Eosinophilic vacuolated tumor of the kidney: clinicopathological and molecular characteristics / 中华病理学杂志

Objective: To study the clinicopathological features, immunophenotype, molecular changes, differential diagnosis and prognosis of eosinophilic vacuolated tumor (EVT) of the kidney. Methods: Four cases were collected retrospectively from 2014 to 2020 at Ningbo Diagnostic Pathology Center. The clinicopathologic features and immunophenotypic profile were studied by light microscopy and immunohistochemistry. Targeted next-generation sequencing (NGS) panel was used to detect cancer-associated mutation. Follow-up and literature review were also performed. Results: Among the 4 patients studied,2 were males and 2 were females. The age of the patients ranged from 44 to 63 years (the mean age: 51 years).Tumor size ranged from 1.5 to 4.2 cm (mean: 2.3 cm). Microscopically, tumors were well-circumscribed, unencapsulated. Thick-walled vessels and entrapped renal tubules were found within or at the periphery of the tumors. The tumors were predominantly composed of nest pattern, and focal tubular pattern. The tumor cells exhibited abundant, eosinophilic, granular cytoplasm and conspicuous, large nucleoli. Prominent intracytoplasmic vacuoles were seen. These cytoplasmic vacuoles varied in size and frequently coalesced into a large space. Loose fibromatous or hyaline stroma was focally noted. Immunohistochemically, the tumor cells in all cases exhibited a CD117+/CK7-phenotype. All cases were positive for CD10 and p504s. MTOR, S6 and cathepsin K were positive in 4 cases. TFE3, CA9, Melan A and HMB45 were negative in all cases. SDHB retained expression. NGS demonstrated MTOR mutations in all cases, and TSC2 mutation in 2 cases. Conclusions: EVT is a rarely oncocytic renal tumor with unique morphology, immunohistochemical phenotype, molecular profile and an indolent behavior. Recognition of the characteristics of this novel but rare entity will allow for better classification of renal tumors.

Anaplastic lymphoma kinase-translocation renal cell carcinoma: clinical and pathological analysis / 中华病理学杂志

Objective: To investigate the clinicopathological features, molecular characteristics, differential diagnosis and prognosis of anaplastic lymphoma kinase (ALK)-translocation renal cell carcinoma. Methods: Two cases of ALK-translocation renal cell carcinoma diagnosed from January 2011 to December 2020 were retrospectively analyzed to characterize their morphological features, immunohistochemical expression and prognosis. Multiple molecular studies including fluorescence in situ hybridization (FISH), reverse transcriptase-polymerase chain reaction (RT-PCR), and next-generation sequencing were performed to characterize the genetic alterations. Results: Two patients included one male and one female, with 59 and 57 years old, respectively. Morphologically, case 1 resembled collecting duct carcinoma or renal medullary carcinoma, which demonstrated tubular, microcapsule and reticular structures, with a remarkable myxoid background and lymphocytes infiltration; case 2 resembled Xp11.2 translocation renal cell carcinoma or type 2 papillary renal cell carcinoma, which demonstrated tubular papillary and focal solid structures, with flocculent cytoplasm and many foamy histiocytes, but without myxoid background and lymphocytes infiltration. Immunohistochemistry showed strongly positive expression of ALK. CK7, E-cadherin, vimentin, PAX8 and CD10 showed various degrees of expression, and other antibodies were nonreactive. A variety of molecular assays showed definite ALK gene translocation, with rare VCL-ALK gene fusion (VCL exon and 16-ALK exon 20) in case 1, and EML4-ALK gene fusion (EML4 exon and 2-ALK exon 20) in case 2. Conclusions: ALK-translocation renal cell carcinoma is rare with various morphological features, and is easy to miss and misdiagnose. The characteristic ALK expression and molecular detection of ALK translocation are helpful for diagnosing this type of renal cell carcinoma.

Papillary renal neoplasm with reverse polarity: a clinicopathological analysis / 中华病理学杂志

Objective: To study the clinical pathological characteristics, immunophenotype, molecular changes and prognosis of the papillary renal neoplasm with reverse polarity (PRNRP). Methods: Nine cases of PRNRP, diagnosed from 2013 to 2019, were retrieved from the Department of Pathology of Nanjing Jinling Hospital, Nanjing University School of Medicine. Histomorphology, immunophenotype and molecular genetics were analyzed with review of the literatures. Results: There were five male and four female patients, aged from 49 to 70 years, with an average age of 60.1 years. During a mean follow-up of 29 months, one patient died for other cause, and the others survived without disease. Microscopically, the tumor cells arranged in papillary structure with a fibrovascular core, the surface of which was covered with a single layer of cuboidal or columnar cells. The most prominent feature was that the tumor nuclei located at the top of the cytoplasm far from the basement membrane, and they were monotonous in size and arranged neatly with no or few nucleoli. Immunohistochemically, all nine cases of PRNRP showed diffuse positive expression of CK7 and E-cadherin, various degrees of P504s expression, and no expression of CD10 and CD117, with a Ki-67 index of 1%-3%. Unlike other papillary renal cell carcinoma, the nine cases of PRNRP all showed characteristic positive expression of GATA3. The fluorescence in situ hybridization assay showed that the majority of PRNRPs (8/9) did not have triploids on chromosomes 7 and 17. The sequencing of the KRAS gene confirmed the presence of a nonsense KRAS mutation in 8 of the 9 cases. Conclusions: PRNRP is a subtype of papillary renal cell carcinoma with characteristic morphological, immunophenotypic and molecular features, and indolent behaviors. More data are needed to define PRNRP as "carcinoma", and a definitive diagnosis of PRNRP is of great significance for proper treatment choice and accurate prognostication.

