Ningxin-Tongyu-Zishen formula alleviates the senescence of granulosa cells on D-galactose-induced premature ovarian insufficiency mice.

Ningxin-Tongyu-Zishen formula (NTZF) is a clinical experience formula for the treatment of premature ovarian insufficiency (POI) in traditional Chinese medicine (TCM), and the potential mechanism is unknown. For in vivo experiments, POI mouse models (C57BL/6 mice), were constructed by subcutaneous injection of D-galactose (D-gal, 200 mg/kg). After treatment of NTZF (10.14, 20.27, 40.54 g/kg;) or estradiol valerate (0.15 mg/kg), ovarian function, oxidative stress (OS) and protein expression of Sirt1/p53 were evaluated. For in vitro experiments, H2O2 (200 µM) was used to treat KGN to construct ovarian granulosa cells (OGCs) cell senescence model. Pretreatment with NTZF (1.06 mg/mL) or p53 inhibitor (Pifithrin-α, 1 µM) was performed before induction of senescence, and further evaluated the cell senescence, OS, mRNA and protein expression of Sirt1/p53. In vivo, NTZF improved ovarian function, alleviated OS and Sirt1/p53 signaling abnormalities in POI mice. In vitro experiments showed that NTZF reduced the level of OS and alleviated the senescence of H2O2-induced KGN. In addition, NTZF activated the protein expression of Sirt1, inhibited the mRNA transcription and protein expression of p53 and p21. Alleviating OGCs senescence and protecting ovarian function through Sirt1/p53 is one of the potential mechanisms of NTZF in the treatment of POI.

Correlation between Spectral CT Parameters and Ki67 Expression in Hepatocellular Carcinoma.

OBJECTIVE: The objective of this study was to analyze the relationship between quantitative parameters of spectral CT and the Ki67 expression index of tumor cells in hepatocellular carcinoma (HCC). METHODS: A total of 19 patients who underwent preoperative spectral CT dual-phase enhancement and who were diagnosed with HCC by postoperative pathology were prospectively selected. Patients with ≥10% Ki67-positive tumor cells formed a high-Ki67 group, and those with <10% Ki67- positive cells formed a low-Ki67 group. The iodine concentrations (ICs) of the lesion and the descending aorta were measured during the arterial and venous phases. Relative iodine concentration (RIC) was calculated thus: RIC=IClesion/ICdescending aorta. CT values of the lesions at 40 and 70 keV were measured during the enhanced arterial and venous phases. The slope of the spectral curve (λ) was calculated thus: λ = (40 keV-70 keV) /(70-30). To compare the differences in quantitative parameters between the high- and low-Ki67 groups, either an independent samples t-test (normal distribution) or a Mann-Whitney U test (non-normal distribution) was used. Receiver operating characteristic curves were used to evaluate the effectiveness of spectral CT parameters in distinguishing between high-Ki67 and low-Ki67 groups. Pearson correlation analysis was used to evaluate the correlation between spectral CT quantitative parameters and Ki67 expression. RESULTS: IC, RIC and λ values for the high-Ki67 group in arterial and venous phases were higher than those for the low-Ki67 group, P < 0.05. IC, RIC, and λ values in the arterial phase were 0.83, 0.89, and 0.75, respectively; in the venous phase, the values of these three parameters were 0.76, 0.77, and 0.69, respectively. IC, RIC, and λ were positively correlated with Ki67 expression in both arterial and venous phases, with a highest correlation of 0.82 for arterial-phase RIC. CONCLUSION: The quantitative parameters of spectral CT in HCC were correlated with Ki67 expression. This finding may make it easier for clinicians to determine whether a tumor is high or low in Ki67 before surgery.

[Effects of Supplementation of Different Amendments on Soil Heavy Metals and Enzyme Activities in Coastal Saline Land].

