Taxol is one of the most potent anti-cancer agents, which is extracted from the plants of Taxus species. Isopentenyl diphosphate isomerase (IPI) catalyzes the reversible transformation between IPP and DMAPP, both of which are the general 5-carbon precursors for taxol biosynthesis. In the present study, a new gene encoding IPI was cloned from Taxus media (namely TmIPI with the GenBank Accession Number KP970677) for the first time. The full-length cDNA of TmIPI was 1 232 bps encoding a polypeptide with 233 amino acids, in which the conserved domain Nudix was found. Bioinformatic analysis indicated that the sequence of TmIPI was highly similar to those of other plant IPI proteins, and the phylogenetic analysis showed that there were two clades of plant IPI proteins, including IPIs of angiosperm plants and IPIs of gymnosperm plants. TmIPI belonged to the clade of gymnosperm plant IPIs, and this was consistent with the fact that Taxus media is a plant species of gymnosperm. Southern blotting analysis demonstrated that there was a gene family of IPI in Taxus media. Finally, functional verification was applied to identify the function of TmIPI. The results showed that biosynthesis of β-carotenoid was enhanced by overexpressing TmIPI in the engineered E. coli strain, and this suggested that TmIPI might be a key gene involved in isoprenoid/terpenoid biosynthesis.

Objective:To explore the etiology and characteristics of chronic wet cough in children in Qingdao.Methods:Patients with chronic wet cough treated at respiratory clinic of the Women and Children′s Hospital Affiliated to Qingdao University from July 2018 to June 2019 were included in this study.After three-month follow-up, the etiological data was analyzed.Results:(1)A total of 213 children were included, ranging in age from 1 month to 14 years old, including 38 cases of 1 month~1 year old, 47 cases of 1~3 years old, 87 cases of 3~6 years old, and 41 cases of 6~14 years old.The median age was 4.7 years.The top four causes of chronic wet cough in children were upper airway cough syndrome(33.8%), protracted bacterial bronchitis(20.7%), asthma with upper airway cough syndrome(15.5%), and asthma with infection(10.8%). Other causes were postinfection cough, pertussis syndrome, bronchiectasis, gastroesophageal reflux, bronchial foreign body, abnormal airway development, cystic fibrosis and so on.(2)The first cause of chronic wet cough in different age groups: 1 month to 3 years old group was protracted bacterial bronchitis; 3 to 14 years old group was upper airway cough syndrome.(3)The causes of chronic wet cough showed seasonal differences.Upper airway cough syndrome and cough after infection had a more balanced incidence throughout the year; protracted bacterial bronchitis and pertussis syndrome were common in winter; asthma with upper airway cough syndrome and asthma with infection were common in spring and autumn.Conclusion:Upper airway cough syndrome, protracted bacterial bronchitis, asthma with upper airway cough syndrome, and asthma with infection are the 4 leading causes for children with chronic wet cough in Qingdao.The causes of chronic wet cough have age and seasonal differences.

Objective To observe the effect of methylprednisolone on the expression levels of aquaporin-4 (AQP-4) and S100 protein in brain tissue of hypertensive intracerebral hemorrhage in rats and to investigate the treatment timing and the possible neuroprotective mechanism of methylprednisolone for hypertensive intracerebral hemorrhage. Methods Sixty-four rats were randomly divided into 3 groups:a sham operation group ( n= 8 ), a control group ( n= 8 ), and a methylprednisolone group ( n= 48 ). The methylprednisolone group was redivided into 2, 4, 8, 12, 24, and 48 hsubgroups (n=8 in each subgroup) according to the modeling to the time intervals of methylprednisolone treatment. Methylprednisolone was administered intraperitoneal y (30 mg/kg fol owed by 15 mg/kg every 6 hours for 3 d) at the corresponding time points in the methylprednisolone group, and the equal volume normal saline was administered intraperitoneal y in the control group. Neurological behavior score was conducted at 24 and 72 h after methylprednisolone treatment. The dry-wet weight method was used to measure hemispheric water content. In situ hybridization and immunohistochemical staining were used to detect the expression changes of AQP-4 mRNA and AQP-4 protein respectively. Double staining immunohistochemistry was used to detect AQP-4 and S100 protein. Results Compared with the sham operation group, the expression level of AQP-4 and brain water content in the control group were significantly increased (al P<0. 05). Compared with the control group, the neurological scores, expression levels of AQP-4 mRNA and protein, as wel as the brain water content in early methylprednisolone subgroups (2 h, 4 h and 8 hsubgroups) were significantly decreased (al P<0. 05). Double staining immunohistochemistry showed that expression levels of AQP-4 and S100 protein in early methylprednisolone subgroups (2 h, 4 h and 8 hsubgroups) were significantly decreased than those in the control group (al P<0. 05). Conclusions Early methylprednisolone may downregulate the expression levels of AQP-4 and S100 protein in the brain tissue after hypertensive intracerebral hemorrhage in rats, and thus attenuate brain edema after intracerebral hemorrhage.