Succinate dehydrogenase-deficient renal cell carcinoma:a clinicopathological, ultrastructural and molecular analysis / 中华病理学杂志

Objective: To investigate the clinicopathological features, immunophenotype, ultrastructure, genetic alterations and prognosis of succinate dehydrogenase-deficient renal cell carcinoma (SDH RCC). Methods: A total of 11 SDH RCCs, diagnosed from 2010 to 2019, were selected from the Department of Pathology of Nanjing Jingling Hospital, Nanjing University School of Medicine for clinicopathologic, immunohistochemical (IHC), ultrastructural investigation and follow-up. The molecular features of seven cases were analyzed by the panel-targeted DNA next generation sequencing (NGS). Results: There were seven males and four females, with ages ranging from 24 to 62 years (mean 41.4 years, median 41 years). Microscopically, SDH RCC was mainly composed of solid and tubular structures with local cystic change. Four cases showed nested or trabecular structure distributed in a loose hypocellular connective tissue or around scar, similar to oncocytoma. The neoplastic cells demonstrated flocculent eosinophilic cytoplasm with typical intracytoplasmic vacuoles. Immunohistochemically, eight cases were negative for SDHB; three cases showed focal and weak expression, whereas normal renal tubular and vascular endothelial cells demonstrated strong cytoplasmic staining. NGS of DNA targeted-panel detected pathogenic mutations of SDHB gene in seven cases (including three cases with equivocal IHC expression of SDHB), without any mutations in other SDH related genes. There were four cases of SDHB missense mutation, one case of frameshift mutation, one case of splicing mutation, and one case of acquired stop codon mutation. Conclusions: SDH RCC is a distinct variant of RCCs with genetic tendency or with hereditary cancer syndrome. NGS is recommended to detect the related gene mutations for a definitive diagnosis. The patients should be closely followed up.

EB virus-positive discordant lymphoma: report of 1 case and review of literature / 白血病·淋巴瘤

Objective:To investigate the clinical manifestations, laboratory tests, diagnosis and treatment of discordant lymphoma (DL).Methods:The clinical data of a patient with EB virus-positive DL admitted to Taizhou People's Hospital in November 2019 were retrospectively analyzed and the related literature was reviewed.Results:The patient underwent a cervical lymph node biopsy pathology examination at onset, and then results suggested angioimmunoblastic T-cell lymphoma (AITL). The patient subsequently developed gastrointestinal bleeding and underwent resection of small bowel lesions, and postoperative pathology suggested diffuse large B-cell lymphoma (DLBCL). The patient was finally diagnosed as DL. The R2-CHOP chemotherapy regimen was given to the patient, but the patient still had recurrent gastrointestinal bleeding and poor general condition. The patient refused chemotherapy and was changed to lenalidomide monotherapy. Finally, the patient died due to multiorgan failure, with an overall survival of 13 months.Conclusions:DL is rarely seen in lymphoma, whereas the combination of AITL and DLBCL is extremely rare. The clinicians need to improve the understanding of this disease to avoid misdiagnosis and missed diagnosis.

Influence of the number of high-risk cytogenetic abnormalities on the clinical characteristics and prognosis in 360 patients with newly diagnosed multiple myeloma / 中华血液学杂志

Objective: To investigate the influence of the number of high-risk cytogenetic abnormalities (HRCA) on the clinical characteristics and prognosis of patients with newly diagnosed multiple myeloma (MM) . Methods: A total of 360 patients with newly diagnosed MM admitted to Jiangsu Province Hospital between November 2013 and September 2020 were included in this study. Cytoplasmic light chain immunofluorescence with fluorescence in situ hybridization (cIg-FISH) was used to detect HRCA. Cytogenetic abnormalities were combined with clinical characteristics and outcomes for further analysis. Results: Among the 360 patients, 120 patients (33.3%) presented with no HRCAs, and 175 (48.6%) , 61 (16.9%) , and four (1.1%) patients had one, two, and three HRCA (s) , respectively. Patients were divided into three groups, including the no-HRCA group, one-HRCA group, and ≥two-HRCA group, according to the number of HRCAs. There were significant differences in the R-ISS stage, hemoglobin level, albumin level, and the proportion of bone marrow plasma cells among the three groups (P<0.05) . The COX proportional-hazards model identified extramedullary disease (P=0.018) , HRCA ≥ 2 (P=0.001) , and absence of autologous hematopoietic stem cell transplantation (P<0.001) as independent risk factors for progression free survival (PFS) and identified lactate dehydrogenase (LDH) level ≥ 220 U/L (P<0.001) , HRCA ≥2 (P=0.001) , and absence of autologous hematopoietic stem cell transplantation (P=0.005) as independent risk factors for overall survival (OS) . The median PFS was 28 months, 22 months, and 14 months (P=0.005) for the three cohorts, and their OS was not reached,60 months, and 30 months (P=0.001) , respectively. Conclusions: HRCA ≥ 2 is an independent risk factor for decreased survival in patients with newly diagnosed MM. More HRCAs result in heavier tumor burden, as well as a higher risk of disease progression and death.

