Objective To investigate the infiltration of peripheral monocyte in the hippocampal CA3 area in neuralgia mice at different time points and explore the effects of the infiltration on neuralgia and the neuralgia-induced anxiety-like behavior in mice.Methods The healthy male C57 mice were randomly divided into four groups:sham,sciatic nerve branch selective injury(SNI)model(SNI),CCR2 inhibitor RS102895(SNI+RS102895)and microglial inhibitor minocycline(MC)(SNI+MC)groups.Both the sham and SNI groups were further divided into 7 days,14 days and 18 days groups,and the SNI+RS102895 and SNI+MC groups were sampled on the 18th day.Neuralgia was induced by SNI,and mechanical hyperalgesia was assessed by paw withdrawal threshold(PWTs)at different time points.Elevated plus maze(EPM)and open field test(OFT)were performed respectively two days and one day before sacrifice.Immunofluorescence was used to observe the expressions of leukocyte differentiation antigen 45(CD45)and the co-expression with microglial markers ionized calcium binding adaptor molecule-1(IBA-1),transmembrane protein 119(TMEM119),astrocyte marker glial fibrillary acidic protein(GFAP),and neuronal marker neuronal nuclei(NeuN)in the hippocampal CA3.The percentage of monocytes in the whole brain of 14 days SNI mice was determined by flow cytometry.Minocycline at 90 mg/(kg·d),RS 102895 at 5 mg/(kg·d)and saline were administered orally on the 5th to 16th day in the corresponding 18 days groups,and the effects of blocking monocyte infiltration on neuralgia and anxiety-like behavior and the expressions of CD45 and 1BA-1 in CA3 region of hippocampus were observed.Results On the first day after SNI,the PWTs of mice in the 7 days and 14 days groups decreased and continued until before sacrifice(P＜0.01).The CD45 expression did little in the 7 days sham group.Compared with the sham group at the same time point,the CD45 expression did not increase in 7 days SNI mice((P＞0.05)and increased significantly in 14 days SNI mice(P＜0.01),only slightly co-expressed with IBA-1 and TMEM119 and no co-expression with GFAP and NeuN,the percentage of monocytes in the whole brain increased significantly in 14 days SNI mice(P＜0.01).Inhibition of microglial activation or CCR2 expression reduced the expression of CD45 in the CA3 in SNI mice(P＜0.01),increased the PWTs(P＜0.01)and alleviated anxiety-like behavior in SNI mice(P＜0.01).Conclusion There was an infiltration of peripheral monocytes in the hippocampal CA3 region after 14 days of SNI-induced neuralgia,which might be involved in the maintenance of neuralgia and the development of neuralgia-induced anxiety-like behaviors.

ObjectiveTo assess the predictive performance of four creatinine-based equations for estimated glomerular filtration rate （eGFR）： 2012 chronic kidney disease epidemiology collaboration （CKD-EPIcr） equation ， 2021CKD-EPIcr equation， Xiangya equation and European kidney function consortium （EKFC） equation. MethodsA total of 198 patients with chronic kidney disease from the Third Affiliated Hospital of Sun Yat-sen University and the Kiang Wu Hospital in Macau were enrolled. We compared the GFR measured （mGFR） by iohexol plasma clearance and the eGFR calculated by four equations. The agreement between mGFR and eGFR was analyzed by Bland-Altman plots， concordance correlation coefficient （CCC）， coverage probability （CP） and total deviation index （TDI）. The performance of eGFR equations， including their bias， precision， root square mean error （RSME）， and percentage of estimates within 30% deviation of measured GFR （P₃₀）， were evaluated. Bootstrap method （2 000 samples） was used to calculate bias， interquartile range （IQR）， RSME， and 95% confidence intervals （CI） for P_30. After selecting the optimal eGFR equation as the reference， we statisticlly tested other equations by ① Wilcoxon signed-rank test for bias； ② McNemar-Bowker test for P₃₀； ③ comparing RMSE and IQR with independent samples t test after 2 000 bootstrap samples were obtained. ResultsThe median mGFR and four eGFR equations （EKFC， 2012CKD-EPIcr， 2021CKD-EPIcr and Xiangya equation） in the overall population were 56.2 mL·min^-1·（1.73m²）^-1， 67.1 mL·min^-1·（1.73m²）^-1， 73.0 mL·min^-1·（1.73m²）^-1， 66.9 mL·min^-1·（1.73m²）^-1 and 63.8 mL·min^-1·（1.73m²）^-1， respectively. The Bland-Altman plots showed that EKFC equation had the lowest mean difference and the narrowest 95% limit of agreement. The EKFC equation had the optimal performance on CCC， TDI and CP with values of 0.90， 24.41 and 0.50， respectively. Overall， the bias， accuracy， P₃₀ and RSME from the EKFC equation was -0.99， 14.64， 0.80， and 14.68， respectively， with 95% CI ranging from -2.53 to 0.94， 11.82 to 17.35， 0.73 to 0.85， and 12.69 to 17.35， respectively， which were superior to those values from other three eGFR equations. The differences were statistically significant （all P < 0.05）. The results in the mGFR subgroups were basically consistent with the overall trend. ConclusionsOf the four eGFR equations validated in this study， the EKFC equation comprehensively surpasses 2012CKD-EPIcr equation， 2021CKD-EPIcr equation， and Xiangya equation. With P₃₀>75%， the EKFC equation can meet clinical diagnostic needs. Therefore， the EKFC equation is recommended for estimating GFR in a Chinese population， but more participants need be included to further support this conclusion.

