This study was aimed to observe the intervention effect of oxidation/antioxidation at different time point among mice induced by lipopolysaccharide (LPS) and bleomycin. It provided experimental basis for the establishment of chronic obstructive pulmonary disease (COPD) animal model with qi-deficiency and phlegm-obstructing pattern with inducers mentioned above. A total of 96 mice were randomly divided into the normal control group, bleomycin group, and LPS group, with 32 mice in each group. In the bleomycin group and LPS group, 40 μL of nasal drops were given with bleomycin at the concentration of 3.75 μg/μL or LPS at the concentration of 5 μg/μL, respectively to establish the COPD animal model with qi-deficiency and phlegm-obstructing pattern. On the 1st day, 7th day, 14th day and 28th day after the model establishment, the general status and activities of mice were recorded. And traditional Chinese medicine (TCM) signs such as skin color of the four limbs, skin color under the tongue and color of the tail were also collected when the animal model was sacrificed. At each time point, 8 mice were sacrificed. The lung tissues were removed. And the contents of GSH, MDA, SOD and T-AOC were detected in the homogenate of lung tissues. The results showed that compared with the normal control group, mice in the bleomycin group had slightly dull eyes, dry hair without burnish, upright and fluffy hair, dark purple skin color of the auricle and four claws, tiredness, inactivity, occasional cough, asthma or rapid breathing. The GSH content of lung tissues on the 7th day, 14th day and 28th day was obviously reduced (P< 0.05, or P< 0.01). The MDA, SOD and T-AOC contents on the 1st day, 7th day, 14th day and 28th day were obviously reduced (P< 0.05, or P< 0.01). Compared with the normal control group, mice in the LPS group had slightly dull eyes, soft hair with slight burnish, pale red skin color of the auricle and four claws, tiredness; some mice preferred to gather. Contents of GSH and SOD in lung tissues on the 1st day and 7th day were obviously reduced (P< 0.05, or P< 0.01). Contents of MDA and T-AOC on the 1st day, 7th day and 14th day were obviously reduced (P < 0.05, or P < 0.01). It was concluded that obvious oxidation/antioxidation imbalance started on the 7th day in lung tissues of mice in the bleomycin group. It reduced later on. And the oxidation/antioxidation imbalance continued until the end of the model establishment. Obvious oxidation/antioxidation imbalance started on the 1st day in lung tissues of mice in the LPS group. However, this oxidation/antioxidation imbalance was adjusted back to normal level through time.

Objective: To analyze the mutations of BRCA1 in 9 Chinese familiar breast cancer patients. Methods: Peripheral blood samples were obtained from 9 patients enrolled from 9 breast cancer families, one normal control, 32 sporadic breast cancer patients and 33 normal donors. DNA extracted from lymphocytes was amplified by polymerase chain reaction (PCR). The 22 exons and partial introns of BRCA1 were screened by PCR denaturing high performance liquid chromatography (PCR-DHPLC) and confirmed by direct sequencing. Results: Among these 9 familiar breast cancer patients, a deleterious mutation was detected in one case in exon 11 (3870delTGTC) which was a 4 base deletion and caused a frameshift in turn. One novel and unique amino acid substitution (E867R) was detected in one case. Eight patients were detected to have a known variation in intron 18 (IVS18+65G→A), and the ratio of this variation detected was 88.9%(8/9). The ratio of this variation was 37.5%(12/32) in sporadic breast cancer patients or 33.3%(11/33) in normal control. This variation was found to be accompanied all the time with a known missense variation in exon 11 (P871L) and a polymorphism in intron 9 (IVS8 57delT). Those three variants were also detected in homozygous in one case, which implies the linkage of the 3 sites. The linkage had not been reported. Two patients had been found with a known polymorphism in exon 13 (S1436S). Another known polymorphism was found in one case (L771L). In addition, intronic variants (IVS2+48C→T, IVS2+133C→T, IVS12+112C→A) were detected. Conclusion: The mutations of BRCA1 in Chinese familiar breast cancer patients are different from the hot spots reported in Caucasian and Jewish. It is important that further study be conducted to seek for specific mutations of this gene or other possible relevant genes in Chinese familiar breast cancer patients.

Objective To study the pathogeny,clinical manifestation,diagnosis and treatment of emergency cases admitted to endocrine department with hypoglycemia.Methods The clinical data of 37 emergency cases with hypoglycemia admitted to endocrine department were retrospectively analyzed.Results 37 patients with emergency hypoglycemia were admitted to the Central Hospital of Luohe.34 cases were classified as diabetes,3 cases without diabetes.Among 37 patients,35 cases were recovered and discharged after treatment,1 case became vegetative,1 case died at last.Conclusion To strengthen blood glucose monitoring on the emergency patients with hypoglycemia is nec-essary.We must diagnose and treat hypoglycemia in time.Meanwhile,we should offer health education for diabetic patients,and the hypoglycemic agent or insulin must be reasonably applied to avoid hypoglycemia.

