Cloning and Expression Analysis of Squalene Epoxidase Gene in Poria cocos / 中国实验方剂学杂志

ObjectiveTo clone squalene epoxidase （SE）， a potential key rate-limiting enzyme involved in the synthesis pathway of Poria cocos triterpenes， from P. cocos and analyze for bioinformatics and expression. MethodThe total RNA was extracted by the kit and reverse-transcribed to cDNA. Specific primers were designed， and the cDNA was used as a template for cloning the SE gene， which was analyzed for bioinformatics. The expression of P. cocos qualene epoxidase（PcSE） was examined by Real-time polymerase chain reaction（Real-time PCR） in P. coco Shenzhou No. 10， Xiangjing 28， and 5.78 strains. ResultThe full length of PcSE is 1 571 bp， containing four exons and three introns. The obtained CDS sequence is 1 413 bp， encoding 470 amino acids. This protein is a hydrophobic protein with no signal peptide structure and has two transmembrane structural domains with a FAD/NAD （P） binding domain and SE structural domain localized to the mitochondrial membrane and the plasma membrane. The homologous sequence alignment with fungi of the Poriferae family is 80.92%， and the phylogenetic tree shows that PcSE protein is most closely related to P. cocos from the US. The results of Real-time PCR showed that the PcSE was expressed in all three strains， with the highest expression in 5.78 strain， and there was no significant difference in PcSE expression among the three strains. ConclusionFor the first time， the PcSE gene was cloned and analyzed from P. cocos， providing a basis for further research on the function of PcSE and the analysis of P. cocos triterpene biosynthesis pathway.

Syndrome Differentiation and Treatment of Obesity from "Orchids to Eliminate Stagnant Qi" / 中国实验方剂学杂志

Obesity is a chronic， recurrent， and progressive metabolic disorder characterized by the abnormal or excessive accumulation of body fat caused by multiple factors such as genetics， dietary structure， lifestyle and behavior， psychology， environment， and society， leading to an energy surplus. Obesity is a major risk factor that increases the risk of developing various chronic diseases， including type 2 diabetes， hypertension， cardiovascular diseases， stroke， and certain malignancies. The global incidence of obesity is increasing year by year. With the continuous improvement of people's living standards， more than half of adults in China are now overweight or obese， posing a serious threat to people's health and increasing the social and economic burden. It has become a pressing major public health issue that needs to be addressed urgently. The concept of obesity can be traced back to the Huangdi’s Internal Classic （Huang Di Nei Jing）， which describes it as "the problems in fat and affluent people are caused by excessive taking of rich food"， and suggests that ''frequent intake of rich and greasy foods can produce interior heat. Sweet flavor causes chest fullness. That is why its spleen-Qi flows upwards and changes into consumption-thirst disease. It can be treated by Eupatorii Herba which is used to remove stagnant Qi''. The stagnant qi is caused by the transformation failure of rich and greasy food and wine， so obesity is the disease of stagnant qi. Obesity is caused by indulging in rich and greasy food， wine， spicy and flavorful foods， raw and cold foods， and sweet and greasy foods， or overeating and leading a sedentary lifestyle， staying up late， or experiencing emotional imbalances such as excessive joy， anger， worry， pensiveness， and fear. It can also be caused by congenital abnormalities， leading to improper functioning of the spleen and stomach， dysregulation of the absorption and secretion of the small intestine， and the accumulation of stagnant Qi in the organs and muscles， resulting in a plump physique. The intake of food and drink depends on the functions of the stomach in receiving and decomposing， the small intestine in absorbing and secreting， and the spleen in transforming and transporting. The affected organs in obesity are the spleen， stomach， and small intestine. Orchids， specifically Eupatorii Herba and Lycopi Herba， are aromatic herbs that can regulate the smooth flow of Qi， eliminate stagnation， and cleanse impurities. In a broader sense， any aromatic and pungent substance that can invigorate the spleen， promote clarity， harmonize the stomach， reduce turbidity， and assist in the normal secretion and absorption functions of the small intestine， thereby eliminating excess， is referred to as orchid. Therefore， the treatment principle for obesity is to use ''orchids to eliminate stagnant Qi''， aiming to regulate the functions of the spleen， stomach， and small intestine using aromatic and pungent substances， gradually eliminating excessive dampness， phlegm， turbidity， and heat， and restore the balance of the middle energizer. This way， individuals who are obese can achieve a non-obese state.