Predicting the grades of Astragali radix using mass spectrometry-based metabolomics and machine learning / 药物分析学报

Astragali radix(AR,the dried root of Astragalus)is a popular herbal remedy in both China and the United States.The commercially available AR is commonly classified into premium graded(PG)and ungraded(UG)ones only according to the appearance.To uncover novel sensitive and specific markers for AR grading,we took the integrated mass spectrometry-based untargeted and targeted metabolomics ap-proaches to characterize chemical features of PG and UG samples in a discovery set(n=16 batches).A series of five differential compounds were screened out by univariate statistical analysis,including arginine,calycosin,ononin,formononetin,and astragaloside Ⅳ,most of which were observed to be accumulated in PG samples except for astragaloside Ⅳ.Then,we performed machine learning on the quantification data of five compounds and constructed a logistic regression prediction model.Finally,the external validation in an independent validation set of AR(n=20 batches)verified that the five com-pounds,as well as the model,had strong capability to distinguish the two grades of AR,with the pre-diction accuracy＞90％.Our findings present a panel of meaningful candidate markers that would significantly catalyze the innovation in AR grading.

Effect Mechanism of Total Flavonoid of Mori Cortex Combined with Total Saponins of Anemarrhenar Rhizoma on Hyperlipidemia Rats with Osteoporosis / 中国实验方剂学杂志

Objective:To explore the improvement effect of total flavonoids of Mori Cortex combined with total saponins of Anemarrhena Asphodeloide on hyperlipidemia rats with osteoporosis and its possible mechanism. Method:The 40 SPF male SD rats were adaptively fed for 7 days， and then randomly divided into normal group， model group， calcitriol group （45 ng·kg^-1）， total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloide 1∶2 group （0.6 g·kg^-1+0.4 g·kg^-1） and 2∶1 group （1.2 g·kg^-1+0.2 g·kg^-1）. Except for the normal group， rats in the other groups were fed with high fat for 9 weeks， the normal group and the model grouotal flavonoids of total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloip were given normal saline by gavage， and the other groups were given corresponding drugs by gavage， after 12 weeks of administration， except for the normal group ， the other groups were given intramuscular injection of glucocorticoids at the same time. After 22 weeks of administration， the weight of rats with total flavonoids from Mori Cortex combined with total saponins of Anemarrhena Asphodeloide was measured. Serum levels of total cholesterol （TC）， triglyceride （TG）， low density lipoprotein cholesterol （LDL-C）， osteocalcin （BGP） and bone alkaline phosphatase （BALP） were determined by biochemical assay. Hematoxylin-eosin （HE） staining to observe the pathological changes of rat tibia. Real-time fluorescent quantitative polymerase chain reaction （Real-time PCR） was used to detect the expression levels of peroxisomal proliferators activate the receptor gamma（PPAR<italic>γ</italic>） and Runt-related transcription factor 2 （Runx2） mRNA in rat bone tissue， immunofluorescence was used to detect the expression of PPAR<italic>γ</italic> and Runx2 in rats. Result:Compared with normal group， the body mass of rats in model group was significantly increased （<italic>P</italic><0.01）， and the contents of TC， TG， and LDL-C in the serum were significantly increased （<italic>P</italic><0.01）. Compared with model group， the body weight of rats in thet total flavonoids of Mori Cortex and total saponins of Anemarrhena Asphodeloide 1∶2 group and 2∶1 group were significantly reduced （<italic>P</italic><0.01）， and the contents of TC， TG， and LDL-C in the serum were significantly reduced （<italic>P</italic><0.01）， the content of BGP and BALP increased （<italic>P</italic><0.01）. HE staining results showed that compared with the normal group， the tibia fat vacuoles of the model group increased， and the number of osteoblasts decreased， compared with the model group， the total flavonoids of the Mori Cortex and the flavonoids-total saponins of Anemarrhena Asphodeloide 1∶2 group and 2∶1 group decreased in tibia fat vacuoles and increased the number of osteoblasts， the results of immunofluorescence and Real-time PCR showed that， compared with normal group， the expression of Runx2 in the model group decreased and the expression of PPAR<italic>γ</italic> increased （<italic>P</italic><0.01）. Compared with model group， the total flavonoids of Mori Cortex-total saponins 1∶2 group and the total flavonoids of Mori Cortex-total saponins 2∶1 Group up-regulated the expression of Runx2 and down-regulated the expression of PPAR<italic>γ </italic>（<italic>P</italic><0.05，<italic>P</italic><0.01）. Conclusion:The total flavonoids of Mori Cortex combined with the total saponins of Anemarrhena Asphodeloide up-regulated Runx2 and down-regulated the expression of PPAR<italic>γ</italic> mRNA and protein， thereby affecting the metabolism of TG and TC in the blood， achieving a therapeutic effect on osteoporosis， provides experimental basis for the clinical prevention and treatment of hyperlipidemia with osteoporosis.

