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.

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.

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.

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.

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 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 explore the impact of oral nutritional supplements in the local advanced nasopharyngeal carcinoma patients receiving chemoradiotherapy. Methods From 2016 to 2018, we prospectively collected 114 patients into our study, including intervention group ( n=58) and control group ( n=56) . Patients in the intervention group were given oral nutritional supplements from the beginning of radiotherapy, while the control group patients were receiving usual diet care. We collected data of weight, hematological index and nutrition related information before, during and after treatments. Results All patients experienced weight loss during radiotherapy. The declining trend was more obvious in intervention group but with no statistical difference ( P>0.05) . Radiotherapy interruption was lower, completion rate of 2 cycles of concurrent chemotherapy was higher( 78% vs 64%, P=0.02) and level of total serum protein and albumin was more stable ( P=0.003, 0.001) in the intervention group. No difference was found in acute toxicities between the two groups ( P>0.05) . During the treatment period, the nutrition risk screening score declined, more patients with NRS 2002 ≥3 in the control group than the intervention group ( P<0.05) , while no statistically significant difference was found in PG-SGA between the two groups ( P>0.05 ) . Conclusions Oral nutritional supplements can improve treatment tolerance and keep the stability of serum protein level, nevertheless, it has no obvious advantage in maintenance of weight and short-term nutritional status

Ankylosing spondylitis (AS) combined with spinal fractures with thoracic and lumbar fracture as the most common type shows characteristics of unstable fracture, high incidence of nerve injury, high mortality and high disability rate. The diagnosis may be missed because it is mostly caused by low-energy injury, when spinal rigidity and osteoporosis have a great impact on the accuracy of imaging examination. At the same time, the treatment choices are controversial, with no relevant specifications. Non-operative treatments can easily lead to bone nonunion, pseudoarthrosis and delayed nerve injury, while surgeries may be failed due to internal fixation failure. At present, there are no evidence-based guidelines for the diagnosis and treatment of AS combined with thoracic and lumbar fracture. In this context, the Spinal Trauma Academic Group of Orthopedics Branch of Chinese Medical Doctor Association organized experts to formulate the Clinical guideline for the diagnosis and treatment of adult ankylosing spondylitis combined with thoracolumbar fracture ( version 2023) by following the principles of evidence-based medicine and systematically review related literatures. Ten recommendations on the diagnosis, imaging evaluation, classification and treatment of AS combined with thoracic and lumbar fracture were put forward, aiming to standardize the clinical diagnosis and treatment of such disorder.