Aim of the present study is to investigate activation effect of nobiletin on cystic fibrosis transmembrane conductance regulator (CFTR) chloride channel activity. CFTR-mediated iodide influx assay and patch-clamp tests were done on FRT cells stably co-transfected with human CFTR and EYFP/H148Q. Nobiletin potently activated CFTR chloride channel activity in a dose- and time-dependent manner. The CFTR blocker CFTR(inh)-172 could completely reverse the effect. Preliminary mechanism study indicated that nobiletin activated CFTR chloride channel through a direct binding way. In addition, ex vivo tests done on mice trachea showed that nobiletin time-dependently stimulated submucosal gland fluid secretion. Nobiletin may be a therapeutic lead compound in treating CFTR-related diseases including disseminated bronchiectasis.

BACKGROUND: Tiantai I consists of gastrodia, Chinese angelica root, areca seed. It has been considered as the roles of invigorating the liver, nourishing marrow, heightening the intelligence and causing resuscitation. OBJECTIVE: To observe the effects of Tiantai Ⅰ on the abilities of learn ing and memory and the activity of central cholinergic system in mice withspontaneous Alzheimer disease. DESIGN: A randomized control observation. SETTING: Shenzhen Institute of Integrated Chinese and Western Medicine. MATERIALS: Male Kunming mice of 13 months old were raised to 21 months old, of which 52 with spontaneous Alzheimer disease werescreened. They were randomly divided into blank control group, westerndrug control group, Tiantai Ⅰ 6.80 and 20.41 g/kg groups, 13 mice in eachgroup. Another 13 aged mice with normal learning and memory abilitieswere selected as the normal control group at the same time. METHODS: Mice in the western drug control group were treated with0.6 mg/Kg Hydergine, those in the Tiantai Ⅰ 6.80 and 20.41 g/kg groupswere given intragastric administration of Tiantai Ⅰ of 6.80 and 20.41 g/kg, respectively, and those in the normal control group and blank control groupwere given double distilled water of the same volume. The learning andmemory results were examined by the step-down test. Freezing sections ofbrain tissue were prepared, acetylcholinesterase (AChE) fiber was showedaccording to the Hedreen method, and choline acetyltransferase (ChAT) was detected with Burt and Silver methods, the automatic image analysissystem for biomedical application was applied in the quantitative analysisof AChE fiber and ChAT activity. MAIN OUTCOME MEASURES: ① Effect of Tiantai Ⅰ on the abilities of learning and memory in mice with Alzheimer disease; ② AChE fiberarea density in temporal cortex and hippocampal CA1 region; ③ ChAT ac tivity in Meynert nuclei of basal forebrain. RESULTS: ① Tiantai Ⅰ in ameliorating the abilities of learning and mem ory in mice with spontaneous Alzheimer disease: The error times in theTiantai Ⅰ 6.80 and 20.41 g/kg groups, normal control group and westerndrug control group were obviously fewer than that in the blank controlgroup (5.08±2.18, 3.77±1.48, 3.25±1.27, 6.17±3.12, 9.31±3.65, P ＜ 0.05). ② AChE fiber area density: The AChE fiber area densities in temporalcortex were obviously higher in the Tiantai Ⅰ 6.80 and 20.41 g/kg groups, normal control group and western drug control group than in the blankcontrol group (20.19±4.18, 25.01±6.44, 26.38±6.52, 19.44±4.33, 14.36±3.21, P ＜ 0.05). The AChE fiber area densities in hippocampal CA1 region were obviously higher in the Tiantai Ⅰ 20.41 g/kg groups and nor mal control group than in the blank control group, western drug controlgroup and Tiantai Ⅰ 6.80 g/kg group (13.88±3.07, 14.11±4.36, 6.57±2.30, 10.13±2.94, 9.63±2.40, P ＜ 0.05). ③ ChAT activity: It was obviously high er in the TiantaiI 20.41 and 6.80 g/kg groups, western drug control groupand normal control group than in the blank control group (0.683±0.179, 0.589±0.179, 0.552±0.173, 0.705±0.141, 0.354±0.131, P ＜ 0.05). CONCLUSION: Tiantai Ⅰ can significantly improve the disturbances oflearning and memory in mice with spontaneous Alzheimer disease, and itcan increase the activity of central cholinergic system in a dose-dependentmanner.

