Objective:To explore the prognostic value of serum free light chain in chronic lymphocytic leukemia patients.Methods:Retrospective cohort study was conducted. One hundred and fifty-six newly diagnosed chronic lymphocytic leukemia(CLL) patients in the first affiliated hospital of Nanjing Medical University from January 2016 to December 2020 were included in the retrospective analysis. Among them, there were 106 males and 50 females, with a median age of 60.7 (53.4, 66.0) years old.Serum sample was collected, serum free light chains were detected, and patients were divided into a treatment group (106 cases) and a follow-up group (50 cases) based on the presence of treatment indications.The threshold of serum free light chain(sFLC) was defined by the reference range of the instruction manual and ROC curve. Three indicators were used, including sFLCR, sFLC(κ+λ) and sFLC(κ-λ). Patients were divided into normal sFLCR group ( n=61)and abnormal group( n=95), as well as sFLC (κ+λ) low value group ( n=88) and high value group ( n=68), and sFLC (κ-λ) low value group ( n=64) and high value group ( n=92).The abnormal group and high value group were enrolled as the experimental group, while the normal group and low value group were enrolled as control group. Chi-square test and Fisher′s exact test were used to compare the clinical data, cytogenetics, and molecular biology characteristics of patients in two groups, Kaplan-Meier method was used to analyze the median treatment-free survival (TFS) of the patients, and Cox regression was used to screen the prognostic factors of the patients. Results:The proportion of Rai stage Ⅰ-Ⅳ ( χ2=8.16, P<0.05 and χ2=7.63, P<0.05 and χ2=5.45, P<0.05), Binet stage B-C( χ2=4.11, P<0.05 and χ2=9.43, P<0.05 and χ2=7.34, P<0.05), β 2-microglobulin>3.5 mg/L( χ2=5.13, P<0.05 and χ2=18.3, P<0.05 and χ2=12, P<0.05), ATM gene mutation rate( χ2=6.21, P<0.05 and χ2=4.88, P<0.05 and χ2=5.19, P<0.05), and immunoglobulin heavy chain variable region (IGHV) mutation free rate ( χ2=18.9, P<0.05 and χ2=24.6, P<0.05 and χ2=10.4, P<0.05)in the experimental group were significantly higher than that in control group 1 ( P<0.05). Multivariate analysis indicated that sFLC(κ+λ) ( HR=1.615,95% CI 1.012-2.576, P=0.044), β 2-microglobulin>3.5 mg/L( HR=2.103,95% CI 1.356-3.262, P=0.001) and TP53 deletion and/or mutation( HR=1.892,95% CI 1.082-3.308, P=0.025) were independent prognostic factors affecting the patients time to first treatment(TFT). Conclusions:Serum free light chains can predict the risk of early treatment and have good prognostic significance in newly diagnosed CLL patients.

Objective: To investigate the effects and mechanisms of Astragalus Polysacharin(APS) on the proliferation and metastasis of breast cancer cells by regulating miR-107 and miR-346-mediated macrophage polarization in breast cancer-derived exosomes. Methods: Forty 8-week-old female BALB/c mice were selected and breast cancer xenograft models and 4T1 transplanted tumor models were established. The mice were divided into the control group and the APS group. The APS group mice received daily intragastric administration of APS for 25 days, while the control group mice were given the same amount of normal saline. After all treatments were completed, the mice were euthanized, and tumor tissues were isolated. Western blot and flow cytometry were used to detect the expressions of proliferating cell nuclear antigen(PCNA), Ki-67, CD206, CD163, inducible nitric-oxide synthase(iNOS), and CD86. The apoptosis of single-cell suspensions in tumor tissues was analyzed. Human breast cancer cell line MDA-MB-231 was cultured and stimulated with APS, and exosomes from the cell culture medium were collected. The proliferation, migration, and invasion of cells were detected by CCK-8 assay, scratch assay, permeability chamber cell invasion assay, and qRT-PCR. Differentially expressed genes were screened by bioinformatics. Results : By measuring the expressions of molecules related to breast cancer cell proliferation and metastasis, it was shown that APS treatment reduced the expressions of proliferation-related proteins(PCNA and Ki-67) and metastasis-related proteins(Vimentin) in MDA-MB-231 xenograft tumor tissues; and the polarization of tumor-associated macrophages was observed. APS treatment of 4T1 transplanted tumor tissues could reduce the number of M2 macrophages and increase the number of M1 macrophages, resulting in a decrease in the ratio of M2/M1 macrophages and an increase in cell apoptosis in 4T1 transplanted tumor tissues. The expressions of related proteins iNOS and CD86 increased, and CD206 and CD163 decreased. After APS treatment, the exosomes produced by MDA-MB-231 reduced the polarization of M2 macrophages and affected the expressions of miR-107 and miR-346. Conclusion APS inhibits the polarization of M2 macrophages by regulating the expression of miR-107 or miR-346 in breast cancer cell-derived exosomes, ultimately inhibiting the proliferation and metastasis of breast cancer cells.

