Based on the literature study,the thoughts and possible therapeutic mechanism in treating male infertility from the perspective of spleen and kidney by regulating intestinal flora were explored.Disturbance of intestinal flora is one of the important factors leading to the development of male infertility,and the spleen and kidney have certain similarities to intestinal flora in the physiological function and pathological changes.Moreover,tonifying the kidney and strengthening the spleen can regulate the intestinal flora by fostering the growth of beneficial bacteria,inhibiting the reproduction of pathogenic bacteria,and protecting the barrier of the intestinal mucosa.Therefore,the possible therapeutic mechanisms in treating male infertility with the prescriptions for tonifying the kidney and strengthening the spleen to regulate intestinal flora are as follows:inhibiting the expression of inflammatory factors to reduce the inflammatory reaction of testicular tissues;improving the antioxidant capacity to alleviate the damage of spermatozoa caused by oxidative stress,and improving the bad mood to alleviate the impact of psychological stress on the reproductive system.The exploration of the thoughts for treating male infertility from the perspective of spleen and kidney by regulating intestinal flora may provide a new entry point for modern Chinese medicine clinical treatment of male infertility.

Objective: To examine the developmental trajectory of fine motor ability in schoolage children with attention deficit hyperactivity disorder (ADHD) for two years, so as to provide scientific evidence to promote motor development in ADHD children. Methods: From April to June 2019, 31 children aged 6-8 years old were selected from a public elementary school. They were diagnosed with ADHD by two psychiatric professionals according to the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-V) criteria. Additionally, 31 typical developmental children, matched for age, sex and IQ with the ADHD group, were recruited as the control group. Fine motor ability was assessed with tasks of hand manual dexterity in Movement Assessment Battery for Children-2 (MACB-2), and a followup assessment was conducted from April to June 2021. The development changes of fine motor ability between two groups of children were compared by using t test and repeated measures analysis of variance. Results: Between baseline and followup periods after two years, the total score of hand fine motor in the ADHD group did not show significant improvement (7.4±3.0, 8.0±3.4; t=-1.05, P>0.05), while there was a small effect size improvement in typically developing control group (9.5±2.1, 10.5±2.4; t=-2.12, effect size=0.38, P<0.05). Followup after two years, coin/peg throwing scores with dominant hand improved between ADHD group and control group (7.0±3.3, 9.5±3.2; 8.4±2.8, 11.6±1.6) (t=-3.74, -6.33, P<0.01; effect size=0.67, 1.14), with a smaller improvement in the ADHD group. The score for threading beads/threads decreased in between ADHD group and control group (7.9±2.4, 5.8±3.1; 9.2±1.1, 8.2±1.9) (t=3.89, 2.78, P<0.01; effect size=0.70, 0.50), with a greater decrease in the ADHD group. Conclusions The development speed of fine motor ability in children with ADHD aged 6-8 is slow and continues to lag behind normal developmental children. Fine motor development in children with ADHD should be closely monitored, and targeted interventions should be implemented when necessary.

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.

The "student-centered" flipped classroom teaching model can improve students' academic achievement, improve cognition, and cultivate innovation ability. However, it has obvious deficiencies in the integrity and systematization of knowledge as well as education. The traditional teaching concept based on " teacher-centered" has the unique advantages of systematization of knowledge learning and education. Therefore, we integrated the merits of these two different teaching models and introduced the semi-flipped classroom teaching model into the Biochemistry teaching of 2020 stomatology, pharmacy and preventive medicine majors in Cheeloo Medical College, Shandong University, compared with the traditional teaching of 2019 same majors. The data on the improvement of students ' academic achievement and self-cognition were analyzed. The results showed that the students' achievements of the semi-flipped classroom teaching model were better than those of the traditional teaching (P<0. 01). The students' self-cognitions were significantly improved after the implementation of semi-flipped classroom teaching (P<0. 01 or P<0. 05). This study provides a reference for the related teaching and research work in medical colleges.