BACKGROUND: The theory of free radical on onset of Alzheimer disease(AD) and the relationship between oxidative damage and onset of AD have made anti-oxidative drugs become the hot topic in researches. Whether does the action of natural Chinese herbs on anti-injury of oxygenic free radical produce intervention on AD?OBJECTIVE: To observe the effects of developed and researched tiantai I,the natural Chinese herb, on learning and memory function and metabolism of oxygenic free radical in the model of spontaneous AD.DESIGN: Completely randomized experimental study based on the experimental animals.SETTING: Clinical Institute of Chinese-Western integration of City-grade.MATERIALS: The experiment was performed in 2nd grade Animal Experimental Room of Clinical Institute of Chinese-Western integration in Shenzhen. Kunming male mice of clean grade were employed, at the age of 13 months, mass weighted varied from 40 to 50 g and from Experimental Animal Center of Zhongshan Medical University(Qualified No. 95A07).INTERVENTIONS: When the experimental animals were bred till 21 months, AD mice were screened from the aged animals according to International Universal Standards and Methods. The experimental animals were randomized into blank control, western-drug control, the small dosage of tiantai I group(small dosage group), the large dosage of tiantai I group(large dosage group) and learning and senile normal-memory group(senile normal group) in addition. In western-drug control, hydergine 0.6 mg/kg was administrated, and in small and large groups, tiantai I of 6. 80 g/kg and 20.41 g/kg were administrated respectively, continuously for 60 days. The double-distilled water of equal dosage was infused in both blank control and senile normal group.MAIN OUTCOME MEASURES: Step-down test was applied to determine learning and memory scores. Froze sections of brain tissue were prepared and immuno-histological-chemical method was used to display the superoxidative dismutase(SOD) activity in cerebral cortex and hippocampus. Ultrathin section of brain tissue in hippocampal CA1 was prepared and transmission electron microscope(TEM) was used to observe cellular lipofuscin in neuron. Automatic image analysis system for biomedical application was used to determine in quantity SOD and lipofuscin.RESULTS: Learning and memory of AD mice were significantly decreased ( P ＜ 0.01 ) . Tiantai I improved significantly learning and memory disturbance of mice in the model of spontaneous AD( P ＜ 0.05- 0.01),markedly enhanced SOD activity in central cerebral cortex and hippocampus ( P ＜ 0.05 - 0.01 ) and reduced cellular lipofuscin content of neuron( P＜ 0.05 -0.01 ). In addition, it provided certain of quantitative reaction.CONCLUSION: For the mice with spontaneous senile dementia, learning and memory as well as SOD activity of neuron in relevant cerebral areas are decreased remarkably and lipofuscin is increased significantly. Tiantai I acts significantly in pharmacodynamics on anti-injury of oxygenic free radical,increases significantly SOD activity, decreases lipofuscin content and improves the ultrastructure of neuron in cortex and hippocampus in AD mice.