Effect of peroxisome proliferator-activated receptor-γ coactivator-1α on liver injury induced by periodontitis in rats / 华西口腔医学杂志

OBJECTIVES@#To investigate the effect of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) on liver injury induced by periodontitis in rats.@*METHODS@#Twenty-four male Wistar rats were randomly divided into two groups: control group and periodontitis group, twelve per group. In periodontitis group, the periodontitis models were established for the maxillary first molars in rats by means of "wire ligation+vaccinationwith @*RESULTS@#The probing depth, tooth mobility and sulcus bleeding index in periodontitis group were significantly higher than that in control group. HE staining showed in periodontitis group, hepatic cords ranged disorderly and there were vacuoles in cells and inflammatory cells infiltrated in liver tissues of rats, and there was no obvious abnormality in control group. The qRT-PCR results showed that the mRNA expression levels of @*CONCLUSIONS@#PGC-1α may be involved in the process of periodontitis-induced liver injury in rats.

Effect of Anemarrhena Asphodeloside BⅡ on Osteoclast Differentiation Based on Molecular Docking / 中国实验方剂学杂志

Objective:To explore the effect of anemarrhena asphodeloside BⅡ (TBⅡ) on the expressions of nuclear transcription factor-κB receptor activator factor ligand （RANKL）, RANK and C-FOS genes during osteoclast differentiation. Method:Molecular docking software LeDock was used to score the docking of TBⅡ with RANKL, RANK and C-FOS. RAW264.7 was treated with soluble RANKL（sRANKL） and divided into control group, sRANKL group (model group), Icariin (Ica) group, low-dose TBIⅡ group (2 μmol·L-1), medium-dose TBⅡ group (4 μmol·L-1), and high-dose TBⅡ group (8 μmol·L-1). The corresponding kit was used to detect iconic enzyme (TRAP) of osteoclast differentiation. Total RNA was extracted by trizol method, Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR) was used to detect the expressions of C-FOS, upstream RANKL/RANK and downstream nuclear factor of activated T-cells cytoplasmic 1 (NFATC1), and osteoprotegerin OPG. Result:The molecular docking score were -11.86, -11.38, -12.34 kcal·mol-1, and there might be multiple binding sites between TBII as well as RANKL, RANK and C-FOS. Compared with the control group, the content of TRAP in model group increased significantly (P<0.01), and compared with model group, the content of TRAP in each administration group decreased significantly (P<0.01), and TBⅡ decreased the content of TRAP in a dose-dependent manner. Compared with the control group, the expressions of RANKL, RANK, C-FOS and NFATC1 increased (P<0.01), whereas the expression of OPG decreased (P<0.01) in model group. Compared with model group, the expressions of RANKL, RANK, C-FOS and NFATC1 decreased (P<0.01), while the expression of OPG increased (P<0.01) in each administration group. Conclusion:TBⅡ may inhibit the differentiation of osteoclast precursors into osteoclasts, inhibit osteoclast activity, reduce bone resorption and improve osteoporosis by regulating RANKL/RANK/C-FOS signal pathway.

Analysis on changes of fluoride in rural centralized water supply in Inner Mongolia Autonomous Region from 2014 to 2018 / 中华地方病学杂志