【Objective】 To quantify the myocardial, hepatic and pancreatic iron overload in β-thalassemia major(β-TM) using MRI T2* technique, and to analyze the relationship of iron deposition between the liver, pancreas and myocardium. 【Methods】 A total of 109 β-TM patients were enrolled in this retrospective study. Clinical and laboratory data were collected and patients were performed 1.5T T2* sequence MR scan on the heart, liver and pancreas. The spearman rank correlation was employed to analyze the relationship between liver T2*, pancreas T2* and myocardium T2*. Wilcoxon rank sum test was used to compare liver T2* values in the two groups: group A(n=32) with patients suffering from both myocardium iron overload(MIO) and liver iron overload(LIO), group B(n=69) with patients suffering from LIO only. Wilcoxon rank sum test was used to compare pancreas T2* values in another two groups: group C(n=34) with patients suffering from both MIO and pancreas iron overload(PIO), group D(n=58) with patients suffering from PIO only. Receiver operating characteristic(ROC) analysis was used to calculate the possibility of using hepatic and pancreatic iron as a predictor of myocardium iron deposition. 【Results】 The median T2* of myocardium, liver and pancreas of the 109 β-TM patients was 27.7(3.2~45.4) ms, 1.8(0.7~18.6) ms, 6.1(1.1~42.9) ms, respectively. With the cut-off level of 20 ms, MIO was detected in 34 cases(31.2%), the youngest one being 7 years old. With the cut-off level of 6.3 ms, LIO was detected in 101 cases(92.7%), the youngest being 5 years old. With the cut-off level of 26 ms, PIO was detected in 92 cases(84.4%), the youngest being 5 years old. Both liver T2*(r= 0.453, P<0.001) and pancreas T2*(r= 0.597, P<0.001) were positively correlated with myocardium T2*. Liver T2* values in group A were lower than those in group B(Z=3.048, P=0.002). Pancreas T2* values in group C were lower than those in group D(Z=6.682, P<0.001). ROC analysis of liver and pancreas R2*(1/T2*) for diagnosing MIO revealed significant distinguishing power of liver R2*(P=0.009) and pancreas R2*(P< 0.001), with area under the curve(AUC) of 0.660, 0.933 and 95% confidence interval of 0.543-0.777, 0.881-0.985, respectively. 【Conclusions】 Liver and pancreas iron overload occurr early and overwhelmingly in β-TM patients. Iron deposits earlier in the liver and pancreas than that in the myocardium. Both hepatic and pancreatic iron level are correlated with myocardial iron level, and can be predictors of the latter.

OBJECTIVE: To study the significance of dishevelled (DVL) proteins in the Wnt signaling pathway in the pathogenesis and prognosis of childhood acute lymphoblastic leukemia (ALL). METHODS: A total of 33 children with new-onset ALL were enrolled as the case group. According to the degree of risk, they were divided into 3 groups: low-risk (n=14), intermediate-risk (n=5) and high-risk (n=14). A total of 29 children with immune thrombocytopenia were enrolled as the control group. At diagnosis and on day 33 of induction therapy, 2 mL bone marrow samples were collected from the case and control groups, and qRT-PCR was used to measure the mRNA expression of DVL1, DVL2 and DVL3 in blood cells of bone marrow. RESULTS: The mRNA expression of DVL1, DVL2 and DVL3 in the case group in the incipient stage was significantly higher than that in the remission stage and the control group (P<0.05). Compared with the control group, the case group had a significant increase in the mRNA expression of DVL2 in the remission stage (P<0.05). The mRNA expression of DVL2 was significantly higher than that of DVL1 and DVL3 in both remission and incipient stages (P<0.05). The high- and intermediate-risk groups had significantly higher mRNA expression of DVL1 and DVL2 than the low-risk group (P<0.05). The mRNA expression of DVL2 was significantly higher than that of DVL1 and DVL3 in the low-, intermediate- and high-risk groups (P<0.05). CONCLUSIONS The change in the expression of DVL, especially DVL2, in the Wnt signal pathway has certain significance in the pathogenesis and prognosis of childhood ALL.
Child ; Dishevelled Proteins ; Humans ; Phosphoproteins ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; Wnt Signaling Pathway

Objective To study the decomposition kinetics of omethoate in blood.Methods The acetonitrile precipitated protein was added into the blood, with the chromatographic column of a Waters BEH C18column (2.1 mm×50 mm, 1.7μm), the mobile phase of 5 mmol/L ammonium acetate aqueous solution-methanol, and the gradient elution with a flow rate of 0.3 mL/min and injection volume of 2μL.With electrospray ionization (ESI) source and positive ion detection, qualitative and quantitative analyses were taken using multi-reaction monitoring mode.Omethoate standard was added into blank human blood to the mass concentrations of 0.78, 1.40, 2.30, 4.50, and 7.20μg/mL, and each mass concentration was preserved at 3 temperatures of-20℃, 4℃, and 20℃, respectively.The content of omethoate was detected at different time points (0, 1, 3, 4, 7, 11, 15, 24, 32, 40, 48, 64, 80, 96, and 120 d).Results Different concentrations of omethoate all showed a descended trend in human blood under different temperature conditions.The decomposition in storage environment of-20℃, 4℃, and 20℃was fit to a one-compartment open model with a first-order kinetic process, which could be expressed as Ct=Coe-αt, with the calculated theoretical values of omethoate concentration close to the measured values.Conclusion All concentrations of omethoate are decomposed in the blood, which vary a lot in different preservation conditions.It is suggested that blood samples should be frozen and detected timely in suspected omethoate poisoning cases.