Development and construction of comprehensive examination database for periodontology based on curriculum integration / 中华医学教育探索杂志

Periodontology is one of the important disciplines in oral clinical medicine, which covers a wide range of subjects and intersects with many basic disciplines. Under the environment of the implementation of modular teaching in Shanghai Jiao Tong University School of Medicine, the assessment method with separate propositions for the teaching and research section is still adopted. There is a mismatch between the assessment mode and the curriculum setting; the basic subject propositions are difficult to be combined with clinical cases; the knowledge point assessment is single, and the students' ability to integrate the knowledge points cannot be assessed. The development and construction of the comprehensive examination database for periodontology was based on curriculum integration, gathering the teaching backbones of various disciplines, focusing on periodontology, radiating all related disciplines, unifying the proposition outline, proposition type, proposition principle, combining with relevant knowledge points of various disciplines based on clinical cases, and tried to apply to clinical students majoring in stomatology. The use of the examination database promotes students' ability to flexibly apply theoretical knowledge to clinical case analysis, further promotes the reform of modular teaching, lays a solid foundation for future clinical work, and meanwhile provides an important basis for directions of the teaching and research section.

Value of neuron specific enolase and S-100β protein in cerebrospinal fluid in the diagnosis and treatment of children with viral encephalitis and meningitis / 中国基层医药

Objective:To investigate the changes and clinical significance of neuron specific enolase (NSE) and S-100β protein levels in cerebrospinal fluid of children with viral encephalitis and meningitis.Methods:Sixty children with viral encephalitis and meningitis admitted to The Second Hospital of Jiaxing from February 2018 to December 2020 were included in the observation group. An additional 30 children without central nervous system diseases who concurrently received treatment in the same hospital were included in the control group. The value of NSE and S-100β protein levels in the diagnosis and treatment of viral encephalitis and meningitis in chiblren were analyzed.Results:NSE and S-100β protein levels in the observation group were (17.683 ± 1.321) μg/L and (1.755 ± 0.129) μg/L, respectively, which were significantly higher than (5.267 ± 0.907) μg/L and (0.827 ± 0.172) μg/L in the control group ( t = 46.25, 28.65, both P < 0.001). NSE and S-100β protein levels in children with mild viral encephalitis and meningitis were (15.219 ± 0.870) μg/L and (1.456 ± 0.113) μg/L, respectively, which were significantly lower than (19.893 ± 1.066) μg/L and (2.014 ± 0.085) μg/L in children with severe viral encephalitis and meningitis ( t = -18.69, -21.32, both P < 0.001). In children with viral encephalitis and meningitis, NSE and S-100β protein levels during the acute phase were (17.250 ± 1.188) μg/L and (1.683 ± 0.096) μg/L, respectively, which were significantly higher than (11.150 ± 0.971) μg/L and (1.147 ± 0.098) μg/L during the convalescence phase ( t = 30.79, 30.27, both P < 0.001). Conclusion:NSE and S-100β protein levels in cerebrospinal fluid of children with viral encephalitis and meningitis can help evaluate the severity of viral encephalitis and meningitis in children, providing important clinical application value for judging the development and prognosis of viral encephalitis and meningitis.

Evaluation on Surgical Robotic Pose Repeatability Based on Laser Tracking Measurement / 中国医疗器械杂志

To provide accurate information for registration and safety evaluation of surgical robot, the pose repeatability measurement method was proposed. According to the terminal instrument of the master-slave surgical robot (such as high-frequency electric knife, ultrasonic knife), a suitable target ball fixture was designed. The node data at 10%, 50% and 100% rated speed were measured respectively. Through data analysis, the pose repeatability property of the tested samples at different speeds was obtained. It has high applicability and repeatability, and can meet the requirements of data traceability and registration testing.