Objective:To understand the situation and distribution of fluoride in rural centralized water supply in Inner Mongolia Autonomous Region (Inner Mongolia for short), and provide a reference for adjustment of prevention and control measures.Methods:From 2014 to 2018, 836, 947, 1 033, 1 068, 1 099 rural centralized water supply monitoring sites were designated in 77 banners (counties, districts) separately in Inner Mongolia, and factory water and tip water samples were collected during plentiful phase and exhausted phase every year, respectively, and fluoride content was tested. Descriptive analysis was done according to time, water period (plentiful phase and exhausted phase), water sample type (factory water and tip water), water treatment method (conventional treatment, sedimentation and filtration, only disinfection and untreated), area distribution [eastern region (Hulunbuir, Xing'an League, Tongliao and Chifeng), central region (Hohhot, Baotou, Ulanqab and Xilinhot) and western region (Ordos, Bayannur and Alashan)], and the results were analyzed visually by ArcMAP 10.2.Results:From 2014 to 2018, 3 251, 3 671, 4 058, 4 087 and 4 395 water samples were collected, the medians fluoride were 0.69, 0.70, 0.69, 0.64 and 0.66 mg/L, and the annual compliance rates of fluoride were 80.31% (2 611/3 251), 81.83% (3 004/3 671), 83.14% (3 374/4 058), 85.91% (3 511/4 087) and 84.57% (3 717/4 395). The difference of compliance rate of fluoride in rural centralized water supply in different years was statistically significant (χ 2=51.748, P < 0.01), and the compliance rate of fluoride showed an increasing trend with the years (χ 2=41.140, P < 0.01). The compliance rates of fluoride in plentiful phase and exhausted phase were 83.36% (8 128/9 750) and 83.29% (8 089/9 712), respectively, and the difference was not significant statistically (χ 2=0.020, P > 0.05). As for water sample type, the compliance rates of fluoride in factory water and tip water were 83.55% (6 583/7 879) and 83.17% (9 628/11 576), and the difference was not significant statistically (χ 2=0.485, P > 0.05). The difference of compliance rate of fluoride in different water treatment methods was statistically significant (χ 2=192.014, P < 0.01). The compliance rates of fluoride in water with conventional treatment and only disinfection were higher [95.51% (404/423) and 94.44% (986/1 044)]; and the untreated water had the lowest compliance rate of fluoride [81.75% (13 073/15 991)]. There was a statistically significant difference in compliance rates of fluoride in the eastern, central and western regions (χ 2=629.256, P < 0.01), with the eastern region had the highest compliance rate of 89.17% (7 337/8 228); the central region had the lowest compliance rate of 74.67% (5 391/7 220). The visualization results showed that the compliance rate of fluoride was obviously low in the central region north of Yin Mountains and west of Greater Higgnan Mountains. Conclusions:From 2014 to 2018, the compliance rates of fluoride in rural centralized water supply increase year by year, and some achievements have been made in fluorine reduction and water improvement project in Inner Mongolia. However, there are still some rural areas with low level of water fluoride compliance rates which mainly distribute in the central region of Inner Mongolia in the north of Yin Mountains and west of Greater Higgnan Mountains. The current focus of prevention and control should be shifted from "general control" to "precise fluorine control". In the future, it is necessary to implement treatment projects in key areas of fluorine pollution from the aspects of policy implementation and technological innovation to ensure the drinking water safety of local rural residents.

Simultaneous determination of verapamil and its metabolite norverapamil in human plasma by liquid chromatography-tandem mass spectrometry / 药学学报

A sensitive and simple high-performance liquid chromatographic tandem mass spectrometric (LC-MS/MS) method for the determination of verapamil and norverapamil in human plasma was established and utilized in a pharmacokinetic study in healthy patients. Protein was precipitated by methanol in plasma samples, and the analytes and internal standard were separated on an Agilent Zorbax Eclipse C18 column (50 mm×4.6 mm, 5 μm) with a gradient procedure using methanol-acetonitrile (50∶50) as the organic phase and 0.1% formic acid - 5% acetonitrile - 10 mmol·L-1 ammonium formate solution as the mobile phase at flow rate of 0.5 mL·min-1. Electrospray ionization (ESI) and multiple reaction monitoring (MRM) detection modes were used for quantitative detection of verapamil, norverapamil and verapamil-d6 (IS). In the mode of multiple reaction monitoring of positive-ions, the monitoring ion pairs of verapamil, norverapamil and the verapamil-d6 were m/z 445.0→165.2, m/z 441.0→165.2 and m/z 461.1→165.2, respectively. The quantitative lower limit (LLOQ) for the determination of verapamil and norverapamil concentrations in human plasma can reach 0.1 ng·mL-1 in this assay. The calibration curve concentration ranged from 0.1 to 50 ng·mL-1 with high linearity (r2 > 0.997). The matrix effect of verapamil and norverapamil was 99.2%-100% and 101%-102%, respectively. The recovery of verapamil and norverapamil was 86.8%-95.9% and 87.4%-94.8%, respectively. This method has good specificity and high sensitivity. The determination of the verapamil and norverapamil was not subject to the matrix effect and stable extraction recovery was achieved in this assay. This method could be used to determine the concentration of verapamil and norverapamil in human plasma and suitable for human pharmacokinetic studies after approved by ethics committee.

Effect of Sanggenone C on MC3T3-E1 Based on Molecular Docking Technology / 中国实验方剂学杂志

Objective::To observe the effect of sanggenone C (SanC) on the proliferation and differentiation of mouse MC3T3-E1 osteoblasts induced by dexamethasone (DEX), and to explore its mechanism. Method::Molecular docking was conducted between SanC and Runt-associated transcription factor 2(Runx2) protein structure obtained by homologous modeling. MC3T3-E1 cells were jointly treated by different concentrations of SanC (8, 16, and 32 μmol·L-1) and 1 μmol·L-1 DEX, and then cell counting kit-8(CCK-8) method was used to detect the effect of SanC on the proliferation of MC3T3-E1 osteoblasts. The alkaline phosphatase (ALP) activity of MC3T3-E1 osteoblasts was determined by reagent kit and the formation of mineralized bone nodules were detected by alizarin red staining. Real-time fluorescent quantitative polymerase chain reaction (Real-time PCR) was used to detect the mRNA expression of Runx2, ALP and Osterix. The protein expression of Runx2 was detected by Western blot. Result::The docking score of SanC and Runx2 was -9.78.As compared with the normal group, DEX group significantly reduced the cell survival rate (P<0.01), and the greatest difference occurred on the seventh day. As compared with DEX group, SanC could significantly promote the cell proliferation of MC3T3-E1 (P<0.01), in which 32 μmol·L-1 SanC had the largest difference in proliferation rate on seventh day. As compared with the normal group, the expression of Runx2, ALP and Osterix mRNA increased to a certain extent in DEX group(P<0.01). As compared with DEX group, the expression levels of Runx2, ALP and Osterix mRNA were up-regulated in different concentration groups of SanC in a dose-dependent manner (P<0.01). As compared with the normal group, the expression of Runx2 protein in DEX group decreased significantly (P<0.05), and as compared with DEX group, the expression of Runx2 protein in cells under the intervention of SanC increased significantly (P<0.01). Conclusion::SanC can promote the proliferation, differentiation and mineralization of MC3T3-E1 osteoblasts, and the mechanism may be related to the up-regulation of Runx2 expression.