Objective To establish a combined quality evaluation model of fingerprint and quantitative analysis of multi-components with a single-marker (QAMS) to analyze the total flavonoids from licorice residue by the chemical conversion method; To provide technical support for quality control in production. Methods Total flavonoids of breaking cell wall and enriching were taken as the object of study to establish fingerprint. With liquiritin as internal reference, the relative correction factors of isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid were established respectively, and the contents were determined. Meanwhile, the calculated values were compared with the measured value by external standard method to verify the practicability and stability of QAMS. Results The HPLC fingerprint of total flavonoids from licorice residue was established. 11 common peaks were identified, and 5 common peaks were identified, and the similarity of the 10 extracts was >0.99; the relative error between the calculated results of QAMS and the measured values of the external standard method was <4%; the RSD of relative correction factor calculated by the multiple concentration method was <2%. Conclusion The method is accurate, reliable, specific, and stable, with good repeatability, which can be used for the quality control of total flavonoids from licorice residue.

Objective To establish a method for the simultaneous determination of liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid in licorice extract. Methods Liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid in licorice extract were determined by HPLC dual wavelength spectrophotometry. Symmetry C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase was acetonitrile (A) - 0.085％ phosphoric acid water (B), ingradient elution mode (0–8 min, 81％ B; 8–35 min, 81％→50％ B; 35–60 min, 50％ B) with the flow rate of 1.0 mL/min. The sample size was 10 μL, and column temperature was room temperature. Dual wavelength detection, λ1=237 nm, λ2=254 nm. Results Liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid were linear in the ranges of 0.0408–0.816 μg, 0.0528–1.056 μg, 0.0224–0.448 μg, 0.0212–0.424 μg, and 0.0448–0.896 μg, respectively. The average recovery was 98.69％, 98.31％, 99.10％, 98.55％, and 99.14％, respectively; RSD was 1.39％, 1.29％, 1.78％, 2.14％, and 1.15 ％, respectively. Conclusion The method is accurate, reliable and specific. The results are stable with good repeatability. It can be used for the determine of above 5 components in licorice extract.

The receptor-binding domain(RBD) protein of HCoV-NL63 is a major target in the development of diagnostic assay and vaccine, it has a pivotal role in receptor attachment, viral entry and membrane fusion. In this study, we prepared 2 purified recombinant HCoV-NL63 RBD proteins using in E. coli system and identified the proteins by Western blotting. We first optimized codon and synthesized the RL (232-684aa)coding gene, then amplified the RL or RS(476-616aa) coding gene via PCR using different primers . The RL or RS coding gene was cloned into the pM48 expression vector fused with TrxA tag. The RBD (RL and RS) of HCoV-NL63 were expressed majorly as inclusion body when expressed in E. coli BL21pLys S under different conditions. The expressed products were purified by affinity chromatography then analyzed by SDS-PAGE and Western blotting. Our results showed that the recombinant RBD proteins were maximally expressed at 37 degrees C with 0. 8mM IPTG induction for 4h. RL or RS protein with 95 % purity was obtained and reacted positively with anti-sera from mice immunized with the recombinant vaccinia virus (Tiantan strain) in which HCoV-NL63 RL or RS protein was expressed. In conclusion, the purified recombinant RBD proteins(RL and RS)derived from E. coli were first prepared in China and they might provide a basis for further exploring biological role and vaccine development of HCoV-NL63.
Animals ; Coronavirus Infections ; metabolism ; virology ; Coronavirus NL63, Human ; chemistry ; genetics ; metabolism ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Humans ; Mice ; Mice, Inbred BALB C ; Protein Engineering ; Protein Structure, Tertiary ; Receptors, Virus ; metabolism ; Viral Envelope Proteins ; chemistry ; genetics ; metabolism