Construction and practice of CRC management system based on the whole process of clinical trial projects / 中华医学科研管理杂志

Objective:To introduce the construction and practice of the Clinical Research Coordinator(CRC) management system based on the whole process of the clinical trial project in Ninth People′s Hospital of Shanghai Jiao Tong University School of Medicine.Methods:Constructed CRC management system with hospital features, including the management of admission, examination, training, emergency, and evaluation.Results:CRC management system was put into practice, and investigators, sponsors, and drug clinical trial institutions were highly satisfied with clinical trials using this system.Conclusions:With the gradual formation of CRC industry norms and consensus, the standardized management of CRC can promote the development of China′s pharmaceutical industry.

Establishment of random forest model and its predictive value for pathologic complete response in breast cancer patients undergoing neoadjuvant chemotherapy / 肿瘤研究与临床

Objective:To investigate the predictive value of established random forest model for pathologic complete response (pCR) in breast cancer patients undergoing neoadjuvant chemotherapy.Methods:The clinicopathologic data of 142 primary breast cancer patients undergoing breast-conserving surgery or modified radical mastectomy after neoadjuvant chemotherapy from Cangzhou Central Hospital between January 2010 and October 2021 were retrospectively analyzed. Histologically, breast and axillary lymph node without residual infiltrated tumors was treated as pCR. The patients were divided into pCR group (23 cases) and non-pCR group (119 cases) according to whether patients achieved pCR or not, and the differences of clinicopathologic data between the two groups were compared. The risk factors affecting pCR were identified by using logistic regression analysis, random forest model was established by using random forest function of R statistical software, and Gini index of random forest algorithmic was used to order the importance of variables. The receiver operating characteristic (ROC) curve was used to assess the value of random forest model in predicting the efficacy of neoadjuvant chemotherapy.Results:The overall pCR ratio after neoadjuvant chemotherapy was 16.20% (23/142). The proportion of tumor diameter ≤5 cm, negative axillary lymph node, negative human epidermal growth factor receptor 2 (HER2), Ki-67 positive index >20%, histological grade 2, and neoadjuvant chemotherapy regimens including targeted therapy in pCR group was higher than that in non-pCR group, and the difference was statistically significant (all P < 0.05). Univariate logistic regression analysis showed that tumor diameter, axillary lymph node, HER2, Ki-67, histological grade, and neoadjuvant chemotherapy regimens were related with pCR (all P < 0.05). Multivariate logistic regression analysis showed that tumor diameter >5 cm ( OR = 5.85, 95% CI 1.28-26.67, P = 0.022), positive axillary lymph node ( OR = 11.22, 95% CI 1.84-68.42, P = 0.009), positive HER2 ( OR = 7.35, 95% CI 1.45-37.26, P = 0.016), Ki-67 positive index ≤20% ( OR = 1.03, 95% CI 1.01-1.06, P = 0.017), histological grade 3 ( OR = 7.37, 95% CI 1.24-43.86, P = 0.028), and non-targeted therapy ( OR = 0.02, 95% CI 0.00-0.25, P = 0.003) were independent risk factors of pCR. Random forest algorithm showed that the importance order of risk factors of pCR was successively Ki-67 low expression, positive axillary lymph node, tumor diameter >5 cm, positive HER2, non-targeted therapy and histological grade 3. The area under the ROC curve of random forest model for predicting pCR was 0.84 (95% CI 0.74-0.93); the sensitivity was 87.0% and specificity was 72.3% when the optimal cut-off value was 0.88. Conclusions:Low expression of Ki-67, positive axillary lymph node, tumor diameter >5cm, positive HER2, non-targeted therapy and histological grade 3 are risk factors of pCR in breast cancer patients after neoadjuvant chemotheapy. Random forest model helps to predict pCR in breast cancer patients after neoadjuvant chemotheapy.

