OBJECTIVE:To study the characteristics of patent behaviors of Chinese pharmaceutical demonstration enterprises for intellectual property,and to provide reference for the improvement of patent behaviors in Chinese pharmaceutial enterprises. METHODS:After retrieving the applicants(patentees)from China Intellectual Property Right Net on the basis of taking 7 pharma-ceutical demonstration enterprises chosen through appraisal by State Intellectual Property Office of the P.R.C in 2013 as the study samples,empirical study was conducted on their patent behaviors during 1985 and 2013,including the number and type of the pat-ents for which applications were made,international patent classification and legal status. RESULTS:The number of patent applica-tions of the 7 pharmaceutical demonstration enterprises demonstrated a growing trend(an annual increase up to 79.0%). The appli-cations were dominated by the inventive patent(84.9%). Many patents were in an authorized legal status(46.7%). All of these en-terprises paid more attention to the maintenance of the authorized patent. The technology patent applications were mainly about chemical compound,composition,and preparation methods,including the technical proposal containing the active ingredients and properties of the pharmaceutical preparation (A61K),particularly the application of the technical proposal related to the treatment with the preparation (A61P). CONCLUSIONS:All the 7 pharmaceutical demonstration enterprises were aware of patent competi-tion and protection and characterized by a higher proportion of the application for the inventive patent,perfect patent information management institution and high-quality patent applications. Chinese pharmaceutical enterprises can enhance their competitiveness and creativity through adjusting patent behaviors and increasing their awareness of patent competition.

Objective To assess the factors that influence the accuracy of real-time continuous glucose monitoring system (RT-CGM).Methods A total of 79 diabetic patients wore RT-CGM for three days continuously while calibrating by interphalangeal glucose values 4-8 times a day.We counted matching rate of interphalangeal glucose values and RT-CGM probe value,and analyzed correlation of the matching rate with MAGE,SDBG,MBG,AUC10,AUC3.9,and NGE by Pearson correlation analysis and multiple linear regression analysis.Then we grouped corresponding match rate by MAGE and MBG,compared the difference among groups.Furthermore,we analyzed correlation of RT-CGM probe values of all-day,fasting-postprandial,and nighttime and the corresponding interphalangeal glucose values by Pearson correlation analysis.Results Pearson correlation analysis showed that the matching rate was negatively correlated with the MAGE,SDBG,MBG,and AUC10.MAGE,MBG,and NGE were in the regression equation.MAGE ＜3.9 mmol/L,matching rate was 81％,3.9 ≤ MAGE＜7 mmol/L,matching rate was 67％,MAGE≥7 mmol/L,the matching rate was 52％,the difference among 3 groups was statistically significant (P＜0.01).MBG＜7.8 mmol/L,matching rate was 78％,7.8 ≤ MBG＜ 10 mmol/L,matching rate was 68％,MBG ≥10 mmol/L,the matching rate was 57％,the difference among 3 groups was statistically significant (P ＜ 0.01).When Youden index was 0.409 and MAGE was less than 3.745 mmol/L,the match rate was more than 75％.When Youden index was 0.369 and MBG was less than 8.38 mmol/L,the matching rate was more than 75％.The correlation coefficients of all day,fasting,postprandial and nighttime probe values and the corresponding interphalangeal glucose values were 0.958,0.963,0.944,and 0.965 (all P＜0.01).Conclusions The probe values show good consistency with the interphalangeal glucose values,are able to be used for blood glucose assessment.Blood glucose fluctuations influences accuracy of RT-CGM,the smaller the blood glucose fluctuations,the smaller the mean glucose concentration,the smaller the effective blood glucose fluctuations in frequency,the higher would be the accuracy.It seems to be more effective to do calibration during fasting and nighttime.