Objective To develop a rapid and sensitive DNA microarray for Listeria monocytogenes detection.Methods A DNA microarray was developed using gyrB,ISR,16S rRNA,23S rRNA,hlyA,iap and prfA as the target genes and tested against 18 different species of known reference for repeatability,sensitivity,and specificity to verify the effectiveness of the chip.Results After testing of samples by the LM array,results show that the 70 mer Oligos synthesized by IDT are superior to the Oligos synthesized by Sagon with respect to both probe spotting or samples detection.The comparison of 3 spotting probe concentrations of 10 μmol/L,40 μmol/L and 80 μmol/L demonstrated that the 10 pmol/L probes result in good detection signals equivalent to the 40 μmol/L and 80 μmol/L probes.The repeatability and sensitivity evaluated by sample testing on the LM array revealed that the chips developed in this study have good repeatability and the lower limit of sample detection is 0.9 ng DNA.The LM array can distinguish clearly and definitively between Listeria and non-Listeria bacteria in the sample.Conclusion The microarray is able to rapidly detect and identify Listeria monocytogenes.

This study was aimed to reveal the effects and molecular regulation mechanism of methyl jasmonate (Me-JA) on volatile terpenoids from Amomum villosum Lour. After the leaves and fruits of A momum villosum Lour. were treated with different concentrations of MeJA, the volatile terpenoids of fresh fruits from A . villosum Lour. were ex-tracted with microwave method and analyzed by GC-MS. Then, leaves and fruits treated with MeJA were sequenced by Illumina. The transcriptome data was analyzed by bioinformatic methods. The results showed that there were 20 and 33 volatile terpenoids detected in peels and seed groups, respectively. Contents of volatile terpenoids in peels and seed groups were both improved after 600 μmol·L-1 MeJA treating fruits for 24 h, such as bornyl acetate, cam-phor, borneol, and etc. While 200 μmol·L-1 MeJA treating different parts for 24 h can regulate the biosynthesis of some volatile terpenoids in peels differently. And 200 μmol·L-1 MeJA treating fruits can improve the content of ma-jor volatile terpenoids in seed groups. A total of 68 168 unigenes were obtained with de novo assembly, and 48 627 unigenes were annotated after comparison with public protein databases. Analysis of functional annotation against KEGG database showed that there were 208 unigenes closely related with metabolism of volatile terpenoids and 22 u-nigenes related with MYC2 transcription factors. It was concluded that MeJA can effectively regulate the metabolism of volatile terpenoids from A . villosum Lour. There were a lot of candidate genes related with the biosynthesis of volatile terpenoids obtained by analyzing the transcriptome data which also provided a large amount of data for the discovery and regulation of functional genes related with the biosynthesis of volatile terpenoids from A . villosum Lour.

Objective To understand the genetic characteristics of Enterovirus 71 ( EVT1 ) circu-lating strains of Guangdong province in 2008. Methods We isolated an EV71 strain from the fatal case of the hand, foot and mouth disease during an epidemic of Guangdong in 2008. Its complete genome was se-quenced and analyzed comparatively. Results The results showed that the full length of EV71 GDFS-3 ge-nome( not including poly A tail ) is 7405 bp. No insertion or deletion is detected in the coding region. There are several insertions and deletions in 5'and 3'UTR. Phylogenetic analysis of GDFS-3 and reference strains showed GDFS-3 strain shares the highest nueleotide homology with TW984 strain(96.0% ) but low homology with SIN5865, MS and BrCr( about 81.0% ). GDFS-3 strain also shares the highest amino acid homology with TW984 strain(99.0% ). It clustered with reference strains of CA subgenotype in the phylogenetie tree. The nucleotide identity with CA reference strains is 91.0% -95.0%. Conclusion The phylogenetic analysis based on the entire genome demonstrates that GDFS-3 strain has the nearest genetic relationship with TW984 strains ( isolated in 2004). GDFS-3 may belong to the same subgenogroup ( CA ) with Taiwan predominant strains. Otherwise,Some mutations in 5'UTR of EV71 may play the very important role in heightened viru-lence.

The aim of this study was to investigate the effects of Avastin on aquaporin4 (AQP4) expression in human retinal Müller cells in vitro under hypoxia, so as to explore the mechanism of Avastin treating retinal edema. The human Müller cells were cultured using the enzymatic digestion method. Müller cells were identified under the transmission electron microscopy and by using immunofluorescence staining. By using semi-quantitative reverse transcription polymerase chain reaction (RT-PCR), the expression of AQP4 mRNA and VEGF mRNA in Müller cells cultured with 500 μmol/L CoCl(2) for 0, 3, 6, 12 and 24 h, and with 0, 100, 300, 500 and 700 μmol/L CoCl(2) for 24 h was detected. The expression of AQP4 mRNA in Müller cells cultured with 50 ng/mL exogenous vascular endothelial growth factor (VEGF) for 0, 0.5, 1, 2 and 4 h, and with 0, 25, 50 and 75 ng/mL VEGF for 24 h was detected. Amplified cDNA products of AQP4 mRNA in Müller cells cultured with 500 μmol/L CoCl(2) and 200 μg/mL Avastin for 24 h were detected. The results showed that more than 95% cells displayed positive immunofluorescence reaction. Characteristic 8-10 nm intracellular filaments could be seen in the cytoplasm under the transmission electron microscopy. In the CoCl(2) experimental groups, the expression of AQP4 mRNA and VEGF mRNA in Müller cells was increased as compared with the control group. Alteration of AQP4 mRNA and VEGF mRNA levels showed a significantly positive correlation (r (2)=0.822, P<0.05). The expression of AQP4 mRNA in Müller cells was increased by VEGF. The expression of AQP4 mRNA was significantly decreased by Avastin as compared with the control group. It is suggested that Avastin can decrease the expression of AQP4 mRNA in human Müller cells under chemical hypoxic conditions partially via VEGF path, which may be one of the mechanisms of Avastin treating retinal edema.