AIM: To explore the histocompatibility of pinealocyte microencapsules in vivo. METHODS: The pineal glands of neonatal rats were removed under operating microscope and pinealocytes were isolated through collagenase and trypsin digestion. Pinealocytes were cultured for one week in vitro and collected immediately after digesting was encapsulated in APA microencapsules. The cells and empty capsules were transplanted into abdominal cavity or intermuscular space respectively and retrieved at the 15th or 30th day after operation. Morphological observation, HE staining, cell counting, and HPLC technique were used to analyze the shape, proliferation and function, the degree of inflammation fibrosis of retrieved microencapsules. RESULTS: The retrieve rate of cell capsule from abdominal cavity was about 85 % . The retrieved capsules had integrated profile mostly although some were damaged. The amount of macrophages attached to capsule wall and the thickness of wall increased gradually following the period of transplantation. However, the retrieve rate, wall thickness had no difference between retrieved cell and empty capsules at the same time. Secretion ability of pinealocytes in capsule retrieved at 15th day after operation decreased rapidly and those retrieved at 30th day after operation lossed secretion function. CONCLUSION: APA microencapsules had histocompatibility relatively in vivo and protected pinealocytes in capsule from immunologic rejection of the host. The survival time was about 20 days. During this period cells in capsule maintained activity and MT secretion ability.

ObjectiveTo investigate the molecular mechanism of prostaglandins E2 ( PGE2 ) in promoting bone formation by detecting the changes of gene expression profiles of MC3T3-E1 osteoblasts treated with PGE2. MethodsThe genes with differential expression in MC3T3-E1 osteoblasts treated with 10 μmol/L PGE2 for 30 minutes were performed by gene chip technology. Several major genes during bone regeneration were selected for Western blot analysis. ResultsAfter co-culture of MC3T3-E1 cells with PGE2 at concentration of 10 μmol/L for 30 min, 276 genes were up-regulated, including bone regeneration related MMD (monocyte to macrophage differentiation associated), NR4A2 (nuclear receptor subfamily 4, group A, member 2), BMP-7 ( bone morphogenetic protein-7), POSTN ( periostin, osteoblast specific factor) and catenin (cadherin-associated protein) genes; and 168 genes were down-regulated,including bone regeneration related Idl ,2,3 ( inhibitor of DNA binding 1,2,3 ) genes. Western blot analysis indicated that the expressions of nuclear factor (NF)-κB p65 and BMP-7 protein in the osteoblasts treated with 10 μmol/l PGE2 were apparently higher ( P ＜ 0. Ol ) than that of the controls, whereas the ld2 expression decreased (P ＜0. O1 ) under the same conditions, which was almost the same as the results of gene chip technology. ConclusionsWith the results of gene chip and Western blot, it can be speculated that the PGE 2 firstly activates the nuclear receptor NR4A2 and then the nuclear transcription factor NF-κB, induces the changes of the downstream gene BMP-7 and Id2 expression and finally results in the differentiation of the osteoblasts and promote the bone regeneration.