Objective To investigate the correlation between carotid intima-media thickness (CIMT) and glucose fluctuation in patients with type 2 diabetes mellitus (T2DM).Methods Sixty-eight T2DM patients admitted to Beijing Military General Hospital from March to August 2012 were enrolled in the study,including 32 cases with CIMT thickening (CIMT ≥ 0.9 mm) and 32 cases with normal CIMT (CIMT ＜0.9 mm).The 72 h continuous blood glucose levels were monitored from the day of admission.Results There were no significant differences in the gender,age,body mass index (BMI),systolic and diastolic blood pressure,HDL-C,LDL-C,CHO,TG,glycosylate hemoglobin (HbA1c),fasting blood glucose(FBG) and postprandial average blood glucose (MPBG) between two groups (P ＞ 0.05).The atherosclerosis (AS) score and mean amplitude of glycemic excursion (MAGE),glucose standard deviation,frequency of glycemic excursion (FGE) and absolute mean of daily differences (MODD) in patients with thickening CIMT were (11 ± 7) years,(6.9 ± 3.0) mmol/L,(2.8 ± 1.2) mmol/L,(3.4 ± 1.5) times/d,(2.8 ± 1.3) mmol/L,(4.5 ± 1.0) score,respectively ; while those in patients with normal CIMT were (8 ±6) years,(4.2 ± 1.1) mmol/L,(1.6 ± 0.5) mmol/L,(2.2 ± 0.8) times/d,(2.0 ± 1.0) mmol/L,(3.3 ±0.6) score,respectively.There were significant differences between two groups (all P ＜ 0.05).Pearson correlation showed that CIMT was positively correlated with MAGE,FGE,MODD,course of disease,diastolic blood pressure,LDL-C,MPBG and AS score (P ＜ 0.05).Multiple stepwise regression analysis showed that MAGE,MBPG were the influencing factors of CIMT.Conclusions The CIMT of patients with T2DM is closely correlated with glucose fluctuation,indicating that reduction of blood glucose fluctuation and MAGE,MPBG levels may delay the occurrence of diabetic macroangiopathy.

Objective To investigate the relationshipbetween glucose fluctuation and the degree of nervous dysfunction of the acute cerebral infarction in patients with type 2 diabetes mellitus.Methods 30 patients with ACI and T2DM were chosen as observation group and 30 patients with T2DM without ACI as the control group.Glucose fluctuation in all patients were monitored for 72h with the continuous glucose monitoring system(CGMS).High frequency ultrasound was used to detect the carotid intima-media thickness (IMT).The levels of blood lipids,glycosylated hemoglobin (HbA1c),homocysteinemia(Hcy) and C-reactive protein(C-RP) were detected in all the patients.The national institute of health stroke scale(NIHSS) was performed.The correlation between NIHSS and other observed factors were analyzed.Results (1)The mean amplitude of glycemic excursions (MAGE),blood glucose standard deviation(SDBG),absolute means of daily differences(MODD),the largest amplitude of glycemic excursions(LAGE),blood lipids,HbA1c,Hcy,C-RP and IMT were statistically significant different between the two groups (all P＜0.05); (2)The MAGE,SDBG,IMT,Hey,C-RP,low-density lipoprotein cholesterol-C (LDL-C),and systolic blood pressure(SBP) were correlated with the NIHSS score (all P＜0.05) ;(3)With NIHSS score as the dependent variable and the indicators above as the independent variables,the multiple stepwise regression analysis showed that the MAGE,IMT,Hcy came into the final equation.Conclusions The blood glucose fluctuation is probably the influential factor on the development of acute cerebral infarction in T2DM patients.Therapy for lowering blood glucose smoothly should be established as soon as possible to recover the nerve function after cerebral infarction and reduce the incidence of stroke recurrence.