Objective To screen the anti-Streptococcus pneumoniae(Spn)active ingredients in vitro from different po-lar parts of Pilea peltata,and to examine the combined antibacterial effect of the active ingredient and amoxicillin(AMX).Methods A 96-well plate microdilution method was used to determine the minimum inhibitory concentration(MIC)of different polar parts;the most active polar part was separated by preparative high-performance liquid chromatography,and the active ingre-dients were identified using spectral technology.The fractional inhibitory concentration(FIC)of active ingredients and AMX was determined by the 96-well plate chessboard microdilution method.The crystal violet method was used to investigate the effect of ac-tive ingredients on Spn biofilm.The effect of active ingredients on the appearance and morphology of Spn was investigated under the electron microscope.Results The MICs of the petroleum ether part,chloroform part,ethyl acetate part,n-butanol part,and water part were 1.000,1.000,0.500,1.000,and 2.000 mg·mL-1 respectively,among which the ethyl acetate part had the stron-gest antibacterial activity.Three compounds were isolated from ethyl acetate,namely 5,7,3',4'-tetramethoxyflavone 1,8-O-(p-coumaroyl)-1(10)E,4(5)E-humuladien-8-ol 2 and 1-O-p-coumaroyl copaborneol 3.These compounds were all isolated for the first time from Pilea peltate,their MICs against Spn were 200.000,50.000,and 25.000 μg·mL-1 respectively,and the compound 3 had the strongest antibacterial activity;the FIC value of AMX and compound 3 was 0.50,which had a synergistic antibacterial effect on Spn.Both AMX and compound 3 had inhibitory effects on Spn biofilm,but the biofilm inhibition rate of compound 3(59.10±1.04％)was significantly lower than AMX(87.38±0.84)％(P＜0.01);Moreover,there was no significant difference in biofilm inhibition rate between the combination of the two and AMX(P＞0.05).The scanning electron microscope results showed that the bacterial cells in the compound 3 group had a smooth surface but varying degrees of depression.The surface of the bacteri-al cells in the AMX group and the AMX combined compound 3 group showed severe swelling and rupture.Conclusions Fla-vonoids and sesquiterpenoids are both the anti-Spn active components of Pilea peltate.Among them,sesquiterpenoids have more potent antibacterial activity,and their antibacterial action mechanism is related to inhibiting bacterial biofilms.Compound 3 and AMX have a synergistic antibacterial effect on Spn,but its mechanism of action is not by enhancing biofilm inhibition;although compound 3 cannot destroy the cell wall of Spn,it still has a negative impact on the appearance of the bacteria.

Objective To screen the anti-Streptococcus pneumoniae(Spn)active ingredients in vitro from different po-lar parts of Pilea peltata,and to examine the combined antibacterial effect of the active ingredient and amoxicillin(AMX).Methods A 96-well plate microdilution method was used to determine the minimum inhibitory concentration(MIC)of different polar parts;the most active polar part was separated by preparative high-performance liquid chromatography,and the active ingre-dients were identified using spectral technology.The fractional inhibitory concentration(FIC)of active ingredients and AMX was determined by the 96-well plate chessboard microdilution method.The crystal violet method was used to investigate the effect of ac-tive ingredients on Spn biofilm.The effect of active ingredients on the appearance and morphology of Spn was investigated under the electron microscope.Results The MICs of the petroleum ether part,chloroform part,ethyl acetate part,n-butanol part,and water part were 1.000,1.000,0.500,1.000,and 2.000 mg·mL-1 respectively,among which the ethyl acetate part had the stron-gest antibacterial activity.Three compounds were isolated from ethyl acetate,namely 5,7,3',4'-tetramethoxyflavone 1,8-O-(p-coumaroyl)-1(10)E,4(5)E-humuladien-8-ol 2 and 1-O-p-coumaroyl copaborneol 3.These compounds were all isolated for the first time from Pilea peltate,their MICs against Spn were 200.000,50.000,and 25.000 μg·mL-1 respectively,and the compound 3 had the strongest antibacterial activity;the FIC value of AMX and compound 3 was 0.50,which had a synergistic antibacterial effect on Spn.Both AMX and compound 3 had inhibitory effects on Spn biofilm,but the biofilm inhibition rate of compound 3(59.10±1.04％)was significantly lower than AMX(87.38±0.84)％(P＜0.01);Moreover,there was no significant difference in biofilm inhibition rate between the combination of the two and AMX(P＞0.05).The scanning electron microscope results showed that the bacterial cells in the compound 3 group had a smooth surface but varying degrees of depression.The surface of the bacteri-al cells in the AMX group and the AMX combined compound 3 group showed severe swelling and rupture.Conclusions Fla-vonoids and sesquiterpenoids are both the anti-Spn active components of Pilea peltate.Among them,sesquiterpenoids have more potent antibacterial activity,and their antibacterial action mechanism is related to inhibiting bacterial biofilms.Compound 3 and AMX have a synergistic antibacterial effect on Spn,but its mechanism of action is not by enhancing biofilm inhibition;although compound 3 cannot destroy the cell wall of Spn,it still has a negative impact on the appearance of the bacteria.