The supplementation of soil amendments may not only improve the soil physical and chemical properties but also lead to the accumulation of heavy metals in soil. This experiment included six treatments:control (CK), organic manure (OM), polyacrylamide+organic manure (PAM+OM), straw mulching+organic manure (SM+OM), buried straw+organic manure (BS+OM), and bio-organic manure+organic manure (BM+OM) to explore the effects of different soil amendments on heavy metals and soil enzyme activities in coastal saline land and the relationship between them. The results revealed that compared with that in the CK treatment, the contents of soil Cr, Cu, Ni, and Pb exhibited an upward trend after the supplementation of soil amendments, among which the SM+OM and PAM+OM treatments had the most significant effects on the contents of soil Cr and Cu, respectively, whereas the BM+OM treatment had the most significant effects on the contents of soil Ni and Pb. Compared with those in the CK treatment, the activities of soil invertase and urease increased significantly following supplementation of soil amendments, and the BM+OM treatment had the best effect. The alkaline phosphatase activity exhibited a slightly upward trend after the supplementation of soil amendments, whereas the catalase activity did not change significantly. The redundancy analysis revealed that the first two axes cumulatively accounted for 70.3% of the variability in enzyme activities, and the importance of single soil heavy metals on soil enzyme activity was as follows:Ni>Cu>Cr>Pb.

Variation in CFHR3 determines susceptibility to meningococcal disease by controlling factor H concentrations.

Neisseria meningitidis protects itself from complement-mediated killing by binding complement factor H (FH). Previous studies associated susceptibility to meningococcal disease (MD) with variation in CFH, but the causal variants and underlying mechanism remained unknown. Here we attempted to define the association more accurately by sequencing the CFH-CFHR locus and imputing missing genotypes in previously obtained GWAS datasets of MD-affected individuals of European ancestry and matched controls. We identified a CFHR3 SNP that provides protection from MD (rs75703017, p value = 1.1 × 10-16) by decreasing the concentration of FH in the blood (p value = 1.4 × 10-11). We subsequently used dual-luciferase studies and CRISPR gene editing to establish that deletion of rs75703017 increased FH expression in hepatocyte by preventing promotor inhibition. Our data suggest that reduced concentrations of FH in the blood confer protection from MD; with reduced access to FH, N. meningitidis is less able to shield itself from complement-mediated killing.

Intestinal microbiome changes in patients with postpartum depression / 西安交通大学学报(医学版)

【Objective】 To analyze the changes of gut microbes in patients with postpartum depression so as to explore the relationship between postpartum depression and gut microbes. 【Methods】 A total of 60 postpartum subjects were recruited to participate in this study. The depression status of the participants was scored using the Edinburgh Postpartum Depression Scale (EPDS). Those with a score ≥13 were included in the postpartum depression group (PPD group), while those with a score less than 13 were included in the postpartum healthy control group (PPHC group). The feces of these 60 subjects were collected, and the fecal whole genome DNA was extracted for 16S rDNA sequencing. The data of changes in the bacterial diversity between the groups were obtained, and the possible correlation between the changes of intestinal microbes and postpartum depression was analyzed. 【Results】 The number of microorganisms in PPD patients was significantly reduced (P<0.001); the Chao1 index (P<0.001) and ACE index (P<0.001) of α diversity decreased significantly. There were also significant differences in β diversity between the two groups. Analysis of the bacteria in the groups showed that Acetanaerobacterium, Adlercreutzia, Allobaculum, Alloprevotella, Bifidobacterium, Christensenella, Escherichia, Eubacterium, Faecalicatena, Fusobacterium, Haemophilus, Intestinimonas, Lactobacillus, Megamonas, Monoglobumus, Muribaculum, Oscillospira, Paraprevotella, Streptococcus, Raoultibacter, Ruminococcus and Stomatobaculum were significantly enriched in PPHC group. In contrast, Kineothrix, Lachnoclostridium, Acinetobacter, Aquisphaera, Enterococcus, and Mucispirillum were enriched in PPD group. RDA/CCA analysis showed that EPDS was positively correlated with Prevotella, Kineothrix, and Alistipes, but negatively correlated with Lachnospira. 【Conclusion】 This study found that the intestinal flora of patients with postpartum depression was significantly disrupted, and there was a correlation between the intestinal flora and postpartum depression symptom score. Therefore, intestinal microbial markers may contribute to the diagnosis and treatment of patients with postpartum depression.

