BACKGROUND:It has been shown that Gushukang affects bone metabolism by regulating nucleotide and amino acid metabolism and immune mechanisms.Current research on the mechanism of Gushukang in the treatment of osteoporosis primarily focuses on osteoblast regulation and requires further improvement from the perspective of osteoclasts. OBJECTIVE:To investigate the mechanism by which Gushukang interferes with osteoclasts in the treatment of osteoporosis using RAW264.7 cells as the research model. METHODS:Twenty-four 8-week-old female Sprague-Dawley rats were randomly divided into four groups(n=6 per group):the three experimental groups were given 1,2 and 4 g/kg osteoporosis solution by gavage(2 times per day),and the control group was given an equal amount of distilled water by gavage(2 times per day).After 7 days of intragastric administration,aortic blood samples were extracted to collect serum samples using centrifugation,and serum samples from the same groups were combined to obtain the low-,medium-,and high-concentration Gushukang-containing and normal sera for the subsequent experiments.(1)RAW264.7 cells were cultured in six groups:normal serum was added to the control group;low,medium,and high concentration groups were added with low,medium,and high concentrations of Gushukang-containing serum,respectively;ML385,a nuclear factor erythroid 2-related factor 2(Nrf2)inhibitor was given in the Nrf2 inhibitor group;and t-BHQ,a Nrf2 activator,was added in the Nrf2 activator group.Cell viability was detected using the cell counting kit-8 assay.(2)The 3rd generation RAW 264.7 cells were cultured and divided into five groups:the blank control group was added with normal serum,the osteoclast group was added with receptor activator of nuclear factor κB ligand(RANKL),and the low-,medium-,and high-concentration groups were added with low-,medium-,and high-concentration Gushukang-containing serum based on the addition of RANKL.Tartrate-resistant acid phosphate staining was performed after 5 days of culture.(3)RAW264.7 cells were cultured and divided into five groups:blank control group was cultured with normal serum,osteoclast group cultured with normal serum and RANKL,high concentration+osteoclast group cultured with RANKL+high concentration Gushukang-containing serum,osteoclast+Nrf2 agonist group cultured with RANKL+t-BHQ,and high concentration+osteoclast+Nrf2 inhibitor group cultured with RANKL+high concentration Gushukang-containing serum+ML385.Western blot assay and determination of reactive oxygen content were performed after 5 days of culture. RESULTS AND CONCLUSION:The cell counting kit-8 results indicated that Gushukang-containing serum,NRF2 inhibitor or agonist had no significant effect on RAW264.7 cell viability.Tartrate-resistant acid phosphate staining results demonstrated that Gushukang-containing serum exhibited a concentration-dependent inhibitory effect on osteoclast differentiation.Western blot analysis and determination of reactive oxygen species revealed that compared with the blank control group,Nrf2 protein expression was decreased in the osteoclast group(P<0.05),while c-Fos and NFATc1 protein expression and reactive oxygen species content were elevated(P<0.05);compared with the osteoclast group,Nrf2 protein expression was elevated and reactive oxygen species content was decreased in the high-concentration+osteoclast group,osteoclast+Nrf2 agonist group,and high-concentration+osteoclast+Nrf2 inhibitor group(P<0.05),while c-Fos and NFATc1 protein expression was decreased in the high concentration+osteoclast group and osteoclast+Nrf2 agonist group(P<0.05);compared with the high concentration+osteoclast group,Nrf2 protein expression was decreased(P<0.05)and reactive oxygen species content was elevated(P<0.05)in the high concentration+osteoclast+Nrf2 inhibitor group.To conclude,Gushukang reduces reactive oxygen species production by activating Nrf2,thereby inhibiting downstream of the c-Fos/NFATc1 pathway and suppressing osteoclast differentiation.