Antibacterial activity and mechanism of moxifloxacin nanoparticles against drug-resistant Pseudomonas aeruginosa / 药学学报

We prepared moxifloxacin (MXF) loaded nanoparticles by nano-precipitation/self-assembly method, then compared the antibacterial activity of MXF and MXF loaded nanoparticles, and investigated the antibacterial mechanism of MXF loaded nanoparticles against Pseudomonas aeruginosa in vitro. The physicochemical properties such as particle size and zeta potential were investigated by laser particle size analyzer. The in vitro release characteristics were investigated by high performance liquid chromatography (HPLC). The effect of nanoparticles on HBE cells viability was investigated by CCK-8 assay. In addition, the in vitro antibacterial activity was investigated by minimum inhibitory concentration (MIC) assay, biofilm formation assays and transmission electron microscope (TEM) observation, then the antibacterial mechanism was initially explored. The particle size measurement showed that the nanoparticles had a size of 332.5 ± 2.7 nm, a polymer dispersion index (PDI) of 0.125 ± 0.053, a zeta potential of -24.3 ± 1.7 mV, and a uniform particle size distribution, drug loading content was (6.02 ± 1.27) %, encapsulation efficiency was (16.69 ± 1.17) %. The TEM results show that the nanoparticles have a spheroidal structure, and the particle size and distribution are consistent with the particle size measurement results. The nanoparticles can be effectively and rapidly released in phosphate buffer saline (PBS), releasing about 70% in 24 h, and releasing 87% in 72 h, and almost completely releasing the MXF at 120 h. At the same time, compared with moxifloxacin free drug, its MIC value is 8 μg·mL-1, which is 1/2 of MXF solution, and can significantly inhibit the formation of bacterial biofilms. It has well antibacterial activity in vitro and can be targeted to the surface of bacteria to exert its efficacy and improve the antibacterial effect. The moxifloxacin nanoparticles prepared in this study has a uniform particle size distribution, well drug release performance and antibacterial effect, and provides new ideas and strategies for the treatment of bacterial lung infection and the development of new antibacterial nanoformulations.

Effects of dezocine-remifentanil intravenous anaesthesia on perioperative signs, serum TNF-alpha and IL-6 in liver cancer patients undergoing radiofrequency ablation

Objective: To analyse the impact of dezocine-remifentanil intravenous anaesthesia on perioperative signs, serum tumour necrosis factor-alpha [TNF-alpha], and interleukin-6 [IL-6] in liver cancer patients undergoing radiofrequency ablation [RFA]

Study Design: An experimental study

Place And Duration Of Study: Renmin Hospital of Wuhan University, Wuhan, China, from January 2017 to February 2018

Methodology: Eighty patients with small hepatocellular carcinoma [SHCC] were selected as the research object. They were divided into Group A and Group B with the random number table method, with 40 cases in each group. Group A were given dezocine-remifentanil intravenous anaesthesia and Group B were given midazolam-remifentanil intravenous anaesthesia. Patients' situations in the surgery were compared between the two groups. Changes in heart rate [HR], mean arterial pressure [MAP] and blood oxygen saturation [SpO₂] were recorded before the surgery [T₀], at 5 minutes after the RFA [T₁] and at the end of the RFA [T₂]. Levels of tumour necrosis factor-alpha; [TNF-alpha;] and interleukin-6 [IL-6] on the 1² day after the RFA were compared between the two groups

Results: The wake-up time in Group A was shorter than Group B [p<0.001], and the VAS pain score in Group A was lower than Group B [p<0.001]. At T₁, the MAP in Group A was higher than Group B [p<0.001]. There was no significant difference in MAP between the two groups at T₀ and T₂ [p=0.881, 0.696, respectively]. At T₁ and T₂, the HR in Group A was lower than Group B [all p<0.001]. There was no significant difference in HR between the two groups at T₀ [p=0.684]. There was no significant difference in SpO₂ between the two groups at T₀, T₁ and T₂ [p=0.654, 0.884 and 0.798, respectively]. On the 1st day after the RFA, the level of TNF-alpha;, IL-6 in Group A were lower than those of Group B [all p<0.001]. There was no significant difference in the incidence of intraoperative complications between the two groups [p=0.644]

