To explore the mechanism of tea albino variation and high theanine formation, 'Fuyun 6' and a new theanine-rich tea cultivar 'Fuhuang 2' were as materials in this study, pigment content, metabolome and transcriptome of the two cultivars were analyzed by ultramicroelectron microscopy, widely targeted metabolomics, targeted metabolomics and transcriptomics. The results showed that five catechins, theobromine, caffeine, and 20 free amino acids, including theanine, glutamine, arginine, etc., were identified by targeted metabolomics. The amino acid content of 'Fuhuang 2' was significantly higher than that of 'Fuyun 6', and the theanine content was as high as 57.37 mg/g in 'Fuhuang 2'. The ultrastructure of leaves showed that the chloroplast cell structure of 'Fuhuang 2' was fuzzy, most of the grana lamellae were arranged in disorder, with large gaps, and the thylakoids were filiform. The determination of pigments showed that compared with 'Fuyun 6', the contents of chlorophyll A and B, carotenoids, flavonoids and other pigments of 'Fuhuang 2' decreased significantly, some important pigment-related-genes, such as chlorophyllase (CLH), 9-cis-epoxycarotenoid dioxygenase (NCED), flavonoid 3β-hydroxylase (F3H) and flavonoid 3', 5'-hydroxylase (F3'5'H) were significantly changed. Compared with 'Fuyun 6', 'Fuhuang 2' identified 138 significantly changed metabolites (SCMs) and 658 differentially expressed genes (DEGs). KEGG enrichment analysis showed that SCMs and DEGs were significantly enriched in amino acid biosynthesis, glutathione metabolism and TCA cycle. In general, the albino phenotype of 'Fuhuang 2' may be caused by a deficiency in photosynthetic proteins, chlorophyll metabolism genes and chlorophyll content. The accumulation of high theanine in 'Fuhuang 2' may be due to the low nitrogen consumption in yellowed leaves and the lack of carbon skeleton, amino and nitrogen resources are stored more effectively, resulting in the up regulation of metabolites and related gene expression in the amino acid synthesis pathway, theanine has become a significant accumulation of nitrogen-containing compounds in yellowed leaves.
Camellia sinensis/genetics* ; Chlorophyll A/metabolism* ; Plant Proteins/genetics* ; Plant Leaves/chemistry* ; Chlorophyll/metabolism* ; Transcriptome ; Flavonoids/metabolism* ; Amino Acids/genetics* ; Tea ; Mixed Function Oxygenases/metabolism* ; Nitrogen/metabolism*

Objective:To evaluate the safety and efficacy of Delorme procedure for adults with full-thickness rectal prolapse.Methods:Clinical data of 17 adult patients suffering from full-thickness rectal prolapse undergoing Delorme procedure from June 2014 to May 2018 in Hangzhou Third Hospital were retrospectively analyzed. Patient characteristics, operative data, postoperative complications, recurrence of rectal prolapse, continence state and constipation state were evaluated.Results:Eleven patients were female, 6 patients were male with a mean age of (68 ± 9) years. Operations were successfully performed in these 17 cases. The operation time was (88 ± 16) minutes. The estimated blood loss during operation was (23 ± 9) ml. The postoperative time of hospital stay was (8 ± 1) d. Two complications in two patients were observed. There was no treatment related death. One recurrent case was observed during (16 ± 2) months follow-up. The preoperative and postoperative mean constipation score of five patients with fecal constipation were (23 ± 2) and (11 ± 3) respectively ( t = 9.51, P<0.01). The mean fecal incontinence score of six patients with fecal incontinence, before and after Delorme procedure, were (14 ± 2) and (6 ± 2) respectively ( t = 9.09, P<0.01). Conclusions:The Delorme procedure for adults with full-thickness rectal prolapse is a safe and effective surgery with less complications and low recurrence rate. The Delorme procedure may be one of the preferred option of perineal approach for adults with full-thickness rectal prolapse, but the long-term outcome of Delormer procedure and its effect on postoperative anal function need to be further studied.