ObjectiveTo explore the amelioration of cognitive dysfunction in diabetes mellitus （DM） by Jianpi Qinghua prescription （JPQH） based on type 2 diabetes （T2DM） model rats. MethodFifty healthy male Wistar rats of SPF grade were randomly divided into control group （n=10） and experimental group （n=40）. The rats in the control group were fed conventionally， while those in the experimental group were fed on a high-sugar， high-fat diet for six weeks and administered with streptozotocin （STZ） for the induction of the DM model. The model rats were randomly divided into model group， sitagliptin group （1.2 g·L^-1）， pioglitazone group （0.8 g·L^-1）， and JPQH group （1.3 g·mL^-1）， with 10 rats in each group. After six weeks of drug intervention， the changes in body weight， blood glucose， and other related indexes of each group were recorded. Enzyme-linked immunosorbent assay （ELISA） was performed to detect the levels of tumor necrosis factor-α （TNF-α）， interleukin-1β （IL-1β）， and interleukin-6 （IL-6） in the peripheral blood and brain. The Morris water maze test was used to evaluate the cognitive function in rats. Hematoxylin-eosin （HE） staining was used to observe the pathological morphology of the hippocampal CA region. The amyloid β-protein 40 （Aβ₄₀） level was detected by immunohistochemistry. The protein expression of t-tau and p-tau in hippocampal neurons of rats was detected by Western blot. ResultCompared with blank group， the body weight of model group was significantly decreased （P<0.05）， blood glucose level was significantly increased （P<0.01）， inflammatory cytokines TNF-α and IL-1β were increased （P<0.05）， learning and spatial ability were significantly decreased （P<0.01）， the arrangement of hippocampal cells was loose and disordered， and the intercellular space was significantly increased. The number of cells decreased significantly， and the expression of Aβ₄₀ increased significantly. and increased t-tau and p-tau protein content in the hippocampus （P<0.01）. Compared with model group， the JPQH group showed reduced blood glucose （P<0.01）， decreased TNF-α and IL-1β levels in the peripheral blood and cerebrospinal fluid （P<0.05）， a downward trend of IL-6 without a statistical difference， improved learning and spatial memory ability （P<0.01）， densely arranged cells in the hippocampal CA1 area， increased cell number， reduced Aβ₄₀ expression， and decreased p-tau protein expression （P<0.05）. ConclusionJPQH can prevent cognitive dysfunction in DM by reducing inflammatory factor levels， decreasing neurotoxicity caused by Aβ₄₀ deposition， and inhibiting hyperphosphorylation of tau protein in DM rats.

Objective:To explore the anti-aging effects of berberine hydrochloride and underlying mechanism.Methods:Twelve 4-week-old C57BL6/J mice were divided into aging group (fed with normal chaw), aging+ BBR intervention group(fed with normal chaw containing 1 g/kg berberine hydrochloride). At the age of 64 weeks, all the experimental mice were executed. The liver tissues were made into paraffin sections for HE staining to observe the morphological and structural integrity of liver parenchymal cells for pathological evaluation. Immunofluorescence was used to detect p16 protein expression levels in liver. The expression levels of malondialdehyde(MDA), superoxide dismutase(SOD), and glutathione peroxidase(GSH-Px) were detected by colorimetry. The expression levels of interleukin(IL)-1β, IL-6, and tumor necrosis factor-α(TNF-α) in liver tissues and IL-8 in serum were detected by enzyme linked immunosorbent assay(ELISA) technique. The p16, p21, nuclear factor erythroid-2 related factor 2(Nrf2), nuclear factor-κB(NF-κB), phospho(p-)NF-κB, inhibitory κB(IκB)α, p-IκBα, and p38 mitogen-activated protein kinase(MAPK) protein expression levels in liver were detected by Western blotting. The same indices were tested in the 16-week-old mice as the young control group.Results:Compared with the young control group, the liver tissue in the aging control group exhibited morphological aging and antioxidant capacity were reduced( P<0.01), and inflammatory factors were increased( P<0.05 or P<0.01). Compared with the aging group, the liver tissues in the aging+ BBR intervention group were still maintain regular arrangement of hepatocytes, the p16 and p21 protein expression levels were significantly increased( P<0.05 or P<0.01), the antioxidant capacity were increased( P<0.05 or P<0.01), the level of inflammatory factors were decresed( P<0.05 or P<0.01), and the p38 MAPK/NF-κB pathway and Nrf2 pathway were inhibited( P<0.001). Conclusion:Berberine hydrochloride improves the aging morphology of the liver, potentially by suppressing the p38 MAPK/NF-κB pathway to reduce inflammation levels and inhibiting the Nrf2 pathway to ameliorate oxidative stress.