Predictive value of systemic inflammation response index before treatment for pathological complete response in patients with breast cancer undergoing neoadjuvant chemotherapy / 国际肿瘤学杂志

Objective:To investigate the predictive value of systemic inflammation response index (SIRI) before treatment for pathological complete response (pCR) in patients with breast cancer undergoing neoadjuvant chemotherapy.Methods:The clinicopathological data of 119 patients with primary breast cancer undergoing neoadjuvant chemotherapy and subsequent breast-conserving or modified radical surgery from Cangzhou Central Hospital of Hebei Province between January 2010 to March 2020 were retrospectively analyzed, and patients were divided into pCR group ( n=19) and non-pCR group ( n=100) based on postoperative pathology. The SIRI before treatment between the two groups was compared. The patients were divided into SIRI≤0.25 ( n=10) , 0.26-0.50 ( n=42) , 0.51-0.75 ( n=29) , 0.76-1.00 ( n=19) , and >1.00 ( n=19) groups according the SIRI before treatment, and the pCR ratios of the five groups were compared. Spearman correlation analysis was applied to evaluate the relationship between SIRI before treatment and pCR, logistic regression analysis was used to identify the influencing factors of pCR for neoadjuvant chemotherapy in breast cancer patients, and receiver operating characteristic (ROC) curve was used to evaluate the predictive value of SIRI before treatment for pCR of neoadjuvant chemotherapy in breast cancer patients. Results:Tumor size ( Z=2.26, P=0.024) , axillary lymph node metastasis ( χ2=5.73, P=0.017) , human epidermal growth factor receptor-2 (HER-2) ( χ2=8.77, P=0.003) , Ki-67 ( Z=2.68, P=0.007) , cytological nuclear grade ( χ2=5.08, P=0.024) , neutrophil count before treatment ( Z=2.44, P=0.015) , monocyte/lymphocyte ratio before treatment ( Z=3.04, P=0.002) , and SIRI before treatment ( Z=3.29, P=0.001) had statistical differences between the pCR and non-pCR groups. The pCR ratios were 50% (5/10) in the SIRI ≤0.25 group, 21% (9/42) in the 0.26-0.50 group, 10% (3/29) in the 0.51-0.75 group, 11% (2/19) in the 0.76-1.00 group, and 0 (0/19) in the >1.00 group, with a statistic difference ( χ2=14.28, P=0.006) . SIRI before treatment was negatively related with pCR ( r=-0.30, P=0.001) . Univariate logistic regression analysis showed that tumor size ( OR=0.50, 95% CI: 0.28-0.89, P=0.019) , axillary lymph node metastasis ( OR=5.43, 95% CI: 1.19-24.83, P=0.029) , HER-2 ( OR=7.54, 95% CI: 1.65-34.36, P=0.009) , Ki-67 ( OR=1.03, 95% CI: 1.01-1.05, P=0.008) , cytological nuclear grade ( OR=0.20, 95% CI: 0.04-0.92, P=0.038) , neutrophil count before treatment ( OR=0.54, 95% CI: 0.32-0.92, P=0.023) , monocyte/lymphocyte ratio before treatment ( OR=0.00, 95% CI: 0.00-0.01, P=0.007) , and SIRI before treatment ( OR=0.03, 95% CI: 0.00-0.37, P=0.007) were influencing factors for pCR of neoadjuvant chemotherapy in breast cancer patients. Multivariate logistic regression analysis confirmed that tumor size ( OR=0.31, 95% CI: 0.14-0.72, P=0.007) , axillary lymph node metastasis ( OR=10.97, 95% CI: 1.35-89.61, P=0.025) , HER-2 ( OR=6.47, 95% CI: 1.18-35.65, P=0.032) , Ki-67 ( OR=1.04, 95% CI: 1.00-1.07, P=0.029) , cytological nuclear grade ( OR=7.87, 95% CI: 1.01-61.35, P=0.049) , and SIRI before treatment ( OR=0.03, 95% CI: 0.00-0.58, P=0.020) were independent influencing factors for pCR of neoadjuvant chemotherapy in breast cancer patients. The ROC curve showed that the area under the curve of SIRI before treatment for predicting pCR was 0.74 (95% CI: 0.65-0.82) , sensitivity was 68.0%, and specificity was 75.3%. The area under the curve of monocyte/lymphocyte ratio before treatment for predicting pCR was 0.72 (95% CI: 0.63-0.80) , sensitivity was 48.0%, and specificity was 84.2%. The area under the curve of neutrophil count before treatment for predicting pCR was 0.68 (95% CI: 0.59-0.76) , sensitivity was 61.0%, and specificity was 83.7%. Conclusion:SIRI before treatment may serve as a marker for predicting pCR in patients with breast cancer undergoing neoadjuvant chemotherapy, patients with low SIRI are more likely to obtain pCR.