How confident are pharmacists in providing pharmaceutical care on anticoagulants? A cross-sectional, self-administered questionnaire study in Borneo, Malaysia.

BACKGROUND: Anticoagulants are the cornerstone therapy for the management of venous thromboembolism (VTE) and atrial fibrillation (AF). Pharmacists should be confident and equipped with the skill and updated knowledge in managing anticoagulation therapy. OBJECTIVE: To explore self-reported confidence level of pharmacists, perceived reasons influencing their confidence and socio-demographic associated with high confidence level in the area of anticoagulation. METHODS: A cross-sectional, self-administered questionnaire survey was carried out among fully registered pharmacists who work in selected government hospitals and clinics in Borneo, Malaysia, from January 2019 to February 2020. RESULTS: Overall, responses from 542 fully registered pharmacists were obtained. Proportion of respondents who claimed confident in providing necessary information to patient receiving warfarin (n = 479, 88.3%) was significantly higher (p < 0.001) compared to low molecular weight heparins (n = 317, 58.5%) and direct oral anticoagulants (n = 211, 38.9%). Respondents' perceived reasons that may influence their confidence level include experience in dealing with anticoagulants' cases (n = 469, 86.5%), knowledge on anticoagulants (n = 394, 72.7%) and knowledge on diseases needing anticoagulation therapy (n = 311, 57.4%). Practising as ward pharmacist and "always" dealing with anticoagulants during their practice were the socio-demographic that significantly associated with high confidence level of pharmacist in providing pharmaceutical care on all types of anticoagulants (p < 0.05). CONCLUSION: Pharmacists were found more confident in providing pharmaceutical care on warfarin compared to low molecular weight heparins and direct oral anticoagulants. Continuous educational and training programmes on the use of anticoagulants should be carried out to enhance pharmacists' confidence in supporting patients' care.

Prognostic significance of regional lymphadenectomy in T1b gallbladder cancer: Results from 24 hospitals in China.

BACKGROUND: Whether regional lymphadenectomy (RL) should be routinely performed in patients with T1b gallbladder cancer (GBC) remains a subject of debate. AIM: To investigate whether RL can improve the prognosis of patients with T1b GBC. METHODS: We studied a multicenter cohort of patients with T1b GBC who underwent surgery between 2008 and 2016 at 24 hospitals in 13 provinces in China. The log-rank test and Cox proportional hazards model were used to compare the overall survival (OS) of patients who underwent cholecystectomy (Ch) + RL and those who underwent Ch only. To investigate whether combined hepatectomy (Hep) improved OS in T1b patients, we studied patients who underwent Ch + RL to compare the OS of patients who underwent combined Hep and patients who did not. RESULTS: Of the 121 patients (aged 61.9 ± 10.1 years), 77 (63.6%) underwent Ch + RL, and 44 (36.4%) underwent Ch only. Seven (9.1%) patients in the Ch + RL group had lymph node metastasis. The 5-year OS rate was significantly higher in the Ch + RL group than in the Ch group (76.3% vs 56.8%, P = 0.036). Multivariate analysis showed that Ch + RL was significantly associated with improved OS (hazard ratio: 0.51; 95% confidence interval: 0.26-0.99). Among the 77 patients who underwent Ch + RL, no survival improvement was found in patients who underwent combined Hep (5-year OS rate: 79.5% for combined Hep and 76.1% for no Hep; P = 0.50). CONCLUSION: T1b GBC patients who underwent Ch + RL had a better prognosis than those who underwent Ch. Hep + Ch showed no improvement in prognosis in T1b GBC patients. Although recommended by both the National Comprehensive Cancer Network and Chinese Medical Association guidelines, RL was only performed in 63.6% of T1b GBC patients. Routine Ch + RL should be advised in T1b GBC.