Objective:To explore the prognostic value of serum free light chain in chronic lymphocytic leukemia patients.Methods:Retrospective cohort study was conducted. One hundred and fifty-six newly diagnosed chronic lymphocytic leukemia(CLL) patients in the first affiliated hospital of Nanjing Medical University from January 2016 to December 2020 were included in the retrospective analysis. Among them, there were 106 males and 50 females, with a median age of 60.7 (53.4, 66.0) years old.Serum sample was collected, serum free light chains were detected, and patients were divided into a treatment group (106 cases) and a follow-up group (50 cases) based on the presence of treatment indications.The threshold of serum free light chain(sFLC) was defined by the reference range of the instruction manual and ROC curve. Three indicators were used, including sFLCR, sFLC(κ+λ) and sFLC(κ-λ). Patients were divided into normal sFLCR group ( n=61)and abnormal group( n=95), as well as sFLC (κ+λ) low value group ( n=88) and high value group ( n=68), and sFLC (κ-λ) low value group ( n=64) and high value group ( n=92).The abnormal group and high value group were enrolled as the experimental group, while the normal group and low value group were enrolled as control group. Chi-square test and Fisher′s exact test were used to compare the clinical data, cytogenetics, and molecular biology characteristics of patients in two groups, Kaplan-Meier method was used to analyze the median treatment-free survival (TFS) of the patients, and Cox regression was used to screen the prognostic factors of the patients. Results:The proportion of Rai stage Ⅰ-Ⅳ ( χ2=8.16, P<0.05 and χ2=7.63, P<0.05 and χ2=5.45, P<0.05), Binet stage B-C( χ2=4.11, P<0.05 and χ2=9.43, P<0.05 and χ2=7.34, P<0.05), β 2-microglobulin>3.5 mg/L( χ2=5.13, P<0.05 and χ2=18.3, P<0.05 and χ2=12, P<0.05), ATM gene mutation rate( χ2=6.21, P<0.05 and χ2=4.88, P<0.05 and χ2=5.19, P<0.05), and immunoglobulin heavy chain variable region (IGHV) mutation free rate ( χ2=18.9, P<0.05 and χ2=24.6, P<0.05 and χ2=10.4, P<0.05)in the experimental group were significantly higher than that in control group 1 ( P<0.05). Multivariate analysis indicated that sFLC(κ+λ) ( HR=1.615,95% CI 1.012-2.576, P=0.044), β 2-microglobulin>3.5 mg/L( HR=2.103,95% CI 1.356-3.262, P=0.001) and TP53 deletion and/or mutation( HR=1.892,95% CI 1.082-3.308, P=0.025) were independent prognostic factors affecting the patients time to first treatment(TFT). Conclusions:Serum free light chains can predict the risk of early treatment and have good prognostic significance in newly diagnosed CLL patients.