Objective:To investigate the hepatitis B surface antigen (HBsAg) clearance condition and its predictive factors after treatment with nucleos(t)ide analogues to pegylated interferon-α add-on therapy in patients with chronic hepatitis B.Methods:Patients with chronic hepatitis B who visited the First Affiliated Hospital of Zhengzhou University from 2018~2019 were prospectively enrolled. HBsAg≤ 1500 IU/mL, hepatitis B e antigen-negative, HBV DNA undetectable, received antiviral treatment with nucleos(t)ide analogues for at least one year, and pegylated interferon-α add-on therapy for 48 weeks were included. The primary endpoint of study was to determine the proportion of HBsAg clearance at 72 weeks. Concurrently, the predictive factors for HBsAg clearance were analyzed. Quantitative and qualitative data were analyzed using a t-test or non-parametric test and a Fisher's exact test.Results:A total of 38 cases were included in this study, of which 13 cases obtained HBsAg clearance at 48 weeks of therapy and another six cases obtained HBsAg clearance throughout the extended treatment period of 72 weeks, accounting for 50.00% of all enrolled patients. There was a significant difference in HBsAg dynamics between the HBsAg clearance group and the non-clearance group (P < 0.05). Univariate logistic regression analysis showed that patients' age, baseline, 12-and 24-week HBsAg levels, and early HBsAg reduction were predictive factors for HBsAg clearance at 72 weeks of treatment. Multivariate logistic regression analysis showed that age (OR = 1.311; P = 0.016; 95% confidence interval: 1.051~1.635) and HBsAg levels at 24 weeks of treatment (OR = 4.481; P = 0.004; 95% confidence interval: 1.634~12.290) were independent predictors for HBsAg clearance.Conclusion:Hepatitis B e antigen-negative, nucleos(t)ide analogue treated, HBsAg ≤ 1500 IU/mL, and HBV DNA undetectable, peg-IFNα add-on treatment for 48 weeks could promote HBsAg clearance in patients with chronic hepatitis B. Six of the sixteen cases (37.50%) who did not obtain HBsAg clearance at week 48 did so with the course of therapy extended to week 72. Hence, the optimal individualized treatment strategy should be customized according to the predictors rather than the fixed 48-week course. Age (≤ 38), baseline HBsAg level (≤2.86 log 10IU/ml), HBsAg level at 24 weeks (≤ 0.92 log 10IU/ml), and 12-week HBsAg decrease from baseline (≥ 0.67 log 10IU/ml) indicate that patients are highly likely to obtain HBsAg clearance at the 72 weeks of combination therapy, in which the combined indicator based on HBsAg level ≤0.92 log 10IU/ml at 24 weeks will identify 85.0% to 100.0% of patients with HBsAg clearance.

Objective:To evaluate the effect of Parkinson′s disease factor on the sedative efficacy of dexmedetomidine.Methods:The patients of either sex, aged 45-64 yr, of American Society of Anesthesiologists Physical Status classification Ⅱor Ⅲ, with body mass index of 18.5-30.0 kg/m 2, undergoing non-intracranial space-occupying lesions in neurosurgery, were selected.Patients were divided into control group (group C) and Parkinson′s disease group (group P) according to whether they had Parkinson′s disease or not.The ED 50 of dexmedetomidine was determined by using the Dixon′s up-and-down method.The initial dose of dexmedetomidine was 0.5 μg/kg in both groups, and each time the concentration increased/decreased by 0.05 μg/kg in the next patient, which was repeated until 7th independent crossover pair (loss of consciousness) appeared, and then the test was ended.The ED 50 and 95% confidence interval of dexmedetomidine inducing loss of consciousness were calculated using the probit test in a Logistic regression model.Hypertension, hypotension, bradycardia and nausea and vomiting were recorded. Results:Compared with group C, the ED 50 of dexmedetomidine inducing loss of consciousness was significantly increased in group P ( P<0.05), and no significant change was found in the incidence of adverse reactions in group P ( P>0.05). Conclusions:Parkinson′s disease factor can decrease the sedative efficacy of dexmedetomidine.