BACKGROUND: The synaptic plasticity of central nerves is the key basis of behavior-dependent learning and memory, there have been few reports about weather the Chinese herbal medicine for treatment of Alzheimer disease is through enhancing synaptic plasticity to improve learning and memory.OBJECTIVE: To investigate the effect of tiantai No. 1 recipe on learning and memory function and synaptic plasticity of spontaneous Alzheimer disease models.DESIGN: A randomized and controlled trial.SETTING: Institute of Combination of Traditional Chinese Medicine and Western Medicine of Shenzhen.MATERIALS: The experiment was completed at the grade Ⅱ Laboratory of Institute of Combination of Traditional Chinese Medicine and Western Medicine of Shenzhen City. The experimental animals were mice of Kunming strain.METHODS: Totally 52 Kunming mice, aged 21 months, with spontaneous Alzheimer disease (memory impairment) were chosen, and at random divided into 4 groups: senile dementia, western drug control, tiantai No. 1recipe 6.80 g/kg and 20.41 g/kg groups; besides 13 mice with normal learning and memory power were chosen as normal senile group. Hydergine 0.6 mg/kg was given for mice in western drug control group, tiantai No. 1recipe 6.80 g/kg and 20.41 g/kg were respectively given for mice in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups. The medicines above were all prepared into 0.5 mL solution and given by gavage, for consecutive 60 days. The mice in normal senile and senile dementia groups were given double distilled water of the same volume. The learning memory power was tested by jumping stand; ultrathin cerebral section of hippocampal CA1 region was examined with transmission electron microscope,and some parameters of synaptic plasticity was also quantitatively detected with automatic micrographic analysis system.MAIN OUTCOME MEASURES: ① The effect of tiantai No. 1 recipe in different doses on learning and memory of spontaneous Alzheimer disease mice. ② Observation of synapse with electron microscope and quantitative examination stereologically.RESULTS: All animals involved in the trial entered the final result analysis without any loss. ① The effect of tiantai No. 1 recipe in different doses on learning and memory of spontaneous Alzheimer disease mice: The error numbers of learning and memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were smaller than that in senile dementia group,and the error number of learning in tiantai No. 1 recipe 6.80 g/kg group was smaller than that in tiantai No. 1 recipe 20.41 g/kg group. The escape latent periods of learning training was smaller than that in senile dementia group and the safe platform latent periods of memory in tiantai No. 1 recipe 6.80 g/kg and 20.41 g/kg groups were longer than that in senile dementia group, and the safe platform latent period of memory in tiantai No. 1 recipe 6.80 g/kg group was smaller than that in 20.41 g/kg group. ② Observation of synapse with electron microscope and quantitative examination stereologically: Compared with the mice in normal senile group, for mice in senile dementia group, there occurred degenerative phenomena: the number of synapses reduced, a part of synaptic clefts were not clear, the synaptic junction was interrupted, and he sizes of synaptic vesicles were different; in the rest groups there were also degenerative phenomena, however the degrees were milder than those in the model group. Tiantai No. 1 recipe could remarkably raise the number density and sectional density of synapses in the hippocampal CA1 region, and the increased extents in 20.41 g/kg tiantai No.1 recipe was larger than those in 6.80 g/kg tiantai No. 1 recipe.CONCLUSION: Tiantai No. 1 recipe can remarkably improve their learning and memory impairments, which may be related with that it promotes re-generation of synapse and improves the plasticity of cerebral synapses;and the action presented a relationship of dose-effect.

Objective To explore the relationship between T-cell acute lymphoblastic leukemia and the Notch signaling pathway.Methods Human T-cell acute lymphoblastic leukemia SupT1 cells were infected with the lentiviral vector made up specific Notch1-shRNA gene and nonspecific Notch1-shRNA gene.The inhibitive rate of SupT1 cells was detected by CCK-8.The rates of early apoptotic cells (Annexin V+/ 7-AAD-) and late apoptotic cells (Annexin V+/7-AAD+) were analyzed by flow cytometry and the expression levels of Notch1 receptor gene and downstream target genes were assessed by quantitative reverse transcription and polymerase chain reaction (QT-PCR).Results The cell inhibition rates of Notch1 interference group,control group and empty vector group at 96 h were 0.902±0.013,0,and 0.486±0.084,respectively,and it was increased obviously in Notch1 interference group (both P ＜ 0.05).The cell early apoptosis rates of the three groups were (15.27±0.31) ％,(5.57±0.25) ％,(5.80±0.20) ％,respectively.The cell early apoptosis rate of Notch1 interference group was increased obviously compared with the control group and empty vector group (both P ＜ 0.05).While the cell late apoptosis rates had no significant difference among the three groups (P ＞ 0.05).The mRNA expression levels of Notch1 receptor gene and its target genes (Hesl,c-myc,NF-κB) at 48 h,72 h and 96 h were higher than those in the control group and empty vector group (all P ＜ 0.05).Conclusions The specificity of Notch1-shRNA can effectively decrease the Notch1 mRNA expression,and reduce the expression level of downstream target genes.Notch1 cut can inhibit the proliferation of SupT1 cells,and promote the early apoptosis.