Site-specific integration and stable expression of exogenous protein at a novel site on CHO cell chromosome / 中国药科大学学报

@#Finding stable expression sites on the chromosomes of Chinese hamster ovary (CHO) cells is an effective method to solve the problem of unstable expression of CHO cells in long-term culture. Our group used lentiviral transfection to integrate the tracer gene (Zsgreen1) into the chromosome of CHO cells and found multiple potential stable expression sites. This study verified the ability of one of the sites located in the 148052-148157 bp region on chromosome NW_003614241.1 to stably express exogenous proteins.The expression of Zsgreen1 gene was first observed, and CRISPR/Cas9 technology was then used to integrate the enhanced green fluorescent protein (EGFP) gene into this site. Three strains of EGFP gene integrated cells were obtained. After 60 generations of suspension culture, the fluorescence intensity of the cells had no significant changes, which proved that this site can stably express the EGFP gene. The same method was used to construct recombinant CHO cell lines expressing the human serum albumin (HSA) gene, and was verified by Western blot that this site could express and secrete HSA. It shows that the above-mentioned sites can be integrated and can stably express exogenous proteins.

L-carnitine treatment attenuates renal tubulointerstitial fibrosis induced by unilateral ureteral obstruction

Background/Aims@#Accumulating evidence indicates that L-carnitine (LC) protects against multiorgan damage through its antioxidant properties and preservation of the mitochondria. Little information is available about the effects of LC on renal fibrosis. This study examined whether LC treatment would provide renoprotection in a rat model of unilateral ureteral obstruction (UUO) and in vitro. @*Methods@#Sprague-Dawley rats that underwent UUO were treated daily with LC for 7 or 14 days. The influence of LC on renal injury caused by UUO was evaluated by histopathology, and analysis of gene expression, oxidative stress, mitochondrial function, programmed cell death, and phosphatidylinositol 3-kinase (PI3K)/ AKT/forkhead box protein O 1a (FoxO1a) signaling. In addition, H2O2-exposed human kidney cells (HK-2) were treated with LC. @*Results@#LC treatment inhibited expression of proinflammatory and profibrotic cytokines, and was followed by a significant attenuation of tubulointerstitial inflammation and fibrosis. The increased oxidative stress caused by UUO was associated with mitochondrial dysfunction and excessive apoptosis and autophagy via PI3K/AKT/FoxO1a-dependent signaling, and this was abrogated by administration of LC. In H2O2-exposed HK-2 cells, LC decreased intracellular production of reactive oxygen species, and suppressed expression of profibrotic cytokines and reduced the number of apoptotic cells. @*Conclusions@#LC protects against the progression of tubulointerstitial fibrosis in an obstructed kidney.

Proteomics and metabolic phenotyping define principal roles for the aryl hydrocarbon receptor in mouse liver

Dioxin-like molecules have been associated with endocrine disruption and liver disease. To better understand aryl hydrocarbon receptor (AHR) biology, metabolic phenotyping and liver proteomics were performed in mice following ligand-activation or whole-body genetic ablation of this receptor. Male wild type (WT) and

Assessment of nephrotoxicity of herbal medicine containing aristolochic acid in mice

Background/Aims@#It is undetermined if herbal medicines (HM) containing aristolochic acid (AA)-containing have similar nephrotoxicity to AA itself. @*Methods@#We administered HM containing a high concentration of AA for 5 days (short-term study) or a low concentration of AA for 30 days (long-term study) to C57BL/6 mice; for comparison, same dose of AA compound was used as controls. @*Results@#The nephrotoxicity in the HM- and AA-treated mice was compared in terms of renal function, histopathology, oxidative stress, apoptotic cell death, and mitochondrial damage. Short-term HM treatment resulted in acute kidney injury (marked renal dysfunction, acute tubular necrosis, and neutrophil gelatinase-associated lipocalin [NGAL] expression) in which the severity of renal dysfunction and histopathology was comparable with that induced by the administration of AA alone. Long-term HM treatment resulted in features of chronic kidney disease (CKD, mild renal dysfunction and tubular atrophy and dilatation). No significant differences in these parameters were observed between the HM- and AA-treated mice. HM-induced oxidative stress (8-hydroxy-2’-deoxyguanosine and manganese- dependent superoxide dismutase expression) and apoptotic cell death (terminal deoxynucleotidyl transferase dUTP nick end labelling [TUNEL]-positive cells and active caspase-3 expression) were similar in HM- and AA-treated mice in the short-term and long-term studies. Mitochondrial injury, evaluated by electron microscopy, was also similar in HM- and AA-treated mice in the short-term and long-term studies. @*Conclusions@#The nephrotoxic potential of HM containing AA was similar to that of AA itself.

