Current status and future of treatment of pulmonary fibrosis by mesenchymal stem cells and extracellular vesicles / 中国组织工程研究

BACKGROUND:Despite a series of clinical treatment measures,the treatment of pulmonary fibrosis still faces challenges.In recent years,mesenchymal stem cells and their extracellular vesicles have attracted extensive attention as an emerging therapeutic strategy and are considered to be a promising means of treating pulmonary fibrosis. OBJECTIVE:To systematically review the application of mesenchymal stem cells and their extracellular vesicles in the treatment of pulmonary fibrosis,to comprehensively understand their therapeutic mechanism,efficacy evaluation and problems,and provide reference and guidance for further research and clinical application in the future. METHODS:Using Chinese and English search terms"mesenchymal stem cells","mesenchymal stem cell extracellular vesicles","pulmonary fibrosis",we searched the CNKI and PubMed electronic journal databases.By means of manual reading and eliminating duplicate articles,112 articles were selected,but 58 Chinese and English articles were finally included for summary. RESULTS AND CONCLUSION:(1)Mesenchymal stem cells and their extracellular vesicles have shown great potential in the treatment of pulmonary fibrosis,such as regulating inflammatory responses,inhibiting fibroblast proliferation,and promoting damaged tissue repair.Preliminary results from clinical trials have also shown some effects of the treatment,including improved lung function and quality of life in patients.(2)However,mesenchymal stem cells and extracellular vesicles in the treatment of pulmonary fibrosis still face some challenges.During treatment,technical challenges such as cell migration and intrachistological localization need to be addressed for it to accurately reach the damaged lung tissue.Furthermore,its long-term safety also needs to be further studied and improved.For translational medicine development,standardized procedures such as cell collection,cell isolation,cell culture,cell harvesting,and cell identification need to be refined.(3)Despite these challenges,through the joint efforts of scientific researchers and medical personnel,these problems are expected to be gradually solved.In the future,we can further improve treatment outcomes by optimizing treatment regimens and exploring individualized treatments.At the same time,in-depth research on the therapeutic mechanism of stem cells and their extracellular vesicles is expected to develop more efficient and safe therapeutic strategies.

A machine learning-based predictive model of nonsuicidal self-injurious behavior among college students in Guizhou Province / 中国学校卫生

Objective@#To explore the effectiveness of machine learning algorithms in predicting non-suicidal self-injury (NSSI) behavior among college students, and to analyze the influencing factors of NSSI behavior, thus providing a reference for promoting psychological well-being.@*Methods@#In December 2022, a stratified random cluster sampling method was used to select 835 college students from a university in Guizhou Province, China. The Adolescent Self-injury Scale, Family Function Assessment Scale, and Emotion Regulation Self-efficacy Scale were used to evaluate the participants. Demographic characteristics, family factors, and emotional factors were taken as independent variables, while the dependent variable was whether college students exhibited NSSI behavior. Machine learning algorithms, including Logistic regression, support vector machine (SVM), decision trees, algorithm gradient boosting trees, random forests, and AdaBoost, were used to construct predictive models.@*Results@#The detection rate of NSSI behavior among the college students was 23.23% (194 individuals). The NSSI behavior group scored higher than the non-NSSI behavior group in total family function, emotional communication, egoism, and family rules ( t=3.02, 3.35 , 2.23,2.87, P <0.05). On the other hand, the non-NSSI behavior group scored higher than the NSSI behavior group in total emotion regulation selfefficacy, managing negative emotion self-efficacy, and expressing positive emotion self-efficacy ( t=-5.04, -5.48 , -2.43, P <0.05). The recall rates of random forests, SVM, Logistic regression, decision trees, algorithm gradient boosting trees, and AdaBoost were 84.3% , 90.6%, 73.4%, 87.5%, 95.3%, 89.0%, respectively. The F1 scores were 84.4%, 92.1%, 71.2 %, 79.4%, 91.7%, 89.1% , respectively. The respective precision rates were 84.4%, 93.5%, 69.1%, 72.7%, 88.4%, 89.1 %. The AUC scores were 0.845, 0.922, 0.706, 0.776, 0.915, and 0.891, respectively.@*Conclusion@#Compared to the algorithm gradient boosting tree, random forest, Logistic regression, and AdaBoost models, the SVM model has a better predictive effect on whether college students in Guizhou Province exhibits NSSI behavior. It is recommended to use an appropriate model to identify students at risk of NSSI behavior as early as possible and provide psychological crisis interventions to promote their mental health.