Objective:To investigate the interventional effect of metformin on pulmonary inflammation and pulmonary fibrosis in silicotic rats.Methods:In April 2019, 48 Wistar male rats of SPF grade were randomly divided into negative control group, metformin control group, silicon dioxide (SiO 2) model group, low, medium and high dose metformin intervention group according to the random number table method, 8 rats in each group. The SiO 2 model group and the low, medium and high dose metformin intervention groups were given 1 ml 50 mg/ml of SiO 2 by intratracheal instillation, the negative control group and the metformin control group were given 1 ml normal saline by intratracheal instillation. 24 hours later, the low, medium and high dose metformin intervention groups and the metformin control group were treated with 100, 200, 400 and 400 mg/kg metformin daily, the control and SiO 2 model groups received normal saline daily. Then the rats were sacrificed at the 28th day after SiO 2 exposure. The changes of rat body weight and pathological examination of rat lung tissue were observed, and the lung organ coefficient, the content of hydroxyproline (HYP) , the expression levels of inflammatory factors transforming growth factor beta1 (TGF-β1) , tumor necrosis factor-alpha (TNF-α) , interleukin-1beta (IL-1β) and the protein expression of E-cadherin (E-Cad) , Vimentin, α-SMA were detected. Results:Compared with the negative control group, SiO 2 model group had a significant decrease in the body weight of rats ( P<0.05) , lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-β1, TNF-α, IL-1β were all significantly increased ( P<0.05) . Compared with the SiO 2 model group, the weights of the rats in the medium and high dose intervention group of metformin increased significantly ( P<0.05) . And after intervention with different doses of metformin, the lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-β1, TNF-α and IL-1β were significantly decreased ( P<0.05) . Immunohistochemistry and Western blotting results showed that compared with the negative control group, the expression of E-Cad of the SiO 2 model group was decreased, and the expression levels of Vimentin and α-SMA were significantly increased ( P<0.05) . After metformin intervention, the expression of E-Cad was significantly increased, the expression levels of Vimentin and α-SMA were significantly decreased ( P<0.05) . Conclusion:Metformin can reduce lung tissue inflammation and fibrosis in rats exposed to SiO 2 dust, which may be related to reducing the expression of inflammatory factors in lung tissue and inhibiting the EMT process.

Objective:To explore the prevalence and influencing factors of allergic conjunctivitis in Baotou area at different times.Methods:A cross-sectional study was conducted in January 2021, 111 patients with allergic conjunctivitis who were admitted to the Department of Ophthalmology, the Second Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology in 2020 were selected as the research subjects. The basic information, allergic conditions, living habits and other information of the patients were obtained by self-made questionnaire. The pollen concentration and allergy related indicators(allergic duration and frequency, eye itching score, eye burn score, eye foreign body sensation score, tears score and the total scores) of 2019 and 2020 were analyzed by paired sample t-test. The pollen concentration of 2019 and 2020 were analyzed by Wilcoxon signed rank test. The allergy related indicators of different genders were analyzed by independent sample t-test. The allergy related indicators of different ages were analyzed by one-way analysis of variance. The relationship between the length of wearing masks and the allergy related indicators were analyzed by Spearman correlation analysis. Results:Among the 111 patients with allergic conjunctivitis, 54 were males and 57 were females, ranging in age from 8 to 69 years old, with a median age of 35 years old and an average age of (36.77±13.18) years old. The average pollen concentration in Baotou in 2019 was (125.35±222.64) grains/1 000 square millimeter, and the average pollen concentration in Baotou in 2020 was (107.38±137.29) grains/1 000 square millimeter. There was no significant difference in pollen concentration between the two years( Z=-0.178, P=0.859). The severity of allergic indicators in all patients in 2020 is significantly lower than in 2019( t values were 4.701,3.587,2.582,3.661,4.444,2.784,2.555,3.886,respectively, with P<0.05). The severity of allergic indicators in male patients is lower than that of female patients ( t values were -1.558, -1.257, -3.41, -3.085, -2.335, -2.897, -2.652, -4.124, respectively). The prevalence of allergic conjunctivitis dose not vary significant with age ( P values were 0.504, 0.095, 0.499, 0.265, 0.284, 0.655, 0.421, 0.976, respectively). In 2020, the average time that patients wear masks is (6.55±3.28) h/d, and there is a correlation with the difference in allergy duration ( r=0.191, P=0.045). Conclusion:The severity of allergic conjunctivitis in Baotou in 2020 was significantly improved compared with that in 2019, which may be related to masks wearing.