Correlation of serum inflammatory factors with clinical symptoms and cognitive functions in schizophrenia / 西安交通大学学报(医学版)

【Objective】 To study the correlation of clinical symptoms and cognitive functions with serum inflammatory factors in schizophrenia. 【Methods】 A total of 42 SCz patients (case group) and 47 healthy controls (control group) were enrolled in this study. We used enzyme-linked immunosorbent assay (ELSA) to determine six inflammatory factors in serum. PANSS was used to assess clinical symptoms and MCCB was used to assess the patients’ cognitive functions. 【Results】 ① Inflammatory factors: The serum levels of IL-1β, IL-4, IL-6 and IL-8 were significantly higher in case group than in control group (P<0.01). ② Cognitive functions: The scores of Trail Making Test, Hopkins Verbal Learning Test, Symbol Coding, Spatial Span, Brief Visuospatial Memory Test, Assessment Battery-Mazes, Category Fluency and Test-Managing Emotions of case group were significantly lower than those of control group (P<0.05). ③ Correlation between serum inflammatory factors and clinical symptoms: There was no correlation between serum inflammatory factors and psychiatric symptoms in schizophrenia. ④ Correlation between serum inflammatory factors and cognitive functions: The levels of IL-6 (rs=-0.33, P<0.05) and IL-8 (rs=-0.50, P<0.01) in case group were significantly negatively correlated with the scores of Space Scan test. 【Conclusion】 Patients with schizophrenia are presented with immune dysfunction, and the latter is correlated with cognitive impairments.

Subtrochanteric Osteotomy in Direct Anterior Approach Total Hip Arthroplasty.

Subtrochanteric osteotomy of the femur (STO) is a valuable corrective procedure in hip surgeries. However, STO in traditional posterolateral approach usually encounters complications such as postoperative dislocation, bone non-union, and prosthesis failure. Some relevant pathologies and mechanisms have been identified, but there is sparse evidence for verification. The aim of this video in orthopaedic technique is to test our hypothesis of STO in direct anterior approach to total hip arthroplasty in a complicated hip surgery, and to further illustrate the rationality, reproducibility, and superiority of STO in this minimally invasive and enhanced-recovery approach by presenting a standardized and systemic protocol, as well as operational pearls and pitfalls.

Direct Anterior Approach in Crowe Type III-IV Developmental Dysplasia of the Hip: Surgical Technique and 2 years Follow-up from Southwest China.

OBJECTIVES: To summarize our pioneering surgical practice and clinical outcome of Crowe type III-IV developmental dysplasia of the hip (DDH) with a direct anterior approach total hip arthroplasty in a single teaching hospital in Southwest China. METHODS: Fourteen patients (15 hips) diagnosed with Crowe type III-IV developmental dysplasia of the hip were involved in this single-center retrospective study between 2016 and 2018. A comprehensive surgical procedure, including preoperative planning and algorithms for leg length equalization, intraoperative stepwise soft tissue release, bone defect reconstruction, and an innovative subtrochanteric osteotomy, was described. Furthermore, advancements in intraoperative CT guidance, computer navigation, and nerve monitoring were available for specific demands. The short-term clinical outcome was evaluated at the endpoint of follow-up by three patient-reported functional scales (Harris, WOMAC, and SF-12 scores), and objective data collected at the clinic, including functional recovery (muscle strength of hip flexor and abductor, correction of the pelvic tilt, leg length discrepancy, and limp), radiographic analysis, and complication occurrence. RESULTS: The intraoperative variables were carefully recorded. The mean operating times in Crowe type III and IV groups were 115.8 min and 156.2 min, and the median blood loss volumes were 520.5 mL and 810.2 mL, respectively. The general changes in the Harris, SF-12, and WOMAC scores of the two groups were 46.2, 8.7 and 134.3, respectively, at a mean follow-up of 25.4 months. Enhanced recovery of hip abductor muscle strength was identified in 85.7% of the population at the third postoperative month. The equalization of leg length and correction of the pelvic tile were observed at the sixth postoperative month, with a 36-mm decrease in leg length discrepancy. No radiographic evidence of the loosening or migration of the components was observed. A self-innovated subtrochanteric shortening osteotomy was performed in five patients, and they healed after 6 months. Specific complications included two cases of distal femoral cracks and one case of a periprosthetic fracture needing internal fixation. Two patients received a late iliotibial band release at the 3rd month postoperatively due to progressive genu valgum. No records of infection, dislocation, nerve palsy, bone non-union, or revision surgery were identified. DISCUSSION: The direct anterior approach total hip arthroplasty showed potential advantages, including optimum component positioning, improved hip stability, steerable complication rate, and enhanced functional recovery with Crowe type III-IV DDH. The short-term outcome is comparable to that of the traditional posterolateral approach.