Chiral separation methods and the applications in the study of chiral components of traditional Chinese medicines / 药学学报

Chirality is one of the fundamental properties of nature, and most of the important molecules in living organisms contain chiral structures. The efficacy and safety of drugs are often closely related to the chiral structure of compounds, however, there are relatively more studies on synthetic characterization, pharmacology, and toxicology of chiral small molecule chemical drugs, but relatively less studies on chiral compounds contained in natural drugs such as traditional Chinese medicines. Chiral separation, as the basis of chiral research, has a pivotal position in the study of chiral compounds. In this paper, we systematically describe the separation methods of chiral compounds from the classification of chiral splitting methods based on chromatographic and non-chromatographic methods, as well as chromatographic packing materials, chiral additives and chiral derivatization, and review the chiral compounds in natural drugs such as traditional Chinese medicines reported in the past ten years, in order to provide references for the splitting and evaluating the activity of chiral compounds, and the improvement of quality standards of traditional Chinese medicines.

Analysis of surgical types for patients with Kashin-Beck disease in Shaanxi Province from 2018 to 2020 / 中华地方病学杂志

Objective:To analyze the surgical types of patients with Kashin-Beck disease (KBD) in Shaanxi Province, and to provide reference for optimizing KBD surgery.Methods:Retrospective analysis was used to investigate KBD patients who underwent surgical treatment, and the patient's information was from the Shaanxi Provincial Endemic Disease Prevention and Control Information System. Patients with incomplete surgical treatment information were excluded, and χ 2 test was used to analyze the impact of factors such as gender, education level, living area, disease severity, and social environment on the selection of different surgical types (free body enucleation or joint replacement) for KBD patients. Using propensity score method to perform a 1 ∶ 1 match on patients with different surgical types, and analyzing the influencing factors of KBD patients receiving different types of surgical treatment through multivariate logistic regression. Results:From 2018 to 2020, a total of 1 084 KBD patients in Shaanxi Province underwent surgical treatment, including 555 males (51.20%) and 529 females (48.80%), with a median age of 64 years old. There were 917 patients (84.59%) in the Guanzhong region (Tongchuan, Xi'an, Weinan, Xianyang, Baoji), 120 patients (11.07%) in the northern Shaanxi region (Yulin, Yan'an), and 47 patients (4.34%) in the southern Shaanxi region (Ankang, Hanzhong, Shangluo). The number of patients with KBD grade Ⅰ,Ⅱ, and Ⅲ was 401 (36.99%), 525 (48.43%), and 158 (14.58%), respectively. Five hundred and forty-eight patients (50.55%) underwent free body enucleation surgery, and 536 patients (49.45%) underwent joint replacement surgery. Univariate analysis showed that different genders, educational levels, living areas, disease severity, economic status, whether to relocate, type of drinking water, type of staple food, and source of staple food were the influencing factors for KBD patients to choose the type of surgery (χ 2 = 81.82, 22.38, 93.68, 22.38, 5.17, 15.68, 13.82, 39.37, 49.63, P < 0.05). Among 374 pairs of patients who underwent free body enucleation and joint replacement through propensity score matching, multivariate logistic regression analysis found that high school and above education level [odds ratio( OR) = 0.42, P = 0.008], Guanzhong region ( OR = 0.12, P < 0.001), relocation ( OR = 0.60, P = 0.004), and drinking well water ( OR = 2.15, P = 0.001) were the influencing factors for KBD patients to choose different surgical types for treatment. Conclusion:When performing surgical treatment for KBD patients in Shaanxi Province, clinicians can consider the severity of the disease, as well as factors such as the education level, living area, relocation status, and type of drinking water of KBD patients, to provide more suitable surgical types for KBD patients.

Identification and analysis of terpene synthase (TPS) gene family in Schizonepeta tenuifolia / 中国中药杂志

Terpenoids are important secondary metabolites of plants that possess both pharmacological activity and economic value. Terpene synthases(TPSs) are key enzymes in the synthesis process of terpenoids. In order to investigate the TPS gene family members and their potential functions in Schizonepeta tenuifolia, this study conducted a systematic analysis of the TPS gene family of S. tenuifolia based on the whole genome data of S. tenuifolia using bioinformatics methods. The results revealed 57 StTPS members identified from the genome database of S. tenuifolia. The StTPS family members encoded 285-819 amino acids, with protein molecular weights ranging from 32.75 to 94.11 kDa, all of which were hydrophilic proteins. The StTPS family members were mainly distributed in the cytoplasm and chloroplasts, exhibiting a random and uneven physical localization pattern. Phylogenetic analysis showed that the StTPS genes family were divided into six subgroups, mainly belonging to the TPS-a and TPS-b subfamilies. Promoter analysis predicted that the TPS gene family members could respond to various stressors such as light, abscisic acid, and methyl jasmonate(MeJA). Transcriptome data analysis revealed that most of the TPS genes were expressed in the roots of S. tenuifolia, and qRT-PCR analysis was conducted on genes with high expression in leaves and low expression in roots. Through the analysis of the TPS gene family of S. tenuifolia, this study identified StTPS5, StTPS18, StTPS32, and StTPS45 as potential genes involved in sesquiterpene synthesis of S. tenuifolia. StTPS45 was cloned for the construction of an prokaryotic expression vector, providing a reference for further investigation of the function and role of the TPS gene family in sesquiterpene synthesis.