ObjectiveTo investigate the predictive effect of high density lipoprotein （HDL） to C-reactive protein （CRP） ratio （HDL/CRP） on the progression of chronic kidney disease （CKD） in non-dialysis patients. MethodsNon-dialysis chronic kidney disease patients with at least two sets of follow-up data from the Third Affiliated Hospital of Sun Yat-sen University （Tian-he and Ling-nan districts）from 2015 to 2019 were enrolled. The baseline demographic characteristics and biochemical examination results were collected from the electronic medical record system. The patients were grouped according to the quantile of Ln（HDL/CRP）. The demographic and biochemical data were compared among groups by one-way ANOVA for normal distribution continuous variables， Kruskal-Wallis rank-sum test for non-normal distribution continuous variables， and Chi-square analysis for categorical variables. The relationship between HDL/CRP and baseline eGFR was investigated by correlation analysis， univariate and multivariate linear regression analysis. The Cox survival analysis were used to investigate the predictive effect of Ln（HDL/CRP） on renal deterioration events. ResultsTotally 9 142 patients with CKD were enrolled， and 439 patients were included in the end. There were 100 patients （22.8%） with chronic glomerulonephritis， 145 patients （33%） with diabetic nephropathy， 40 patients （9.1%） with hypertensive nephropathy， and 154 patients （35.1%） with other causes. According to Ln（HDL/CRP） quartile， group Quartile4 had a lower incidence of renal deterioration than the other three groups （11% vs. 21.1% to 21.8%） and had the highest baseline eGFR level. From Quartile1 to quartile 4 groups， age， Hba1c and APOA1 levels decreased gradually. The prevalence of chronic heart failure， BMI， hemoglobin， albumin， TC， LDL， TG， APOB100 levels were different among groups. Through correlation analysis， Ln （HDL/CRP） were positively correlated with baseline eGFR（r=0.162， P=0.001）. After adjusting for a variety of factors by Cox regression analysis， Ln （HDL/CRP） could be included in the final equation when defined deterioration of renal function as end point ［HR=0.79， 95%CI （0.69， 0.91）， P=0.001］. ConclusionHDL/CRP can reflect the severity of chronic kidney disease， and the ratio of HDL and CRP can predict the progression of chronic kidney disease in non-dialysis patient.