Conclusion: Compared with midazolam-remifentanil intravenous anaesthesia, the dezocine-remifentanil method has a better analgesic effect, shorter wake-up time, and can effectively regulate the expression of inflammatory cytokines TNF-alpha; and IL-6. However, the effect of remifentanil on the respiratory function is dose-dependent. Therefore, respiratory cycle monitoring and management should be strengthened during the surgery

Protective effect of Mori Cortex on kidney in rats with hyperlipidemia and hyperuricemia based on molecular docking technique / 中草药

Objective To investigate the mechanism of total flavonoids of Mori Cortex (TFMC) in improving hyperlipidemia and hyperuricemia related nephropathy. Methods The molecular structure of URAT1 protein and structure of Mori Cortex total flavonoids extract were docked by selecting the effective components of total flavonoids extract of Mori Cortex and related genes of uric acid. Forty SD rats were randomly divided into four groups: normal group, model group, TFMC group (1.0 g/kg) and benzbromarone group (6.25 mg/kg). The hyperlipidemia and hyperuricemia model was established by feeding with high fat diet plus adenine and ethylamine butanol. After 16 weeks, the levels of blood glucose and blood lipids in serum, uric acid (UA), creatinine (CRE), and urea nitrogen (BUN) of rats in each group were compared. The renal pathological changes were observed by hematoxylin eosin staining. The expressions of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), uric acid anion exchange transporter 1 (URAT1) and organic anion transporter 1 (OAT1) were detected by qRT-PCR. Results The main component of moracin of total flavonoids of Mori Cortex was the high target of the genes related to uric acid, suggesting that the moracin might be the main active component in the improvement of hyperlipidemia and hyperuricemia related nephropathy. After 16 weeks of drug intervention, the serum levels of Glu, TC, TG, LDL-C, UA, CRE and BUN in the model group were significantly higher than those in the normal group, and the level of HDL-C in the model group was significantly higher than that in the normal group (P < 0.05, 0.01). Compared with the model group, the above biochemical indexes in the TFMC group and the benzbromarone group were significantly decreased and HDL-C was significantly increased (P < 0.05, 0.01). The results of HE staining showed that the epithelial cells of renal tubules in the model group were swollen and necrotized, and the protein tubules could be seen in the renal tubules. In the TFMC group, some renal tubular epithelial cells were slightly swollen, and no inflammatory cells infiltrated around them. The structures of renal cortex and medullary were clear, and no hyperplasia or atrophy in glomerular were found, no tubular type and inflammatory cell infiltration in renal interstitial tissue of rats in benzbrommarone group were observed. The results of qRT-PCR showed that, compared with the normal group, the content of IL-6, TNF-α, and URAT1 mRNA was significantly increased, the content of OAT1 mRNA was significantly decreased in the model group; The content of above indicators was decreased and OAT1 was increased after drug intervention, (P < 0.05, 0.01). Conclusion The improvement of hyperlipidemia and hyperuricemia associated nephropathy may be related to the regulation of IL-6, TNF-α, URAT1, and OAT1 mRNA. Mori Cortex has obvious influence on the key factor of hyperlipidemia and hyperuricemia with URAT1 as its potential target, and the results of molecular virtual docking and animal experiments are similar. It provides a theoretical basis for further study on the improvement of hyperlipidemia and hyperuricemia related nephropathy and provides a reference for the further molecular mechanism.

Quantification of norfloxacin in human plasma by LC-MS/MS and validation of the analytical method / 药学学报

A LC-MS/MS method for quantification of norfloxacin in human plasma had been developed. This method was applied to the pharmacokinetics study of norfloxacin in the human. The plasma sample was precipitated by methanol and ciprofloxacin was used as the internal standard (IS). Chromatographic separation was performed on a Symmetry® C18 column(100 mm×4.6 mm, 5 μm). Mobile phase contains 0.3% formic acid and 5% methanol in deionized water at a flow rate of 0.45 mL·min-1. Norfloxacin and ciprofloxacin (IS) were ionized with an ESI source and operated in positive ion mode. The detected ions were m/z 320.3→302.1 (norfloxacin), m/z 332.3→314.1 (ciprofloxacin). This LC-MS/MS method yielded a linearity over the range of 10-1 000 ng·mL-1 with the lower limit of quantitation (LLOQ) of 10 ng·mL-1. The intra and inter-assay precisions (RSD%) were within the range of 2.64%-7.23% and the accuracy (RE%) was less than ±5.00%. The pharmacokinetic parameters tmax, Cmax, AUC0-t, and t1/2 were 1.28±0.364 h, 627±171 ng·mL-1, 2 938±850 h·ng·mL-1, and 6.01±1.36 h, respectively. This LC-MS /MS method was proven simple, sensitive, rapid and suitable for pharmacokinetics study of norfloxacin in the human and Approved by the Medical Ethics Committee of Liuzhou Workers' Hospital.