Determination of hydroxylsafflower yellow A in safflower W/O cream by RP-HPLC / 中国药科大学学报

@#A reversed phase HPLC method for determination of hydroxylsafflower yellow A in safflower W/O cream was established. The column was Zorbax Eclipse C18 column(4. 6 mm×250 mm, 5 μm), and the mobile phase was composed of methanol, acetonitrile and 0. 02% phosphoric acid solution(26 ∶2〓 ∶72). The flow rate of mobile phase was set at 1. 0 mL/min, and the column temperature was kept at 55 °C. The detection wavelength was 403 nm. Safflower W/O cream was successively demulsified with methanol at high temperature and followed by the addition of purified water for the extraction. The results showed that the excipients did not interfere with the chromatographic peak of hydroxylsafflower yellow A. Hydroxylsafflower yellow A presented a good linear relationship in the range of 1. 236- 12. 36 μg/mL(y=156. 17x+1. 198 3, r=0. 999 5), and the detection limit was 23. 6 ng/mL with the quantitative limit of 118 ng/mL. The percentage of extracting recovery was in the range of 99. 7% to 103. 3%. The precision RSD was 0. 12%(n=6), and the sample stability was acceptable when being stored at room temperature for 24 h. The developed method in this study was simple, rapid, accurate and reproducible, and can be used for the determination of hydroxylsafflower yellow A in safflower W/O cream.

Combined anatomic and physiologic scoring systems for predicting in-hospital mortality in ICU patients with severe trauma: A multicenter observational cohort study

Objective: To evaluate the ability of new injury severity score (NISS), acute physiology and chronic health evaluation II (APACHE II), Glasgow coma scale (GCS), a combination of NISS and GCS, a combination of APACHE II and GCS, a combination of NISS and APACHE II to predict all-cause mortality of patients with severe trauma in mainland China. Methods: This was a multicenter observational cohort study conducted in the ICU of the Chonggang General Hospital, Daping Hospital of the Army Medical University and Affiliated Hospital of Zunyi Medical College from January 2012 to August 2016. The score of NISS, APACHE II, GCS, a combination of NISS and GCS, a combination of APACHE II and GCS, a combination of NISS and APACHE II were calculated based on data from the first 24 hours of ICU admission. Data were processed with Student's t-test, chi-square test, and receiver operating characteristic (ROC) curve of six scoring systems. Calibration was assessed with the Hosmer-Lemeshow test. The primary endpoint was death from any cause during ICU stay. Results: A total of 852 and 238 patients with severe trauma were assigned to the derivation group and validation group, respectively. Area under the ROC curve (AUC) was 0.826 [95% confidence interval (CI)=0.794-0.855)] for NISS, 0.802 (95% CI=0.768-0.832) for APACHE II, 0.808 (95% CI=0.774-0.838) for NGCS, 0.859 (95% CI=0.829 -0.886) for NISS+NGCS, 0.864 (95% CI=0.835-0.890) for APACHE II +NGCS, 0.896 (95% CI=0.869-0.929) for NISS+APACHE II in the derivation cohort. Similarly, the score of NISS+APACHE II was also better than the other five scores in the validation cohort (AUC=0.782; 95% CI=0.725-0.833) and had a good calibration (P=0.41). Conclusions: Taking into account anatomical and physiological parameters completely, the combination of NISS and APACHE II performs better than NISS, APACHE II, NGCS, NISS+NGCS, APACHE II +NGCS for predicting mortality in ICU severe trauma patients. It is needful to develop models that contain various types of accessible predictors (demographic variables, injury cause/mechanism, physiological and anatomical variables, etc.) as comprehensive as possible.