ObjectiveTo verify the reliability of the mouse model of cerebral cortical microinfarct induced by two-photon microscopy and to explore its pathological changes.MethodsSeventeen male C57BL/6J mice were randomly divided into a microinfarct group (n=11) or a sham operation group (n=6).A thinned cranial window of 3 mm diameter was performed over the cerebral cortex with a high-speed micro-drill until the small blood vessels were clearly observed under a dissecting microscope.Then, a permanent single cortical penetrating arteriole occlusion was induced with a gradually enhanced ultrashort laser irradiation through the thinned cranial window with two-photon microscopy.At 7 days after modeling, the cerebral microinfarct volume was measured with HE staining, and the neuron loss, activation of glial cells and deposition of 3-nitrotyrosine were assessed using immunohistochemistry.ResultsThe target vessels of cerebral cortex in 8 (72.7%) mice were occluded and the microinfarcts formed in the microinfarct group, and the average microinfarct volume was 317.23±20.29 μm3.There were remarkable neuron loss and microglia infiltration in the infarcted core, a large number of reactive astrocytes surrounding the infarcted lesion, and massive deposition of 3-nitrotyrosine in the peri-infarct area.No infarcts were observed in the sham operation group.The deposition of 3-nitrotyrosine in the sham operation group was significantly less than that in the microinfarct group (8.00±1.48 vs.98.38±9.10;t=23.962, P<0.001).Conclusions The mouse model of cerebral cortical microinfarct induced by two-photon microscopy is reliable, and its histopathologic changes are consistent with the pathologic features of cerebral microinfarct.

In order to explore the possibility of applying enzyme histochemistry in biocompatibility evaluation, we investigated the effect of biomaterials on the activities of intracellular enzymes in this experiment. It was found that there was no obvious difference in morphology between osteoblasts co-cultured with HA/TCP and with Ti-alloy. However, transient down-regulation of NADH, SDH, LDH and CCO of the osteoblasts co-cultured with HA/TCP was detected by enzyme histochemistry, but these enzymes of osteoblasts the co-cultured with Ti-alloy were not down-regulated. It was indicated that something extracted from HA/TCP injured the co-cultured osteoblasts slightly. Similar early acute inflammatory reactions were observed after HA/TCP and Ti-alloy were separately implanted into the dorsal muscle of rabbit. There was also no obvious difference between the tissue response to HA/TCP and that to Ti-alloy. Activities of enzymes in tissues around implanted materials were down-regulated at the early injury period and recovered gradually within 30 days post-operation. But the mild toxicity of extracts from HA/TCP was demonstrated by the fact that the recovery period of HA/TCP group was longer than that of Ti-alloy group. It was proved that enzyme histochemistry is more sensitive than tissue morphology analysis in detecting the cell or tissue responses to biomaterials. Therefore, it is possible to use enzyme histochemistry in biocompatibility evaluation.
Alloys ; Animals ; Biocompatible Materials ; chemistry ; Calcium Phosphates ; chemistry ; Cells, Cultured ; Ceramics ; chemistry ; Coculture Techniques ; Female ; Histocytochemistry ; methods ; Hydroxyapatites ; chemistry ; Implants, Experimental ; Male ; Materials Testing ; Osteoblasts ; cytology ; Rabbits ; Titanium

To summarize the clinical experience regarding a patient with early recurrence of atypical hemolytic uremic syndrome (aHUS) after renal transplantation. AHUS is a rare disease with high recurrence rate and poor prognosis. Although the patient was treated with plasma exchange, intravenous gamma globulin, rituximab block B lymphocyte, hormone shock and so on, he still suffered renal transplantation failure. The risk of aHUS recurrence after renal transplantation should be fully evaluated.