Molecular hydrogen produced by elemental magnesium inhibits rumen fermentation and enhances methanogenesis in dairy cows.

Hydrogen is a key metabolite that connects microbial fermentation and methanogenesis in the rumen. This study was to investigate the effects of elevated H2 produced by elemental Mg on rumen fermentation and methanogenesis in dairy cows. Four nonlactating Chinese Holstein dairy cows were employed for this experiment in a replicated crossover design. The 2 dietary treatments included a basal diet supplemented with Mg(OH)2 (14.5 g/kg of feed dry matter) or elemental Mg (6.00 g/kg of feed dry matter). When compared with Mg(OH)2 treatment, cows fed diet with elemental Mg had similar rumen Mg2+ concentration, but higher rumen dissolved H2 and methane concentrations at 2.5 h after morning feeding. Also, elemental Mg supplementation decreased feed digestibility, rumen volatile fatty acid concentration, and relative abundance of group Ruminococcaceae_UCG-014, genus Bifidobacterium, and group Mollicutes_RF9, increased acetate to propionate ratio, succinate concentration, and abundance of family Christensenellaceae. Elemental Mg supplementation increased enteric CH4 emission, altered methanogen community with increased abundance of order Methanomassiliicoccales, 16S ribosomal RNA gene copies of methanogens, and order Methanobacteriales. In summary, the pulse of elevated dissolved H2 after feeding produced by elemental Mg inhibited rumen fermentation and feed digestibility by decreasing the abundance of carbohydrate-degrading bacteria, promoted H2 incorporation into succinate by increasing family Christensenellaceae and genus Bacteroidales_BS11, and increased H2 utilization for methanogenesis by favoring growth of methanogens.

cnvCapSeq: detecting copy number variation in long-range targeted resequencing data.

Targeted resequencing technologies have allowed for efficient and cost-effective detection of genomic variants in specific regions of interest. Although capture sequencing has been primarily used for investigating single nucleotide variants and indels, it has the potential to elucidate a broader spectrum of genetic variation, including copy number variants (CNVs). Various methods exist for detecting CNV in whole-genome and exome sequencing datasets. However, no algorithms have been specifically designed for contiguous target sequencing, despite its increasing importance in clinical and research applications. We have developed cnvCapSeq, a novel method for accurate and sensitive CNV discovery and genotyping in long-range targeted resequencing. cnvCapSeq was benchmarked using a simulated contiguous capture sequencing dataset comprising 21 genomic loci of various lengths. cnvCapSeq was shown to outperform the best existing exome CNV method by a wide margin both in terms of sensitivity (92.0 versus 48.3%) and specificity (99.8 versus 70.5%). We also applied cnvCapSeq to a real capture sequencing cohort comprising a contiguous 358 kb region that contains the Complement Factor H gene cluster. In this dataset, cnvCapSeq identified 41 samples with CNV, including two with duplications, with a genotyping accuracy of 99%, as ascertained by quantitative real-time PCR.

[Killing activity in DC and CIK co-culture against hepatocarcinoma cells].

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.

Killing activity in DC and CIK co-culture against hepatocarcinoma cells / 中国实验血液学杂志

This study was aimed to investigate the proliferation activities and phenotype changes of DC, CIK and DC-CIK, and their cytotoxicity against hepatocarcinoma cells in co-culture of DC with CIK. Peripheral blood mononuclear cells (PBMNC) were isolated from healthy adult donors. After incubation of PBMNC for 2 hours, DCs were induced from adherent cells by some cytokines and CIKs were generated from non-adherent cells. Mature DCs were harvested after incubation for 9 days, and then were co-cultured with CIK at ratio of 1:5 for 3 days. The cytotoxicity activity against SMMC-7721 hepatocellular carcinoma cell line was detected by MTT assay. The results showed that CIK cells were able to lyse SMMC-7721 hepatocellular carcinoma cells at low ratios of effector to target. This effect was significantly enhanced by co-culture with DCs. It is concluded that CIK cells have high lytic activity against 7721 hepatocellular carcinoma cell line, which can be enhanced by co-culture with DC. DC-CIK cells are highly effective immune cells.