Objective To analyze the clinical features and risk factors for an emerging infection during the first induction chemotherapy in elderly patients with acute leukemia.Methods A retrospective analysis of clinical data of 79 elderly patients with newly diagnosed acute leukemia was performed in Second Hospital of Shanxi Medical University from January 2014 to May 2016.Results The 70 cases among 79 elderly patients with acute leukemia were suffered from infection with infection incidence rate of 88.6％ (70/79)during first induction chemotherapy.The infection-related fatality rate was 8.6 ％ (6/70).Being clear about sites of infection accounted for 90.0 ％ (63/70),and the top three infection sites were the lungs,gastrointestinal tract and the bloodstream.113 pathogenic strains were detected,including gram-negative bacilli accounting for 42.5 ％ (48/113),Gram-positive cocci for 30.1％ (34/113),fungi for 24.8％ (28/113),the virus for 2.7％ (3/113).Based on clinically and confirmatively diagnosis,the invasive fungal diseases mostly as Candida accounted for 30.4 ％ (24/79),mixed infections accounted for 34.3％ (24/70).Univariate analysis showed agranulocytosis and AML were risk factors for infection.Logistic multivariate regression analysis showed that agranulocytosis was a risk factor for infection (OR=12.010,95％CI:2.346-107.973,P=0.000).The infection does not affect a complete remission rate of acute leukemia (x2 =0.001,P=0.983).Conclusions For newly diagnosed elderly acute leukemia patients,an emerging infection during the first induction chemotherapy is characterized by a high incidence,high fungal infection rate,most common site in lung,Gram-negative bacteria as most common pathogen,and an increased infection rate by agranulocytosis.The infection does not affect the remission rate of acute leukemia.

Objective To investigate the relevance between CD25 and common markers of immune phenotype in acute myeloid leukemia (AML), and the relationship between CD25 and FLT3-ITD gene. Methods The expression of mono-clonal antibodies, including, CD7, CD117, CD33, CD34, CD38, cyMPO, CD13, CD36, CD56, CD11b, CD19, CD16, CD14, CD64 and interleukin-2 three receptor chain CD25 (αchain), CD122 (β chain), CD132 (γ chain) were detected by immunofluorescence flow cytometry in 36 cases of AML bone marrow blast. The expression of every group of pa-tients with FLT3-ITD gene were detected by PCR. Results In 36 cases of AML patients, the rate of CD25+was 13.89%(5/36), 12.5% (1/8) in AML-M2 group, 18.18% (2/11) in AML-M4 group, 20% (2/10) in AML-M5 group, respectively, and expression rate of CD25+had been no statistically differences between those subgroups (P> 0.05). The percentage of bone marrow blasts which express CD7, CD117, CD33, CD34, CD38, cyMPO, CD13, CD36, CD56, CD11b, CD19, CD16, CD14, CD64, CD122, CD132 monoclonal antibodies has no significant difference between CD25+AML-M2 group and CD25-AML-M2 group, CD25+AML-M4 group and CD25-AML-M4 group, CD25+AML-M5 group and CD25-AML-M5 group, respectively (P> 0.05). Seven cases of AML patients with FLT3-ITD mutation, in five cases of CD25+AML pa-tients three case accompany FLT3-ITD+ mutation, the rates of FLT3-ITD+in CD25+AML was 60%, and higher than CD25-AML group (P <0.05). Conclusion (1)The expression rate of CD25 in AML patients is lower, and no significant correlation in patients in different AML FAB subtypes and common myeloid phenotype markers. (2)Since the FLT3-ITD mutation rates are higher in CD25+AML patients, there is important significance to clear the correlation between CD25 and FLT3-ITD gene.