Study on the bioactivity against hematologic malignancies and theoretical binding mechanism of a novel PI3K inhibitor JN-65 / 中国药科大学学报

@#The PI3K signaling pathway is frequently over-expressed in a variety of hematologic malignancies, so the development of PI3K inhibitors for the treatment of hematologic malignancies has broad application prospects. In this study, a novel PI3K inhibitor, JN-65, was identified through the investigation effects in the inhibition to hematologic malignancies. By MTT assays, JN-65 was found to effectively suppress the proliferation of hematologic malignancies, especially leukemia cell lines. The cell-free enzymatic studies demonstrated that JN-65 cloud inhibit PI3K and specifically inhibited PI3Kγ at low micromolar concentrations. Western blot confirmed that JN-65 could effectively inhibit the PI3K/Akt signaling pathway and the flow cytometry assays verified that JN-65 could induce the apoptosis of tumor cells through the suppression of PI3K signaling pathway. Finally, the molecular docking simulation method was used to explore the interaction between JN-65 and PI3K, and the inhibition mechanism of PI3K was revealed at the molecular level. In general, JN-65 would be a potential PI3K inhibitor for the treatment of hematologic malignancies.

Preparation and evaluation of an oral insulin enteric preparation based on polymer-lipid hybrid nanoparticles / 中国药科大学学报

@#To improve the oral bioavailability of insulin, an insulin-loaded enteric polymer-lipid hybrid nanoparticles(INS-NPs L100)was prepared using methoxy PEG-poly(D, L-lactide)(PEG-PLA), phospholipid s75 and Eudragit L100; in vitro and in vivo behaviors of INS-NPs L100 were evaluated. Insulin-loaded polymer-lipid hybrid nanoparticles(INS-NPs)were prepared by W/O/W double emulsion solvent evaporation method. INS-NPs formulation was optimized by single factor experiment using encapsulation efficiency, particle size, and in vitro release behavior of the corresponding INS-NPs L100 as evaluation indexes. The morphology, in vitro drug release profile and hypoglycemic effect of the INS-NPs L100 using the optimal INS-NPs and Eudragit® L100(used as enteric polymer)were assessed. The results showed that the encapsulation efficiency of the optimal INS-NPs was(62. 18±4. 51)%. The average particle size, PDI and Zeta potential was(225. 2±94. 3)nm, 0. 191±0. 068, and -(14. 84±1. 26)mV, respectively. The cumulative drug release from the INS-NPs L100 was only 8. 01% at 2 h in pH 1. 0 HCl solution, exhibiting a slow drug release behavior; while the drug release from INS-NPs L100 was 67. 31% at 6 h in phosphate buffer of pH 6. 8. Mereorer, after oral administration of INS-NPs L100 with a dose of 38 IU/kg, the blood glucose concentration of healthy rats was reduced to 76% of the initial values at 3. 5 h, exhibiting a sustained hypoglycemic effect. In summary, the INS-NPs L100 prepared in this study could effectively decrease the release rate of insulin in gastric juice, improve the stability of protein in the gastrointestinal tract, and provide a new approach for the oral administration of peptides and protein drugs.

Anti-liver fibrosis activities of the extracellular domain of transforming growth factor beta type II receptor fusion protein in vivo / 中国药科大学学报