ObjectiveTo investigate the association of metabolic syndrome （MetS） and its components with rapid estimated glomerular filtration rate （eGFR） decline. MethodsThe China health and retirement longitudinal study （CHARLS） is a nationally representative cohort study focusing on physical and psychological health as well as social activities of middle-aged and elderly households. Cohort data could be obtained free of charge after application at the CHARLS website. We screened baseline data in 2011 and follow-up data in 2015 of CHARLS， selected those populations aged 45 and older， eGFR≥60 mL·min^-1·（1.73m²）^-1， without previous malignant tumors， heart disease， stroke and kidney disease， excluding those with missing values and outliers. Rapid eGFR decline was defined as an average annual decrease in eGFR of more than 3 mL·min^-1·（1.73m²）^-1. The 4400 participants enrolled were categorized into rapid eGFR decline group （770） and non-rapid eGFR decline group （3 630）. Logistic regression was performed to test the association of MetS and its components with rapid decline of eGFR. ResultsCompared with subjects without MetS， in subjects with MetS， we found no association between MetS and rapid eGFR decline after controlling for confounding factors （P>0.05）， with an OR （95%CI） of 0.988 （0.811~1.204）. After adjustment for confounding factors， the MetS components associated with rapid eGFR decline were central obesity （OR = 0.968， 95% CI： 0.778~1.204， P = 0.767）， elevated blood glucose （OR = 0.840， 95% CI： 0.715~0.986， P = 0.032）， hypertension （OR = 1.087， 95% CI： 0.919~1.285， P = 0.328）， decreased high-density lipoprotein cholesterol （OR = 1.085， 95% CI： 0.895~1.316， P = 0.404） and elevated triglyceride （OR = 1.110， 95% CI： 0.918~1.343， P = 0.281）. ConclusionsMetS was not an independent risk factor of rapid eGFR decline in the middle-aged and elderly populations， and elevated blood glucose was associated with the reduced risk of rapid eGFR decline. The false renal benefit caused by elevated blood sugar might be related to the renal hyperperfusion and hyperfiltration in the early stage of the disease. We need further follow-up to observe the dynamic effects of the MetS and its components on renal function.

BACKGROUND:The aim of this study is to investigate the diagnostic and prognostic value of neutrophil CD64 (nCD64) as a novel biomarker in sepsis patients. METHODS: One hundred fifty-one adult patients diagnosed with sepsis and 20 age-matched healthy controls were enrolled in the study. Patients with sepsis were further subdivided into a sepsis group and a septic shock group. nCD64 expression, serum procalcitonin (PCT) level, C-reactive protein (CRP) level, and white blood cell (WBC) count were obtained for each patient, and Sequential Organ Failure Assessment (SOFA) scores were calculated. RESULTS: nCD64 expression was higher in the sepsis group with confirmed infection than in the control group. The receiver operating characteristic (ROC) curve of nCD64 was higher than those of SOFA score, PCT, CRP and WBC for diagnosing infection. The area under the curve (AUC) of nCD64 combined with SOFA score was the highest for all parameters. The AUC of nCD64 for predicting 28-day mortality in sepsis was signifi cantly higher than those of PCT, CRP, and WBC, but slightly lower than that of SOFA score. The AUC of nCD64 or PCT combined with SOFA score was signifi cantly higher than that of any single parameter for predicting 28-day mortality. CONCLUSION: nCD64 expression and SOFA score are valuable parameters for early diagnosis of infection and prognostic evaluation of sepsis patients.

To investigate the preventive effect and possible mechanism of puerarin(Pur) in rat model of disuse osteoporosis(DOP),thirty healthy Wistar female rats of 2 months old were randomly divided into control group(Control), hindlimb suspension group(HLS), and puerarin group(HLS+Pur) in hindlimb suspension, with 10 rats in each group. A disuse osteoporosis model was established by tail suspension method, and 15.4 mg·kg~(-1) puerarin suspension was administered to HLS+Pur group every day, and the same volume of distilled water was administered to Control group and HLS group respectively. After 28 days, the rats were sacrificed by abdominal aorta blood collection, the main organs of the rats were removed, and the bone tissues of the rats were dissected. The organ index of the rats was calculated and the histopathology of the organs was observed under microscope. Bone mineral density test and bone biomechanical experiment were performed. Bone histomorphometry results were observed after bone tissue sectioning, and serum biochemical markers of bone metabolism were determined. There was no significant difference in organ index between the groups. There was no obvious abnormality in the pathological examination of the organs. The results of bone mineral density showed that puerarin could significantly increase the bone density of the tibia and vertebrae caused by hindlimb suspension. The mechanical parameters experiments showed that puerarin could effectively increase the maximum load and elastic modulus of the tibia and vertebrae. Fluorescence labeling showed that the fluorosis interval increased and the bone formation increased during puerarin treatment. The VG staining results showed that compared with the HLS group, in the puerarin group, the number of trabecular bone increased, the thickness of the trabecular bone became thicker, and the bone separation became smaller, which greatly improved the bone microstructure after hindlinb suspension. In addition, serum biochemical indicators showed that puerarin could promote bone formation index bone calcium. The content of osteocalcin(OC) increased and inhibited the formation of tartrate-resistant acid phosphatase 5 b(TRACP 5 b). Puerarin has a preventive effect in the rat model of disuse osteoporosis and its effect is good, and its mechanism may be related to promoting bone formation and inhibiting bone resorption.
Animals ; Bone Density ; Female ; Isoflavones ; pharmacology ; Osteocalcin ; metabolism ; Osteoporosis ; drug therapy ; Rats ; Rats, Wistar ; Tartrate-Resistant Acid Phosphatase ; metabolism