Objective To examine the influencing factors related to clinical efficacy and outcomes of adult primary immune thrombocytopenia (ITP).Methods The clinical data of 161 cases of ITP admitted in the Second Hospital of Shanxi Medical University from June 2013 to March 2017 were collected.The influencing factors related to clinical efficacy and prognosis of adult ITP patients were analyzed.Results There were 60 males and 101 females with a M/F ratio of 0.59∶1 and a median age of 45 years (18-84 years).There were 109 newly diagnosed ITP cases,14 persistent ITP cases and 38 chronic ITP cases in this series.Seventy nine patients received intravenous immunoglobulin g (IVIg) treatment and 82 patients received high dose-dexamethasone treatment.There were no significant differences in clinical efficacy [91.13％(72/79) vs.87.80％(72/82),x2=0.181,P=0.914] and relapse rate [36.11％(26/72) vs.30.55％(22/72),x2=0.189,P=0.910] between IVIg and high dose-dexamethosone groups.Multivariate regression analysis showed that bleeding score ≥2 was the independent risk factor for the lower clinical efficacy (RR=1.415,95％CI:1.008-1.986,P＜0.05).Patients were followed up for a median of 9.0 months (0.5-55.0 months),48 patients relapsed with a relapse rate of 33.33％ and a median relapse time of 1.8 months (0.5-24.0 months).Conclusions IVIg and high dose-dexamethasone have the similar clinical efficacy and relapse rate for treatment of adult ITP.The patients with the bleeding score ≥2 are more likely to get lower remission rate.

Objective:To summarize the clinical manifestations, treatment and outcome of neonatal pseudo-Bartter syndrome caused by maternal hyperemesis gravidarum.Methods:This retrospective study collected the clinical data of a set of premature twins with pseudo-Bartter syndrome who were admitted to Hebei Children's Hospital in September 2022. Clinical features of the cases were summarized with descriptive analysis.Results:The twins born with a gestational age of 30 +3 weeks required tracheal intubation and mechanical ventilation due to premature birth and respiratory distress. They were transferred to our hospital 2 h after birth. The mother suffered from hyperemesis gravidarum and even had severe vomiting complicated by hypokalemia 3 d before delivery. The blood gas analysis of the twins at 2 h after birth showed severe metabolic alkalosis, hyponatremia, hypokalemia, hypochloremia and hyperlactatemia. Hyperglycemia appeared at 6 h after birth, and scleredema neonatorum at 24 h after birth. No significant abnormalities were found in the tandem mass spectrometer analysis of blood or urine samples. Whole-exome sequencing showed no abnormalities in the genes related to the phenotype. The twins were diagnosed with neonatal pseudo-Bartter syndrome. After symptomatic and supportive treatment, metabolic alkalosis and electrolyte disorders in the twins were completely resolved 4 d after birth. They were cured and discharged 51 d after birth without recurrence. Follow-up revealed no abnormalities in the physical or neurological development of the twins at 11 months after birth. Conclusions:Maternal hyperemesis gravidarum can lead to neonatal pseudo-Bartter syndrome, characterized by severe metabolic disorders as well as respiratory and circulatory dysfunction at the early stage after birth. Timely diagnosis and treatment are conducive to good prognosis in the affected neonates.

Recently, more research about the plant bioreactor expressing genes encoding human proteins was reported. In the present study, the cDNA of the human gene keratinocyte growth factor 2 (KGF2) was replaced with plant preferred codons by PCR, and the modified full-length cDNA was cloned into the plant expression vector pCAMBIA-YO containing the oil-body promoter. The fusion construct pCAMBIA-YO-KGF2 was transformed into Brassica napus by Agrobacterium tumefacien-mediated cotyledon transformation method. The transgenic seedlings were identified by PCR, Southern and western blot analysis all showed that KGF2 gene was successfully expressed in in transgenic Brassica napus.
Brassica napus ; genetics ; metabolism ; Cloning, Molecular ; DNA, Complementary ; genetics ; Fibroblast Growth Factor 7 ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Humans ; Plants, Genetically Modified ; genetics ; Rhizobium ; genetics ; Transformation, Genetic