Objective To investigate the diagnostic value of the texture analysis in differentiating adult pilocytic astrocytomas (PA)from hemangioblastomas(HB).Methods 22 adult patients with PA and 20 patients with HB which were confirmed by postoperative pathological were retrospectively reviewed.The conventional MRI features and texture parameters were analyzed.Eight texture parameters were extracted using run-length matrix(RLM),and the differences of texture parameters of the two groups were analyzed by independent-samples t test.Results The short run emphasis(SRE),grey-level non-uniformity(GLNU),run-length non-uniformity(RLNU),high grey-level run emphasis(HGRE)and short run high grey-level emphasis(SRHGE)were higher in adult PA than in HB.The long run emphasis(LRE),low grey-level run emphasis(LGRE)and short run low grey-level emphasis(SRLGE)were lower in adult PA than in HB.The eight texture parameters had significant differences between the two groups(P<0.05).Conclusion Texture analysis can provide reliably objective basis for differentiating adult PA from HB.

A total of 175 patients with type 2 diabetes mellitus (T2DM) were composed of 69 with normoalbuminuria,60 with microalbuminuria,and 46 with macroalbuminuria.The control group consisted of 64 healthy individuals.Serum YKL-40 levels were determined with ELISA method and related metabolic data were collected.Serum YKL-40 levels were significantly higher in T2DM group than in control group(P＜0.01).Significant correlations of YKL-40 were found with the ratio of microalbuminuria to uric creatinine(r=0.677,P＜0.01),HbA1C (r =0.562,P＜0.01),systolic blood pressure (r =0.372,P =0.001),HOMA-IR (r =0.460,P =0.001),low density lipoprotein-cholesterol (r=0.304,P=0.012),age(r=0.260,P=0.015),blood uric acid (r=0.329,P=0.018),and high density lipoprotein-cholesterol (r=-0.247,P=0.032).YKL-40 may play a role in the occurrence and development of diabetic nephropathy.

Periostin is a matri-cellular protein which was originally identified in MC3T3-E 1 osteoblast-like cell line,expressing in multiple tissues like bones,teeth,skin and cardiac valves.Periostin is also found in a large variety of cancers and injured tissues,involving in cancer cell invasion and metastasis as well as wound repair.Recent studies have suggested the role of Periostin in osteoblast adhesion and differentiation,fibrillogenesis,mineralization and bone fracture healing,and its expression is regulated by mechanical stress,various transcription factors,hormones and growth factors.In this article,we will discuss the expression,localization and general characteristics of Periostin,and provide a review on the study of it in bone biology.

Objective To investigate the neonatal death causes and the relationship between the causes and the neonatal age,fetus age and the year in order to reduce neonatal mortality.Methods Data of neonatal autopsy and pathological records from 107 neonates in our hospital from January 1998 to May 2008 were collected,analyzed and summarized.Results 40.2%of the cases died within 24 hours and 72.9%died within 7 days after birth.The main death cause for those who died within 7 days was asphyxia(48.6%),and the main death causes ior those who died after 7 days were deformity(34.5%)and hemorrhage(34.5%).The most common death cause for premature bahies Was hyaline membrane disease(29.7%),and the most common one for term infants was congenital heart disease(23.3%).In the past 5 years.asphyxia Was still the most common death causein our country,those who died of infection were on the rise and the deformed babies increased too;on the contrary.those who died of obstetric causes and hemorrhage decreased.Conclusion Neonatal mortality could be reduced by prenatal management,prevention of premature delivery,improvement of the cooperation between pediatrics and obstetrics,and better management of NICU.