Objective To identify the reliable reference genes for gene expression analysis of the pericarp and seed of Amomum villosum Lour. by using real-time fluorescence quantitative polymerase chain reaction ( qRT-PCR). Methods Using the fruits ( separated into peels and seeds) of A. villosum at three different developmental periods as the experimental material, 5 candidate reference genes (β-actin, EF-1α, GAPDH, PGK, TUA) with steady expression were screened out by the high throughout sequencing of transcriptome and expression profile data. The qRT-PCR technique was applied to study the expression levels of 5 candidate reference genes in different samples. The stability of the candidate reference genes were evaluated by GeNorm and NormFinder software. Results The 5 reference genes had different stabilities in the pericarp and seed of A. villosum Lour. at different development periods . The order of the steadiness of reference genes showed by GeNorm was EF-1α = TUA>PGK>GAPDH>β-actin. The results of NormFinder revealed that EF-1α was the most stable, followed by TUA, and the order of the other three genes was as same as the results of GeNorm. Conclusion EF-1αand TUA could be used as double reference genes for the normalization of gene expression in A. villosum fruits at different developmental periods by using qRT-PCR.

Objective To investigate the prevalence and risk factors of mild cognitive impairment (MCI) among the old people in Chongqing, China. Methods From October to November, 2015, 1850 persons more than 60-year-old in Chongqing were cluster sampled. They were investigated with self-made general situation questionnaire, Mini-Mental State Examination (MMSE), Instrumental Activity of Daily Living Scale (IADL) and Geriatric Depression Scale (GDS) through face-to-face interview. Enumeration data were tested withχ2 test and multiple Logistic regression. Results MCI prevalence was 11.73%in the old people in Chongqing, and various with the residential re-gions, ages, marital status, literacy, career, smoking, drinking, seafood-eating, egg-eating, exercising, intensive labor, low intensity of labor, housework, outdoor activities, Mahjong/chess/cards playing, TV-watching/radio-listening/newspaper-reading, social activities, obesity, hy-pertension, diabetes, stroke, hyperglycemia, cerebrovascular insufficiency and depression (χ2>4.092, P<0.05). According to multiple Logis-tic regression, living in rural areas, aging, celibacy, obesity, hypertension and depression were the risk factors of MCI;while middle school and above years of schooling, mental workers, smoking cessation, frequent seafood-eating and egg-eating, exercising, daily intensive labor and low intensity of labor, daily and sometimes housework, daily and weekly outdoor activities at least once, daily Mahjong/chess/cards playing and daily TV-watching/radio-listening/newspaper-reading were the preventing factors. Conclusion The morbidity of MCI is high in the old people in Chongqing. It is necessary to take effective intervention in view of the related factors of MCI as early as possible, to delay or prevent the development of MCI to dementia.

Objective: To understand the relationship between lipid metabolism and heart rate deflection point (HRDP) in Tibetan children in the high altitude area of Ganzi, and to provide reference for effective obesity prevention and physical exercise intervention for Tibetan children. Methods: From September to October 2019, 284 Tibetan primary school students living in Ganzi area were randomly selected. HRDP and deflection point speed with different body mass index (BMI) by increasing load method, as well as serum lipid metabolism were assessed. The correlation between the HRDP and lipid metabolism level was analyzed. Results: There were statistically significant differences in serum leptin, adiponectin, total cholesterol, and triglyceride levels among the first and second grade Tibetan primary school students in the Ganzi high altitude area with different nutritional status ( F =22.16, 10.12, 11.24, 4.35, P <0.05). The heart rate values at the HRDP in the first and second grades, the third and fourth grades, and the fifth and sixth grades under different nutritional status were compared, and the differences were statistically significant ( F =3.35, 4.76, 4.68, P <0.05). Pearson correlation analysis showed that there were statistically significant correlations between HRDP and serum leptin in the obese, overweight, normal, and lean groups in the first and second grades of primary school ( r =0.66, 0.14, 0.45, 0.65 , P <0.05). Conclusion There is a close relationship between HRDP and lipid metabolism of Tibetan children in the high altitude area of Ganzi, and the heart rate at the deflection point can be used as an effective reference intensity for exercise intervention in plateau children.