Endothelial glycocalyx as a potential theriapeutic target in organ injuries / 中华医学杂志(英文版)

OBJECTIVE@#The endothelial glycocalyx (eGC) is a dynamic and multicomponent layer of macromolecules found at the surface of vascular endothelium, which is largely underappreciated. It has recently been recognized that eGC is a major regulator of endothelial function and may have therapeutic value in organ injuries. This study aimed to explore the role of the eGC in various pathologic and physiologic conditions, by reviewing the basic research findings pertaining to the detection of the eGC and its clinical significance. We also explored different pharmacologic agents used to protect and rebuild the eGC.@*DATA SOURCES@#An in-depth search was performed in the PubMed database, focusing on research published after 2003 with keywords including eGC, permeability, glycocalyx and injuries, and glycocalyx protection.@*STUDY SELECTION@#Several authoritative reviews and original studies were identified and reviewed to summarize the characteristics of the eGC under physiologic and pathologic conditions as well as the detection and protection of the eGC.@*RESULTS@#The eGC degradation is closely associated with pathophysiologic changes such as vascular permeability, edema formation, mechanotransduction, and clotting cascade, together with neutrophil and platelet adhesion in diverse injury and disease states including inflammation (sepsis and trauma), ischemia-reperfusion injury, shock, hypervolemia, hypertension, hyperglycemia, and high Na as well as diabetes and atherosclerosis. Therapeutic strategies for protecting and rebuilding the eGC should be explored through experimental test and clinical verifications.@*CONCLUSIONS@#Disturbance of the eGC usually occurs at early stages of various clinical pathophysiologies which can be partly prevented and reversed by protecting and restoring the eGC. The eGC seems to be a promising diagnostic biomarker and therapeutic target in clinical settings.

Metabolic Characteristics of WBC-deplted RBC suspension during Different Storage Stage in MAP Based on UPLC-MS/MS / 中国实验血液学杂志

OBJECTIVE@#To explore the metabolic characteristics and metabolic markers of WBC-depleted RBCs in MAP preservation solution and to analyzed the metabolic profile of RBC in MAP preservation solution by using metabolomics.@*METHODS@#The changes of metabolitcs in 10 U WBC-depleted RBC suspension at 3-different storage period (D 0, D 14 and D 35) were detected by using the UPLC-MS/MS, the charaeteristic ions and metabolic markers of RBC stored in preservation sblution for 0 d, 14 d and 35 days were analyzed by using the principal component analysis(PCA).@*RESULTS@#The number of characteristic ions in RBC and supernatant extracts detected during the initial, middle and final storage could be clearly distinguiseed. The 5 metabolism-related substamces such as lact-c acid, nicotinamide, glucose, 5-htdroxyproline and malic acid showed statistically significant difference in 3 storage period.@*CONCLUSION@#The UPLC-MS/MS method combined with statistical analysis of multivariate data can be used to study the metabolic characteristics of RBC, the different metabolites of RBC in different storage stages can be used as the potential markers for evaluation of guality of RBC in storag period. The results of this study provide a basis for studing the RBC guality changes in storage period.

Comparison study between minimally invasive transforaminal lumbar interbody fusion via Quadrant minimally invasive system and open transforaminal lumbar interboday fusion in treating lumbar degeneration disease / 新乡医学院学报

Objective To compare the clinical effect between minimally invasive transforaminal lumbar interbody fusion (MIS-TLIF) via Quadrant minimally invasive system and open transforaminal lumbar interboday fusion(TLIF) in treating lumbar degeneration disease.Methods Clinical data of 46 patients in the First Affiliated Hospital of Xinxiang Medical University from April 2016 to April 2017 with lumbar degenerative diseases who were given operation treatment were analyzed retrospectively.Twenty patients underwent MIS-TLIF via Quadrant minimally invasive system(MIS-TLIF group) and twenty-six patients underwent open-TLIF(open TLIF group).The blood loss,operative time,length of incision,volume of drainage after operation,time of ambulation,hospitalization time,hospitalization cost,postoperative complications,visual pain simulation (VAS) score and Oswestry disability index(ODI) at 1,3,6 and 12 months after operation were compared between the two groups.Results The blood loss,length of incision,volume of drainage after operation,time of ambulation,hospitalization time,hospitalization cost in the MIS-TLIF group were less than those in the open TLIF group (P ＜0.05);there was no significant difference in operative time between the two groups(P ＜0.05).The VAS score and ODI of the MIS-TLIF group were lower than those of the open TLIF group at 1 and 3 months after surgery (P ＜ 0.05);there was no significant difference in the VAS score and ODI between the two groups at 6 and 12 months after surgery (P ＜ 0.05).There was no complication such as dural and nerve injury and internal fixation rupture in the two groups.In the open TLIF group,postoperative wound healing was delayed in two cases,postoperative incision infection occurred in one cases,the incidence of postoperative complication was 11.5％ (3/26).In the MIS-TLIF group no postoperative complication occurred,the incidence of postoperative complication was 0.0％ (0/20);the incidence of postoperative complications was not statistically significant between the two groups (x2 =2.471,P ＜ 0.05).Conclusion MIS-TLIF via Quadrant minimally invasive system has the advantages of less blood loss,incision and volume of drainage after operation in treating lumbar degenerative disease,and it can be used as the first choice for the treatment of lumbar degenerative disease.