Objective:To investigate the role and mechanism of vitamin D(VD)in renal ischemia-reperfusion(I/R group)injury.Methods:Forty-eight male C57BL/6J mice were divided into four groups:Sham operation group(Sham group),active VD analog alfacalcidol treatment group(VD group),renal ischemia-reperfusion injury group(I/R group)and alfacalcidol treated I/R group(I/R+VD group).After 24 h renal I/R injury,mice in each group were sacrificed,and peripheral blood was collected to detect the levels of blood urea nitrogen(BUN),serum creatinine(SCr)and inflammatory factors IL-1β,IL-18 and tumor necrosis factor-α(TNF-α).The renal tissues of mice in each group were collected,the apoptosis level of renal tissues was detected by TUNEL staining,and the positive expression rate of NLRP3 was detected by IHC.Western blot was used to detect the NLRP3 inflammasome associated factors NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β and NF-κB p65,IκBα in renal tissues of mice in each group.Results:Com-pared with Sham group,there was no significant difference in serum BUN,SCr,IL-1β,IL-18 and TNF-α levels in VD group(P>0.05),significantly increased in I/R group and I/R+VD group(P<0.05).Compared with I/R group,BUN,SCr,IL-1β,IL-18 and TNF-α were significantly decreased in I/R+VD group(P<0.05).Compared with Sham group,the positive rate of TUNEL and NLRP3 expression in renal tissues of mice in VD group had no significant difference(P>0.05),the positive rate of TUNEL and NLRP3 expres-sion in renal tissues of mice in I/R group and I/R+VD group were significantly increased(P<0.01),while the positive rate of TUNEL and NLRP3 expression in I/R+VD group were significantly decreased(P<0.05).Western blot results showed that compared with Sham group,there was no significant difference in protein expression levels of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β,NF-κB P65,IκBα in VD group(P>0.05).The protein expression of IκBα in I/R group and I/R+VD group was significantly decreased(P<0.05),while the other protein expression levels were significantly increased(P<0.05).Compared with the I/R group,the expression levels of NLRP3,GSDMD-N,cleaved Caspase-1,IL-1β and NF-κB p65 were significantly decreased in the I/R+VD group(P<0.05),while the expression level of IκBα was significantly decreased(P<0.05).Conclusion:VD plays a protective role in I/R injury by inhibiting NF-κB mediated activation of NLRP3 inflammasome.

Objective:To explore the clinical features and genetic etiology for a child with developmental delay, impaired growth, facial dysmorphism, and axonal neuropathy (DIGFAN).Methods:A child who was admitted to the Second Affiliated Hospital of Guangxi Medical University on March 22, 2021 was selected the study subject. Clinical data of the child was collected. Following extraction of genomic DNA, the child and his parents were subjected to whole exome sequencing (WES), and candidate variant was verified by Sanger sequencing and bioinformatic analysis.Results:The child, a 10-year-and-9-month-old boy, had manifested with short stature, intellectual disability, delayed speech, motor and language development, and facial dysmorphism. WES and Sanger sequencing revealed that he has harbored a novel de novo c. 800T>C (p.Leu267Pro) variant of the MORC2 gene. The Leucine at position 267, which is highly conserved among various species, is located in the S5 domain of ribosome protein in the ATPase binding region of MORC2. And the Leu267Pro may affect the function of MORC2 by altering the spatial conformation and activity of ATPase. Based on the guidelines from the American College of Medical Genetics and Genomics, the c. 800T>C variant was classified as likely pathogenic (PS2+ PM2_Supporting+ PP2+ PP3). Conclusion:The MORC2: c. 800T>C (p.Leu267Pro) variant probably underlay the pathogenesis of DIGFAN syndrome in this child.

Objective:To understand the attitude, practice status and influencing factors of obstetric medical staff on sexual health care during pregnancy, and to provide reference for carrying out sexual education and training during pregnancy.Methods:A self-designed questionnaire on attitude and practice of obstetric medical staff towards sexual health care during pregnancy was used to investigate 462 obstetric medical staff in Guangdong Province from July to August, 2021,and the influencing factors were analyzed.Results:The attitude score of sexual health care during pregnancy among obstetric medical staff was (29.87 ± 5.96) points, and the practice score was (13.61 ± 1.23) points. Profession and hospital level affected obstetric medical staff′s attitude towards sexual health care during pregnancy ( P<0.05); profession, title and hospital level affected obstetric medical staff′s practice in providing pregnancy sexual health care ( P<0.05). Conclusions:Obstetric medical staff have a negative attitude towards sexual health care during pregnancy, and their active sexual health care behaviors need to be improved. Medical schools and work units need to strengthen the training and management of obstetric-related and reproductive health knowledge and skills to promote the effectiveness of pregnancy sexual education implementation and promotion.