Objective:To explore the prevalence and influencing factors of allergic conjunctivitis in Baotou area at different times.Methods:A cross-sectional study was conducted in January 2021, 111 patients with allergic conjunctivitis who were admitted to the Department of Ophthalmology, the Second Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology in 2020 were selected as the research subjects. The basic information, allergic conditions, living habits and other information of the patients were obtained by self-made questionnaire. The pollen concentration and allergy related indicators(allergic duration and frequency, eye itching score, eye burn score, eye foreign body sensation score, tears score and the total scores) of 2019 and 2020 were analyzed by paired sample t-test. The pollen concentration of 2019 and 2020 were analyzed by Wilcoxon signed rank test. The allergy related indicators of different genders were analyzed by independent sample t-test. The allergy related indicators of different ages were analyzed by one-way analysis of variance. The relationship between the length of wearing masks and the allergy related indicators were analyzed by Spearman correlation analysis. Results:Among the 111 patients with allergic conjunctivitis, 54 were males and 57 were females, ranging in age from 8 to 69 years old, with a median age of 35 years old and an average age of (36.77±13.18) years old. The average pollen concentration in Baotou in 2019 was (125.35±222.64) grains/1 000 square millimeter, and the average pollen concentration in Baotou in 2020 was (107.38±137.29) grains/1 000 square millimeter. There was no significant difference in pollen concentration between the two years( Z=-0.178, P=0.859). The severity of allergic indicators in all patients in 2020 is significantly lower than in 2019( t values were 4.701,3.587,2.582,3.661,4.444,2.784,2.555,3.886,respectively, with P<0.05). The severity of allergic indicators in male patients is lower than that of female patients ( t values were -1.558, -1.257, -3.41, -3.085, -2.335, -2.897, -2.652, -4.124, respectively). The prevalence of allergic conjunctivitis dose not vary significant with age ( P values were 0.504, 0.095, 0.499, 0.265, 0.284, 0.655, 0.421, 0.976, respectively). In 2020, the average time that patients wear masks is (6.55±3.28) h/d, and there is a correlation with the difference in allergy duration ( r=0.191, P=0.045). Conclusion:The severity of allergic conjunctivitis in Baotou in 2020 was significantly improved compared with that in 2019, which may be related to masks wearing.

Objective:To investigate the interventional effect of metformin on pulmonary inflammation and pulmonary fibrosis in silicotic rats.Methods:In April 2019, 48 Wistar male rats of SPF grade were randomly divided into negative control group, metformin control group, silicon dioxide (SiO 2) model group, low, medium and high dose metformin intervention group according to the random number table method, 8 rats in each group. The SiO 2 model group and the low, medium and high dose metformin intervention groups were given 1 ml 50 mg/ml of SiO 2 by intratracheal instillation, the negative control group and the metformin control group were given 1 ml normal saline by intratracheal instillation. 24 hours later, the low, medium and high dose metformin intervention groups and the metformin control group were treated with 100, 200, 400 and 400 mg/kg metformin daily, the control and SiO 2 model groups received normal saline daily. Then the rats were sacrificed at the 28th day after SiO 2 exposure. The changes of rat body weight and pathological examination of rat lung tissue were observed, and the lung organ coefficient, the content of hydroxyproline (HYP) , the expression levels of inflammatory factors transforming growth factor beta1 (TGF-β1) , tumor necrosis factor-alpha (TNF-α) , interleukin-1beta (IL-1β) and the protein expression of E-cadherin (E-Cad) , Vimentin, α-SMA were detected. Results:Compared with the negative control group, SiO 2 model group had a significant decrease in the body weight of rats ( P<0.05) , lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-β1, TNF-α, IL-1β were all significantly increased ( P<0.05) . Compared with the SiO 2 model group, the weights of the rats in the medium and high dose intervention group of metformin increased significantly ( P<0.05) . And after intervention with different doses of metformin, the lung organ coefficient, alveolitis and fibrosis scores, HYP content and the levels of TGF-β1, TNF-α and IL-1β were significantly decreased ( P<0.05) . Immunohistochemistry and Western blotting results showed that compared with the negative control group, the expression of E-Cad of the SiO 2 model group was decreased, and the expression levels of Vimentin and α-SMA were significantly increased ( P<0.05) . After metformin intervention, the expression of E-Cad was significantly increased, the expression levels of Vimentin and α-SMA were significantly decreased ( P<0.05) . Conclusion:Metformin can reduce lung tissue inflammation and fibrosis in rats exposed to SiO 2 dust, which may be related to reducing the expression of inflammatory factors in lung tissue and inhibiting the EMT process.