Objective To study the effects of Schwann cell-derived neurotrophic factor(SDNF) on myoblast stem cells(called satellite cells,SCs) in vitro. Methods After setting up SCs culture system in vitro, SCs which treated with various SDNF concentrations culture medium were dynamically evaluated by cell morphology,MTT growth curve and fusion rate. Results The ability of SCs preceding their participation in muscle repair include proliferation and differentiation, 200 ng/ml SDNF stimulated cell proliferation more than the other medium,but 50 ng/ml,100 ng/ml,200 ng/ml,400 ng/ml SDNF made SCs differentiation significantly for their high myotube fusion rate. Conclusion SDNF can regulate the proliferation and differentiation of rat skeletal satellite cellsin vitro,but in differentiation significantly.SDNF might play a role in slowing down denervated muscle atrophy.

Objective The aim of the present study was to evaluate the efficacy of three different modalities in treatment of lung oligometastases from nasopharyngeal carcinoma ( NPC) after radiotherapy and to identify a more appropriate treatment modality.Methods The clinical data of 87 cases of lung oligometastases from NPC were analyzed retrospectively.Among them, 33 patients underwent local small-field irradiation+/-chemotherapy, 28 underwent whole-lung irradiation+chemotherapy, and 26 underwent simple chemotherapy.The survival rates were calculated using Kaplan-Meier analysis.The differences among the modalities were evaluated using the log-rank test.Cox univariate and multivariate analyses were performed to determine the influencing factors.Results The 3-year lung metastasis survival ( LMS) rates of patients with lung metastasis undergoing the three treatment modalities ( local small-field irradiation +/-chemotherapy, whole-lung irradiation+chemotherapy and chemotherapy alone) were 89.3%, 72.7%, and 72.4%, respectively, showing a significant difference between the groups ( P=0.003 ) .Further subgroup analysis showed that the 5-year LMS rate was significantly higher in the local small-field irradiation+/-chemotherapy group than that in the whole-lung irradiation+chemotherapy group and chemotherapy alone group (P=0.001).The 2y-ear progression -free survival (PFS ) rates of the three groups were 571.%, 25.8%and 3.8%, respectively, showing significant intergroup differences ( P=0.002 and P<0 .001 ) . Multivariate analysis indicated that compared with the whole lung irradiation group and the chemotherapy alone group, the local irradiation+/-chemotherapy is an independent favorable prognostic factor for LMS and PFS (P<0.05). Conclusion Local radiotherapy combined with systemic chemotherapy is the best therapeutic modality for lung oligometastases derived from NPC after radiotherapy, improving the LMS and prolonging the PFS.

Objective The aim of the present study was to evaluate the efficacy of three different modalities in treatment of lung oligometastases from nasopharyngeal carcinoma ( NPC) after radiotherapy and to identify a more appropriate treatment modality.Methods The clinical data of 87 cases of lung oligometastases from NPC were analyzed retrospectively.Among them, 33 patients underwent local small-field irradiation+/-chemotherapy, 28 underwent whole-lung irradiation+chemotherapy, and 26 underwent simple chemotherapy.The survival rates were calculated using Kaplan-Meier analysis.The differences among the modalities were evaluated using the log-rank test.Cox univariate and multivariate analyses were performed to determine the influencing factors.Results The 3-year lung metastasis survival ( LMS) rates of patients with lung metastasis undergoing the three treatment modalities ( local small-field irradiation +/-chemotherapy, whole-lung irradiation+chemotherapy and chemotherapy alone) were 89.3%, 72.7%, and 72.4%, respectively, showing a significant difference between the groups ( P=0.003 ) .Further subgroup analysis showed that the 5-year LMS rate was significantly higher in the local small-field irradiation+/-chemotherapy group than that in the whole-lung irradiation+chemotherapy group and chemotherapy alone group (P=0.001).The 2y-ear progression -free survival (PFS ) rates of the three groups were 571.%, 25.8%and 3.8%, respectively, showing significant intergroup differences ( P=0.002 and P<0 .001 ) . Multivariate analysis indicated that compared with the whole lung irradiation group and the chemotherapy alone group, the local irradiation+/-chemotherapy is an independent favorable prognostic factor for LMS and PFS (P<0.05). Conclusion Local radiotherapy combined with systemic chemotherapy is the best therapeutic modality for lung oligometastases derived from NPC after radiotherapy, improving the LMS and prolonging the PFS.