[Effect of dendritic cells co-cultured with cytokine induced killer cells on cytotoxicity against drug resistant K562 cells].

OBJECTIVE: To study the effect of dendritic cells (DC) co-cultured with cytokine induced killer (CIK) cells on cytotoxicity against K562 and K562 drug-resistant (K562/ADM) cells. METHODS: Peripheral blood mononuclear cells (MNC) isolated from healthy adult donors were induced to obtain CIK cells and DC respectively and then these two kinds of cells were co-cultured. The cytotoxicity of the co-cultured cells against K562 and K562/ADM cells was measured with MTT assay. RESULTS: The cytotoxicity CIK cells alone to K562 and K562/ADM cells was (20.0 +/- 1.2)% - (61.1 +/- 2.2)% and (17.5 +/- 2.1)% - (45.2 +/- 3.3)% respectively at low effector to target ratios (2.5 - 20.0). This effect was significantly enhanced by co-culturing with DCs being (25.2 +/- 2.3)% - (70.9 +/- 4.1)% and (22.4 +/- 2.7)% - (62.3 +/- 5.0)%. CONCLUSION: CIK cells showed high cytotoxicity against K562 and K562/ADM cells and the activity could be enhanced by co-culturing with DC.

Effect of dendritic cells co-cultured with cytokine induced killer cells on cytotoxicity against drug resistant K562 cells / 中华血液学杂志

<p><b>OBJECTIVE</b>To study the effect of dendritic cells (DC) co-cultured with cytokine induced killer (CIK) cells on cytotoxicity against K562 and K562 drug-resistant (K562/ADM) cells.</p><p><b>METHODS</b>Peripheral blood mononuclear cells (MNC) isolated from healthy adult donors were induced to obtain CIK cells and DC respectively and then these two kinds of cells were co-cultured. The cytotoxicity of the co-cultured cells against K562 and K562/ADM cells was measured with MTT assay.</p><p><b>RESULTS</b>The cytotoxicity CIK cells alone to K562 and K562/ADM cells was (20.0 +/- 1.2)% - (61.1 +/- 2.2)% and (17.5 +/- 2.1)% - (45.2 +/- 3.3)% respectively at low effector to target ratios (2.5 - 20.0). This effect was significantly enhanced by co-culturing with DCs being (25.2 +/- 2.3)% - (70.9 +/- 4.1)% and (22.4 +/- 2.7)% - (62.3 +/- 5.0)%.</p><p><b>CONCLUSION</b>CIK cells showed high cytotoxicity against K562 and K562/ADM cells and the activity could be enhanced by co-culturing with DC.</p>

[Effects of oleate on ATP binding cassette transporter A1 expression and cholesterol efflux in THP-1 macrophage-derived foam cells].

To study the effect of oleate on ATP binding cassette transporter A1 (ABCA1) expression and cholesterol efflux in THP-1 macrophage-derived foam cells, after exposure of the cultured THP-1 macrophage-derived foam cells to oleate for different time, cholesterol efflux was determined by FJ-2107P type liquid scintillator. ABCA1 mRNA and its protein level were determined by RT-PCR and Western blot, respectively. The mean ABCA1 fluorescence intensity of THP-1 macrophage-derived foam cells was detected by flow cytometry. The results showed that oleate markedly inhibited ABCA1-mediated cholesterol efflux from THP-1 macrophage-derived foam cells. This was accompanied by a reduction in the membrane content of ABCA1. Oleate did not alter ABCA1 mRNA abundance, indicating that decreased ABCA1 transcription, enhanced mRNA decay, or impaired translation efficiency did not account for these inhibitory effects. Oleate, however, increased ABCA1 turnover when protein synthesis was blocked by cycloheximide. Oleate reduces cholesterol efflux and the level of ABCA1 protein in THP-1 macrophage-derived foam cells.