OBJECTIVETo investigate the association between serum miRNA-6086 expression level and anal fistula recurrence.

METHODSClinical data and serum samples of 60 patients with anal fistula and mix hemorrhoid identified by pathology undergoing resection in our department from August 2015 to August 2016 were collected. In addition, serum samples of 20 patients matching with fistula group in age, gender and body weight and receiving only hemorroidectomy were collected as control during the same period. Serum miRNA6086 expression level was detected by real-time quantitative RT-PCR method, and the association of serum miRNA6086 expression level with clinicopathologic features was analyzed. Univariate ANOVA test and multivariate logistic regression analysis were performed to analyze the association between serum miRNA6086 expression level and anal fistula recurrence.

RESULTSThe relative expression of serum miRNA6086 in fistula group was 65.85±15.57, which was significantly up-regulated for 4.87 folds of 13.52±7.32 in control group(P<0.05). In fistula group, 24 cases(40%) developed anal fistula recurrence, whose serum miRNA6086 expression was significantly higher compared to 36 cases without recurrence (74.06±12.92 vs. 60.38±14.90, P<0.05). No associations of serum miRNA6086 expression with age, gender, BMI, drug history, acute phase were observed (P>0.05), while association of serum miRNA6086 expression level with the type, number and position of anal fistula was significant (all P<0.05). Univariated analysis indicated that anal fistula type (χ=6.890, P=0.009), anal fistula number (χ=0.554, P<0.001) and serum miRNA6086 expression (χ=11.390, P=0.001) were significantly associated with anal fistula recurrence. Multivariate logistic regression analysis showed that complex anal fistula (OR=4.75, 95%CI: 1.84 to 12.01, P=0.001) and high expression of serum miRNA6086 (OR=3.22, 95%CI:1.31 to 8.22, P=0.011) were independent risk factors of anal fistula recurrence.

CONCLUSIONUp-regulated expression of serum miRNA6086 is associated to the anal fistula type and may be valuable in predicting the prognosis.

OBJECTIVETo investigate the expression of prostaglandin transporter (PGT) in colorectal cancer (CRC) tissues and its relationship with clinicopathological features.

METHODSThe mRNA and protein levels of PGT were determined by real-time PCR, Western blot and immunohistochemical methods in cancer tissues and adjacent normal tissue from 80 patients with colorectal cancer and their relationship with clinicopathological features was analyzed.

RESULTSCompared with the adjacent normal tissue of colorectal cancer, the PGT mRNA relative expression (0.57 ± 0.33 vs. 2.33 ± 1.20) and the PGT protein expression in cancer tissues decreased significantly [PGT/GAPDH 0.45 ± 0.16 vs. 0.78 ± 0.23, integral A 718.7 ± 359.4 vs. 10412.0 ± 6423.3, average A 0.03 ± 0.01 vs. 0.12 ± 0.09, all P<0.01]. Lower mRNA and protein expressions of PGT in colorectal cancer were associated with depth of invasion T3 to T4 and TNM stage III( to IIII( (P<0.01), while not associated with gender, age, tumor location and differentiation degree (all P>0.05).