Objective To test the effects of TNF-α inhibitor Etanercept on myocardial ischemia/ reperfusion (MI/R) injury in posttraumatic mice,and explore related mechanisms.Methods Traumatic mouse model was established with Noble-Collip drum.Five days after trauma,Notch1 was knocked down by intramyocardial injection of Notch1 small interfering RNAs (siRNA) or scrambled siRNA (20 μg).Seven days after trauma,mice were subjected to MI/R (30 minutes ischemia followed by reperfusion).Sham operation was similarly performed without coronary artery ligation.Ten minutes before reperfusion,mice received Etanercept (8 mg/kg,i.p.).ELISA was used to detect plasma levels of TNF-α and troponin I (cTnI) and myocardial nitrotyrosine content.Twenty-four hours after reperfusion,left ventricular ejection fraction (LVEF) was measured by echocardiography.Infarct size was determined by Evans blue/2,3,5-triphenyl tetrazolium chloride (TTC) double staining.Cardiac caspase-3 activity was detected using a caspase-3 kit.Myocardial TNF-α and Notchl intracellular domains (Notch1 ICD) expressions were determined by Western blot.Chemiluminescence was used to assess myocardial superoxide anion content.Results (1) Compared to vehicle group,Etanercept treatment significantly reduced cTnl content,infarct size and caspase-3 activity (all P ＜0.01),while obviously increased LVEF (P ＜0.01).(2) Etanercept treatment also significantly reduced plasma and myocardial TNF-α contents (P ＜ 0.01),whereas markedly increased myocardial Notch1 ICD content (P ＜0.05).(3) Compared to scrambled siRNA group,Notch1 deficiency significantly increased cTnI content,infarct size and caspase-3 activity (P ＜ 0.05),whereas obviously reduced LVEF (P ＜ 0.05).(4) Etanercept significantly reduced myocardial superoxide anion and nitrotyrosine content (P ＜ 0.01),which was reversed by downregulation of Notch1 (P ＜ 0.05).Conclusions TNF-α inhibitor Etanercept can alleviate MI/R injury after trauma by reducing myocardial oxidative/nitrative stress via activating Notch1 signaling pathway.

Purpose: Pancreatic cancer (PC) is a common malignant tumor of the digestive system, and its 5-year survival rate is only 4%. N6-methyladenosine (m6A) RNA methylation is the most common post-transcriptional modification and dynamically regulates cancer development, while its role in PC treatment remains unclear. Materials and Methods: We treated PC cells with gemcitabine and quantified the overall m6A level with m6A methylation quantification. Real-time quantitative reverse transcription polymerase chain reaction and Western blot analyses were used to detect expression changes of m6A regulators. We verified the m6A modification on the target genes through m6A-immunoprecipitation (IP), and further in vivo experiments and immunofluorescence (IF) assays were applied to verify regulation of gemcitabine on Wilms’ tumor 1–associated protein (WTAP) and MYC. Results: Gemcitabine inhibited the proliferation and migration of PC cells and reduced the overall level of m6A modification. Additionally, the expression of the “writer” WTAP was significantly downregulated after gemcitabine treatment. We knocked down WTAP in cells and found target gene MYC expression was significantly downregulated, m6A-IP also confirmed the m6A modification on MYC. Our experiments showed that m6A-MYC may be recognized by the “reader” IGF2BP1. In vivo experiments revealed gemcitabine inhibited the tumorigenic ability of PC cells. IF analysis also showed that gemcitabine inhibited the expression of WTAP and MYC, which displayed a significant trend of co-expression. Conclusion Our study confirmed that gemcitabine interferes with WTAP protein expression in PC, reduces m6A modification on MYC and RNA stability, thereby inhibiting the downstream pathway of MYC, and inhibits the progression of PC.

Objective:To explore the risk factors of paralytic ileus (PI) after simultaneous pancreas-kidney (SPK) transplantation.Methods:From January 2017 to December 2019, clinical data were reviewed retrospectively for 115 cases of SPK transplantation. The risk factors of PI after SPK were analyzed. According to the occurrence of PI, they were divided into two groups of occurrence and non-occurrence. One-way analysis of variance was utilized for analyzing such influencing factors as gender, age, body mass index (BMI), diabetic type, duration of diabetes, mode of dialysis, duration of dialysis, diabetic gastroenterology, history of open surgery, bowel preparation, operative duration, hemorrhagic volume, immunosuppressant and hypoproteinemia. Multivariate Logistic regression analysis was performed for screening the suspected risk factors.Results:Among them, 19 patients (16.5%) had PI. Univariate analysis showed that PI was associated with diabetic gastroenterology, operative duration, history of open surgery, no bowel preparation and hypoproteinemia ( P<0.05). Multivariate Logistic regression analysis revealed that the risk factors of PI after SPK included diabetic gastroenterology, operative duration time, history of open surgery and no bowel preparation ( P<0.05). Conclusions:Diabetic gastroenterology, operative duration, history of open surgery and no bowel preparation are risk factors for PI after SPK. Clinical interventions for the above factors are necessary.