Objective To investigate the relationship between hemoglobin and blood pressure of pregnant women in Zhoushan islands, so as to provide scientific evidence for the etiological study of gestational hypertension. Methods A retrospective study was conducted among 1 383 pregnant women who received perinatal care in Zhoushan Maternal and Child Health Hospital of Zhejiang Province from January 2017 to June 2018. Pregnant women were monitored for hemoglobin content and blood pressure in the early, middle and late pregnancy. The multivariate linear regression was used to analyze the relationship between hemoglobin content and blood pressure in different pregnancy. Results The incidence of anemia in early, middle and late pregnancy was 7.74%, 25.45% and 15.76% respectively. The multivariate linear regression showed that hemoglobin levels during pregnancy had effects on systolic blood pressure in early, middle and late pregnancy, and the earlier hemoglobin levels were monitored, the more obvious the effect on systolic blood pressure was.With the increase of hemoglobin level, systolic blood pressure increased, such as the effect of hemoglobin on systolic blood pressure in early pregnancy, mid-pregnancy and late pregnancy. Hemoglobin of first trimster had the greatest effect (β=0.10, P<0.001), Hemoglobin of second trimester had no obvious effect, and that of third trimester had the second effect (β=0.04, P=0.027).Hemoglobin levels and diastolic blood pressure levels were similar to their relationship with systolic blood pressure. Conclusions Hemoglobin levels during pregnancy have significant effects on systolic and diastolic blood pressure in first, second and third trimsters of pregnancy. Regular measurement of hemoglobin levels during pregnancy can improve the health of pregnant women.

BACKGROUNDImmune cells within a tumor microenvironment have shown modulatory effects on tumor angiogenic activity. Renal cell carcinoma (RCC) is a hypervascular tumor that reportedly increases the frequency of regulatory T cells (Tregs) in tumor tissues. This study investigated the correlation between Tregs infiltration and angiogenic status in RCC.

METHODSThirty-six patients with RCC were enrolled in the present study, and twenty age-matched healthy donors were included as the control. Tregs were defined as CD4(+)CD25(high)CD127(low/-) T cells. The frequency of Tregs in peripheral blood and tumor infiltrating lymphocytes (TILs) were determined by flow cytometry. The expression of vascular endothelial growth factor (VEGF) in surgical resection specimens were measured with a commercial enzyme-linked immunosorbent assay (ELISA) kit. Microvessel density (MVD) was calculated on slides stained with CD34 antibody. Spearman's rank correlation was performed to evaluate the correlation between the frequencies of Tregs in TILs and VEGF values, as well as between frequencies of Tregs and MVD determinations.

RESULTSCompared to healthy controls, the frequency of peripheral blood Tregs was significantly increased in patients with RCC (P < 0.05). The percentage of tumor-infiltrating Tregs was higher than that of peripheral blood Tregs in patients with RCC (P < 0.01). In addition, the frequency of tumor-infiltrating Tregs was shown to significantly correlate with the pathological stage (P < 0.05) and nuclear grade (P < 0.01). Importantly, a significant positive correlation was observed between the frequency of tumor-infiltrating Tregs and VEGF protein expression (r = 0.51, P < 0.05), as well as between frequencies of Tregs and MVD score (r = 0.39, P < 0.05).

CONCLUSIONSThese observations suggest that the high pro-angiogenic status of RCC may be associated with the accumulation of Tregs in the local microenvironment. Angiogenesis networks may be connected with immune tolerance units and cooperate with each other to facilitate tumor growth and progression.

