0.234), CsA group (3.808±0.729), Asarinin group (2.209±0.306) and half-dose group (2.323±0.321). The last three groups all spiked as compared with control group and the difference was statistically significant ( P<0.05). The expressions of CD80, CD86 and CD40 in peripheral blood were control group (98.21±0.54), (85.78±0.89) and (96.36±0.66), CsA group (89.26±0.36), (56.86±2.32) and (88.11±1.61), Asarinin group (94.19±0.47), (79.01±1.12) and (87.86±1.67) and half-dose group (94.87±0.74), (80.81±0.98) and (89.71±0.97) respectively.The last three groups were lower than control group and the difference was statistically significant ( P<0.05). Conclusions:Asarinin can prolong the survival time of transplanted heart after allogeneic heterotopic heart transplantation in rats, inhibit the immune injury of spleen after allogeneic heterotopic heart transplantation in rats, decrease IFN-γ in spleen, increase IL-4 in spleen and inhibit the expression of peripheral blood costimulatory molecules CD80, CD86 and CD40.

As a natural plant source of artemisinin,a first-line drug against malaria,Artemisia annua directly affects the extraction process of artemisinin and the source of artemisinin. At present,traditional breeding methods combined with tissue culture are often used to breed high-yield artemisinin-containing new varieties of A. annua. However,the breeding method has the disadvantages of low efficiency and continuous selection. In this study,heavy ion beam irradiation technology was used to observe the specific germplasm resources of A. annua,and the morphological characteristics,agronomic traits and artemisinin content were used as indicators to observe the selection materials and materials. The cultivated new varieties were compared with trials and regional trials. In addition,the new variety of A. annua was identified by SRAP molecular marker technology. The results showed that the new variety of A. annua, " Kehao No.1",had an average yield of 235. 0 kg of dry leaf per mu,which was more than 20% higher than that of the control. Especially,the average artemisinin content was 2. 0%,which was 45% higher than that of the control,and the " Kehao No.1" has high anti-white powder disease,high-yield and high-quality new varieties. Therefore,mutagenic breeding of heavy ion beam irradiation can significantly improve the yield and artemisinin content of the " Kehao No. 1" and it has a good promotion value.
Artemisia annua/genetics* ; Artemisinins/analysis* ; Heavy Ions ; Mutagenesis ; Phenotype ; Plant Breeding ; Plants, Medicinal/genetics*

In recent years， there have been both achievements and criticisms in using the methods of evidence-based medicine to evaluate the efficacy of traditional Chinese medicine （TCM）， which is mainly due to the differences between TCM and Western medicine. To facilitate the clinical evidence-based evaluation in TCM， this paper analyzes the challenges faced in TCM clinical evaluation， particularly in the diagnosis， clinical intervention， and efficacy assessment methods. Considering the current state of TCM clinical evaluation and our clinical research experience， we believe that establishing and refining the TCM disease-syndrome diagnostic system is a prerequisite for the practice of clinical evidence-based evaluation in TCM. Furthermore， this paper discusses the specific connotation， development， and challenges of the disease-syndrome diagnostic system， especially the choice of TCM disease name or modern medical disease name in this system. Then， the clinical application scenarios are expounded from ''TCM disease name + syndrome differentiation'' and ''Western medicine disease name + syndrome differentiation''. Moreover， this paper proposed solutions for practical issues such as the standardization of disease and syndrome diagnosis， selection of clinical evaluation methods， and application of evidence-based approaches in clinical evaluation. Establishing the criteria for the disease-syndrome diagnostic system is crucial for the determination of clinical intervention regimen， the selection of clinical research methods， and the establishment of evaluation indicators， which are essential for generating high-quality clinical evidence. To sum up， this paper reviews the development and current situation of the disease-syndrome diagnostic system and proposes an exploratory approach for the standardization and application of this system in clinical evidence-based evaluation. This approach aims to facilitate the integration of TCM with modern clinical practice， thereby achieving standardized evaluation of TCM efficacy and deepening the integration of TCM with evidence-based medicine.