CONCLUSIONExpression levels of PGT mRNA and protein in colorectal cancer tissue are significantly down-regulation. PGT expression is associated with invasion depth and late stages.

Colorectal Neoplasms ; Down-Regulation ; Humans ; Neoplasm Invasiveness ; Neoplasm Staging ; Organic Anion Transporters ; RNA, Messenger

OBJECTIVETo investigate the effects of intragastric administration of human interferon-α (hIFN-α)-transformed Bifidobacterium on immune functions of mice.

METHODSThe E.coli-Bifidobacterium shuttle expression vector containing hIFN-α gene was constructed and transformed into Bifidobacterium. The hIFN-α-transformed Bifidobacterium suspension (1010 /ml) was prepared after induction with 0.2% L-arabinose for hIFN-α expression and administered intragastrically in male Balb/C mice at the dose of 0.1 ml every other day for 2 weeks, with the mice receiving empty vector-transformed Bifidobacteria as the negative control and those having an equal volume of saline as the blank control. The percentages of mononuclear cell subsets in the thymus, spleen and blood were detected in the mice by flow cytometry, and the serum levels of IL-4, IL-12, IFN-γ and TNF-α were assayed using mouse cytokine FlowCytomix Kit.

RESULTSThe percentages of CD3⁺CD8⁺ and CD4⁺CD8⁺ cells in the thymus, CD3⁺CD4⁺, CD3⁺CD8⁺ and CD4⁺CD8⁺ cells in the spleen, and CD3⁺CD8⁺ cells in the blood all increased significantly in IFN group as compared with those in the negative and blank control groups (P<0.01 or 0.05). The serum level of IFN-γ also increased significantly (P<0.05) while IL-4 level remained unchanged in IFN group compared with those in the two groups.

CONCLUSIONIntragastric administration of hIFN-α-transformed Bifidobacterium promotes lymphocyte proliferation and maturation and increases the serum levels of Th1 cytokines in mice.

Animals ; Bifidobacterium ; Cell Proliferation ; Genetic Vectors ; Humans ; Interferon-alpha ; pharmacology ; Interferon-gamma ; blood ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; pharmacology ; Spleen ; cytology ; Th1 Cells ; cytology ; Thymus Gland ; cytology ; Tumor Necrosis Factor-alpha ; blood

Objective To investigate the effects of intragastric administration of human interferon-α (hIFN-α)-transformed Bifidobacterium on immune functions of mice. Methods The E.coli-Bifidobacterium shuttle expression vector containing hIFN-αgene was constructed and transformed into Bifidobacterium. The hIFN-α-transformed Bifidobacterium suspension (1010/ml) was prepared after induction with 0.2%L-arabinose for hIFN-αexpression and administered intragastrically in male Balb/C mice at the dose of 0.1 ml every other day for 2 weeks, with the mice receiving empty vector-transformed Bifidobacteria as the negative control and those having an equal volume of saline as the blank control. The percentages of mononuclear cell subsets in the thymus, spleen and blood were detected in the mice by flow cytometry, and the serum levels of IL-4, IL-12, IFN-γand TNF-αwere assayed using mouse cytokine FlowCytomix Kit. Results The percentages of CD3+CD8+and CD4+CD8+cells in the thymus, CD3+CD4+, CD3+CD8+and CD4+CD8+cells in the spleen, and CD3+CD8+cells in the blood all increased significantly in IFN group as compared with those in the negative and blank control groups (P<0.01 or 0.05). The serum level of IFN-γalso increased significantly (P<0.05) while IL-4 level remained unchanged in IFN group compared with those in the two groups. Conclusion Intragastric administration of hIFN- α- transformed Bifidobacterium promotes lymphocyte proliferation and maturation and increases the serum levels of Th1 cytokines in mice.