Hepatic ischemia/reperfusion injury (HIRI) is a serious complication that occurs following shock and/or liver surgery. Gut microbiota and their metabolites are key upstream modulators of development of liver injury. Herein, we investigated the potential contribution of gut microbes to HIRI. Ischemia/reperfusion surgery was performed to establish a murine model of HIRI. 16S rRNA gene sequencing and metabolomics were used for microbial analysis. Transcriptomics and proteomics analysis were employed to study the host cell responses. Our results establish HIRI was significantly increased when surgery occurred in the evening (ZT12, 20:00) when compared with the morning (ZT0, 08:00); however, antibiotic pretreatment reduced this diurnal variation. The abundance of a microbial metabolite 3,4-dihydroxyphenylpropionic acid was significantly higher in ZT0 when compared with ZT12 in the gut and this compound significantly protected mice against HIRI. Furthermore, 3,4-dihydroxyphenylpropionic acid suppressed the macrophage pro-inflammatory response in vivo and in vitro. This metabolite inhibits histone deacetylase activity by reducing its phosphorylation. Histone deacetylase inhibition suppressed macrophage pro-inflammatory activation and diminished the diurnal variation of HIRI. Our findings revealed a novel protective microbial metabolite against HIRI in mice. The potential underlying mechanism was at least in part, via 3,4-dihydroxyphenylpropionic acid-dependent immune regulation and histone deacetylase (HDAC) inhibition in macrophages.

Objective:To explore the protective effect of emodin on D-galactosamine (D-GalN)/lipopolysaccharide (LPS)-induced acute liver injury and its mechanism.Methods:A total of 40 male BALB/c mice were randomly (random number) divided into 5 groups ( n=8 in each group): the control group, the emodin group, the D-GalN/LPS group, the emodin+D-GalN/LPS group and the 3-MA+emodin+D-GalN/LPS group. D-GalN (700 mg/kg) and LPS (10 μg/kg) were intraperitoneally injected to induce acute liver injury in mice. Autophagy inhibitor 3-MA (15 mg/kg) and/or emodin (20 mg/kg) were intraperitoneally injected 30 min before the liver injury model. The animals were sacrificed under anaesthesia 6 h after D-GalN/LPS challenge, blood samples and liver tissues were collected. The levels of alanineaminotransferase (ALT) and aspartateaminotransferase (AST) in serum, and myeloperoxidase (MPO) activity of liver tissues were determined by colorimetric quantitative method; the levels of tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6) were measured by ELISA; the expression of LC3-II and Beclin 1 in the liver tissues were evaluated by Western blot; the pathological changes of liver was evaluated by HE staining. Animal survival rate was also analyzed. The one-way ANOVA was use to compare quantitative data, SNK- q test was used for pairwise comparison between two groups, and Games-Howell test was used when homogeneity of variance were not met. Results:Compared with the control group, the levels of ALT, AST, TNF-α, IL-6 and MPO activity [(2 476.80 ± 263.14) U/L, (271.71 ± 47.15) U/L, (537.92 ± 89.35) pg/mL, (169.74 ± 25.52) pg/mL, and (1.37 ± 0.22) U/mg] were obviously increased in the D-GalN/LPS group ( P<0.05). Compared with the D-GalN/LPS group, the levels of ALT, AST, TNF-α, IL-6 and MPO activity [(1 248.01 ± 380.70) U/L, (142.59 ± 34.63) U/L, (288.91 ± 67.21) pg/mL, (61.83 ± 13.64) pg/mL, and (0.80 ± 0.21) U/mg] were obviously decreased in the emodin+ D-GalN/LPS group ( P<0.05). Compared with the D-GalN/LPS group, the histopathological abnormalities in liver tissue were significantly alleviated and the survival rate of mice was improved in the emodin+ D-GalN/LPS group. Compared with the control group, the expression of LC3-II and Beclin1 was decreased in the liver tissue in the D-GalN/LPS group, while compared with the D-GalN/LPS group, the expression of LC3-II and Beclin1 was increased in the emodin+ D-GalN/LPS group. With co-administration of 3-MA, the protective effects of emodin in acute liver injury were reversed, the levels of AST, ALT, TNF-α, IL-6, and MPO [(2 398.78 ± 233.57) U/L, (242.79 ± 43.46) U/L, (505.07 ± 67.89) pg/mL, (151.46 ± 14.11) pg/mL, and (1.27 ± 0.15) U/mg] were increased, and the pathological damage of liver tissue was aggravated. Conclusions:Emodin alleviates D-GalN/LPS-induced acute liver injury in mice, which may be related to the activation of protein LC3-II, Beclin1 and restored autophagy.