Adult ; Aged ; Carcinoma, Renal Cell ; immunology ; metabolism ; Cells, Cultured ; Enzyme-Linked Immunosorbent Assay ; Female ; Flow Cytometry ; Humans ; Immunohistochemistry ; Kidney Neoplasms ; immunology ; metabolism ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Neovascularization, Pathologic ; immunology ; metabolism ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo investigate the quantity and the pathway to damage hematopoietic cells of CD8+CD25+ and CD8+ HLA-DR+ effector T cells in peripheral blood (PB) of severe aplastic anemia(SAA) patients and explore the immunopathogenesis of SAA.

METHODSThe quantity of CD8+ CD25+ and CD8+ HLA-DR+ cells in PB and the expressions of perforin, granzyme B, tumor necrosis factor-beta (TNF-beta) and FasL in 29 SAA (14 untreated and 15 recovered) patients and 12 normal controls were analyzed by flow cytometry.

RESULTSThe fraction of CD8+ CD25+ T cells in CD8+ T cells was (3.67 +/- 2.58)% in untreated SAA patients, (5.19 +/- 4. 29)% in recovered patients and (4.84 +/- 2.31)% in normal controls, and that of CD8+ CD25+ T cells in CD3+ cells in the three groups was (2.25 +/- 1.35)%, (2.98 +/- 1.35)% and (2.11 +/- 1.88)%, respectively. They had no statistic difference among the 3 groups (P >0.05). The fraction of CD8+ HLA-DR+ T cells in CD8+ T cells was (39.30 +/- 8.13)% in untreated patients, which was significantly higher than that in recovered patients [(20.65 +/- 5.38)%] and controls [(18.34 +/- 6.68)%] (P<0.001), while there was no statistic difference between the latter two groups (P>0.05). CD8+ HLA-DR+ T cells in CD3+ cells was (27.81 +/- 7.10)% in untreated group, which was significantly higher than that of recovered group [(12.02 +/- 3.03)%] and controls [(8.50 +/-2.33)%] (P<0.01). And that in recovered group was higher than that in control group (P<0.05). The expressions of perforin, granzyme B, TNF-beta and FasL of CD8+ HLA-DR+ T cells in untreated group were 8.51%, 96.08%, 72.11% and 94.25% respectively, which were higher than those in recovered group (1.78%, 85.20%, 34.38% and 51.20%) and controls (1.86%, 82.09% ,17.92% and 32.91%). There was no statistic difference between recovered patients and controls (P>0.05).

CONCLUSIONThere were elevated quantity of CD8+ HLA-DR+ T cells and high expressions of perforin, granzyme B, TNF-beta and FasL in SAA, which might contribute to the bone marrow failure.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; metabolism ; pathology ; CD8-Positive T-Lymphocytes ; cytology ; Case-Control Studies ; Child ; Fas Ligand Protein ; metabolism ; Female ; Granzymes ; metabolism ; Humans ; Lymphocyte Count ; Lymphotoxin-alpha ; metabolism ; Male ; Middle Aged ; Perforin ; metabolism ; Young Adult

Objective To study the vibration frequencies and resonant peaks of different molecular groups in anti-diabetic drugs and to investigate the absorption spectra of these drugs in the range of terahertz, so as to accurately and efficiently identify similar drugs and provide foundation for understanding the contribution of such vibration in different molecular groups to pharmacology. Methods Using terahertz time domain spectroscopy (THz-TDS), six kinds of similar diabetes pills for treating diabetes mellitus (DM), including gliquidone, glipizide, gliclazide, glimepiride, repaglinide and metformin were tested to obtain their absorption spectra from 0.3~3.0 THz. Support vector machines (SVM) method was employed to separate these anti-diabetic drugs by selecting the experimental data from 1.5~2.0 THz. Results For gliquidone, glipizide, gliclazide, and glimepiride, an obvious resonance peak was found at 1.37 THz. According to the THz spectra, repaglinide and metformin could be easily separated from sulfonylurea drugs. Furthermore, with the help of SVM, the accuracy of discrimination for four kinds of sulfonylurea could reach 100%. Conclusions THz technology, as a new testing method, shall play a positive role in the drugs for their identification, quality control and distinguishing their chemical bonds/functional group.