Objective To investigate the effects of fasting serum triglycerides (TG) levels at different baseline on the risk of new-onset acute pancreatitis (AP) in in-service and retired employees of Kailuan Group.Methods A total of 125 178 in-service and retired employees of Kailuan Group who received health check-ups from 2006 to 2009 and had no AP history but had complete TG data were prospectively enrolled.According to quantile level,the baseline serum fasting TG level of study subjects were divided into ＜1.01 mmol/L group (n=42 128),1.01 to 1.64 mmol/L group (n=41 711) and ＞ 1.64 mmol/L group (n=41 339).The incidence of new-onset AP of these three groups was analyzed.The survival curve was plotted by Kaplan-Meier method.The cumulative incidence rate was calculated and tested by log-rank method.And multivariate Cox proportional hazards regression model was performed to calculate hazard ratios (HR) of baseline fasting serum TG level for AP.Results After followed up for (7.36±1.23) years,a total of 193 cases of AP occurred.The incidences of AP in ＜1.01 mmol/L group,1.01 to 1.64 mmol/L group and ＞ 1.64 mmol/L group were 1.43 events/10 000 person-years,2.37 events/10 000 person-years and 2.49 events/10 000 person-years,respectively.The cumulative incidence rates of AP in ＜1.01 mmol/L group,1.01 to 1.64 mmol/L group and ＞1.64 mmol/L group were 0.10％ (44/42 128),0.18％ (73/41 711) and 0.18％ (76/41 339),respectively,and the difference was statistically significant (x2 =9.998,P=0.007).The results of multivariate Cox proportional hazards regression model analysis indicated that the risk of AP increased in 1.01 to 1.64 mmol/L group and ＞ 1.64 mmol/L group compared with that of ＜1.01 mmol/L group,HR and 95％ confidence interval (CI) were 1.56 (1.07 to 2.29) and 1.57 (1.06 to 2.32),respectively.After excluded onset AP within one year,with a control group of ＜1.01 mmol/L group,the results of multivariate Cox proportional hazards regression model analysis indicated that the HR and 95％CI for AP of 1.01 to 1.64 mmol/L group and ＞ 1.64 mmol/L group were 1.70 (1.11 to 2.58) and 1.69 (1.10 to 2.60),respectively.Conclusion Baseline fasting serum TG levels over 1.01 mmol/L may increase the risk of AP.

Objective:To investigate the effects of body mass index (BMI) levels at different baseline on the risk of new-onset acute pancreatitis (AP).Methods:The subjects were from the Kailuan Study Cohort and divided into 3 groups according to baseline BMI levels: BMI<24 kg/m 2, normal weight; BMI 24-28 kg/m 2, overweight; BMI≥28 kg/m 2, obesity. The incidence of new-onset AP in these three groups was analyzed. The survival curve was plotted by Kaplan-Meier method, the cumulative incidence was calculated and tested by log-rank method. Multivariate Cox proportional hazards regression model was used to calculate HR of baseline BMI levels for AP. Results:A total of 123 841 subjects were included and followed up for (11.94±2.13) years, during which, 395 cases were found with AP. The incidence of AP was 2.67 per 10 000 person years in total population, and the incidences of AP were 2.20, 2.72 and 3.58 per 10 000 person-years in the normal, overweight and obesity groups, respectively. The cumulative incidences of AP was 0.32%, 0.40% and 0.49% in normal, overweight and obesity groups, respectively, which showed a significant inter-group difference by log-rank test ( χ2=13.17, P<0.01). The results of multivariable adjusted Cox proportional hazards regression model analysis indicated that obesity group ( HR=1.45, 95% CI: 1.10-1.92) had a higher risk for AP compared with the normal BMI group. The subgroup analyses by age and sex showed that compared with the normal weight group,the HRs for AP in the obesity group was 1.58(95% CI:1.14-2.19) and 1.40(95% CI:1.03-1.90) among subjects younger than 60 years old and male subjects, respectively. After excluded onset AP within two years from baseline,with a control group from normal weight,the results of multivariate Cox proportional hazards regression model analysis indicated that the AP in the obesity group was 1.60 (95% CI: 1.18-2.15). Conclusion:Obesity may increase the risk of developing AP, particularly among young and middle-aged men.