@#This study is performed to analyze the anti-liver fibrosis effect of the fusion protein of human serum albumin and extracellular domain of transforming growth factor beta type II receptor(eTGFBR2)in vivo to looking for the more stable anti-liver fibrosis drug. The mice model of liver fibrosis was constructed by CCl4 induction and the following groups are included in the study: the control group, CCl4 model group, the positive control group, eTGFBR2 treatment group, HSA-eTGFBR2 treatment group, and HSA group. Hematoxylin eosin staining, serum liver function index detection, and western blot are used to identify the anti-liver fibrosis activities. The results showed that: (1)CCl4 caused liver structure disorder, hepatocellular necrosis, collagen fibers proliferation, and induced liver fibrosis at last; (2)HSA-eTGFBR2 and its monomer drug improved the symptoms of liver fibrosis significantly, as well as reduced the damage of liver cells and collagen deposition, and recovered the liver basic structure to normal. Both of HSA-eTGFBR2 and its monomer drug improved liver function and reduced the expression level of liver fibrosis marker α-SMA and COL I. Moreover, the anti-liver fibrosis effect of the fusion protein is comparable to the monomer drug. In contrast, the albumin had no effect on therapeutic effect; (3)Reducing the injection frequency of HSA-eTGFBR2 achieved the comparable effects to the monomer drug with the normal injection frequency. In summary, the fusion protein HSA-eTGFBR2 has good anti-liver fibrosis effect. In addition, reducing the injection frequency of the fusion protein could also achieve the comparable treatment with the monomer drug, indicating that the fusion protein is stable and has longer half-lives and then a relatively positive application prospect in future.

Recent Advances in the Development of Experimental Therapeutics for Levodopa-Induced Dyskinesia

No abstract available.

Risedronate inhibits rat bone marrow adipogenesis and reduces RANKL expression in adipocytes / 南方医科大学学报

OBJECTIVE@#To investigate the effects of risedronate on bone marrow adipogenesis and the expression of the receptor activator of nuclear factor κB ligand (RANKL) in adipocytes in the bone marrow micro-environment.@*METHODS@#Primary cultured rat mesenchymal stem cells (BMSCs) with or without adipogenic induction for 14 days were treated with 1, 5, 10, and 25 μmol/L risedronate. The droplets of the differentiated adipocytes were analyzed, and Western blotting was performed to detect the expression level of RANKL. Female SD rats (24-week-old) were randomly divided into sham-operated group and ovariectomy (OVX) group, and 12 weeks after the operation, the OVX rats were further divided into control group and risedronate group (2.4 μg/kg, injected subcutaneously for 3 times a week). Eight weeks later, the bone mineral density (BMD) of the rats and bone marrow histopathology of the femurs was examined to evaluate the effect of risedronate on the fat fraction in the bone marrow.@*RESULTS@#Risdronate significantly inhibited adipogenic differentiation of rat BMSCs and suppressed RANKL expression in the adipocytes derived from the BMSCs in a concentration-dependent manner. In OVX rats, risdronate treatment significantly increased the BMD and decreased the fat content in the bone marrow.@*CONCLUSIONS@#Risdronate can effectively inhibit the adipogenic differentiation of rat BMSCs, decrease fat content in the bone marrow, and suppress the generation and function of osteoclasts by down-regulating the expression of RANKL, which can be an important mechanism underlying the therapeutic effect of risedronate against osteoporosis.

Shen-Kang protects against tacrolimus-induced renal injury

BACKGROUND/AIMS@#Evidence suggests that Shen-Kang (SK), a traditional Chinese herbal medicine, protects against various types of renal injury. In this study, we evaluated whether SK treatment confers renoprotection in a rat model of chronic tacrolimus (TAC) nephropathy.@*METHODS@#Rats were treated daily with TAC (1.5mg/kg, subcutaneously) and SK (450 mg/kg, intravenously) for 4 weeks. The effects of SK on TAC-induced renal injury were assessed by measuring renal function, urine albumin excretion, histopathology, inflammatory cell infiltration, expression of profibrotic (transforming growth factor β1 [TGF-β1] and TGF-β inducible gene-h3 [βig-h3]) and proinflammatory cytokines, oxidative stress, and apoptotic cell death.@*RESULTS@#Administration of SK preserved glomerular integrity (fractional mesangial area and Wilms tumor 1-positive glomeruli), attenuated tubulointerstitial fibrosis, and reduced the number of ectodermal dysplasia 1-positive cells, and this was paralleled by improved urine albumin excretion and renal dysfunction. At the molecular level, SK treatment suppressed expression of TGF-β1/Smad2/3, βig-h3, and proinflammatory cytokines. Oxidative stress and apoptotic cell death were significantly decreased with SK treatment, and apoptosis-related genes were regulated toward cell survival (active caspase-3 and the B-cell lymphoma-2/Bcl2-associated X [Bcl-2/Bax] ratio).@*CONCLUSIONS@#SK protects against TAC-induced renal injury.