A pilot study of plasma interleukin-6 and interleukin-27 in differential diagnosis of acute respiratory distress syndrome and neonatal respiratory distress syndrome in preterm infants / 中国当代儿科杂志

OBJECTIVES@#To study the significance of interleukin-6 (IL-6) and interleukin-27 (IL-27) in the differential diagnosis of acute respiratory distress syndrome (ARDS) and neonatal respiratory distress syndrome (NRDS) in preterm infants.@*METHODS@#The preterm infants with the manifestation of respiratory distress who were treated in the Neonatal Diagnosis and Treatment Center, Children's Hospital of Chongqing Medical University, from March to November 2021, were enrolled in this prospective study. According to the diagnosis results, they were divided into two groups: ARDS group (n=18) and NRDS group (n=20). ELISA was used to measure the plasma levels of IL-6 and IL-27. The receiver operating characteristic (ROC) curve was used to analyze the value of each index in the diagnosis of ARDS.@*RESULTS@#The ARDS group had significantly higher plasma levels of IL-6 and IL-27 than the NRDS group (P<0.05). The ROC curve analysis showed that IL-6 had an area under the ROC curve (AUC) of 0.867 for the diagnosis of ARDS, with a sensitivity of 61.1% and a specificity of 95.0% at the cut-off value of 56.21 pg/mL. The ROC curve analysis also showed that IL-27 had an AUC of 0.881 for the diagnosis of ARDS, with a sensitivity of 83.3% and a specificity of 80.0% at the cut-off value of 135.8 pg/mL.@*CONCLUSIONS@#Plasma IL-6 and IL-27 can be used as biological indicators for early differential diagnosis of ARDS and NRDS in preterm infants.

Metabolic dysregulation and emerging therapeutical targets for hepatocellular carcinoma

Hepatocellular carcinoma (HCC) is an aggressive human cancer with increasing incidence worldwide. Multiple efforts have been made to explore pharmaceutical therapies to treat HCC, such as targeted tyrosine kinase inhibitors, immune based therapies and combination of chemotherapy. However, limitations exist in current strategies including chemoresistance for instance. Tumor initiation and progression is driven by reprogramming of metabolism, in particular during HCC development. Recently, metabolic associated fatty liver disease (MAFLD), a reappraisal of new nomenclature for non-alcoholic fatty liver disease (NAFLD), indicates growing appreciation of metabolism in the pathogenesis of liver disease, including HCC, thereby suggesting new strategies by targeting abnormal metabolism for HCC treatment. In this review, we introduce directions by highlighting the metabolic targets in glucose, fatty acid, amino acid and glutamine metabolism, which are suitable for HCC pharmaceutical intervention. We also summarize and discuss current pharmaceutical agents and studies targeting deregulated metabolism during HCC treatment. Furthermore, opportunities and challenges in the discovery and development of HCC therapy targeting metabolism are discussed.

Analysis of monitoring results of Keshan disease in Shaanxi Province in 2020 / 中华地方病学杂志

Objective:To grasp the current situation of Keshan disease in Shaanxi Province, and to provide scientific basis for evaluation of local Keshan disease control and elimination.Methods:From May to November 2020, Keshan disease surveillance was carried out in all the diseased townships (towns) of 29 diseased counties (districts, cities) in Shaanxi Province. The diseased village was used as the unit to collect the demographic data of the diseased areas, and the village doctor's clues were used to investigate suspected myocardial cases, and recent outpatient or inpatient clinical examination data of Keshan disease cases were collected; suspected cases were organized for clinical physical examination, electrocardiography (ECG), chest X-ray and echocardiography examination. According to "Notice of the National Health Commission on Printing and Distributing the Evaluation Measures for the Control and Elimination of Key Endemic Diseases (2019 Edition)", the diseased compliance status was assessed.Results:There were 174 diseased townships (towns) and 2 653 diseased villages in the whole province, with a permanent population of 2 819 342 people. A total of 87 suspected cases and 29 confirmed cases of Keshan disease were found, including 27 cases of chronic Keshan disease and 2 cases of latent Keshan disease. The acute and subacute Keshan disease and new cases with disease duration less than 1 year were not detected. The ST-T changes, premature ventricular contractions, complete right bundle branch block, left ventricular hypertrophy, and atrial fibrillation were the most common ECG changes in 29 cases of Keshan disease, and the single ECG change accounted for 6.90% (2/29), two changes accounted for 44.83% (13/29), and three or more changes accounted for 48.28% (14/29). There were 1 case with normal cardiothoracic ratio in chest X-ray examination, 10, 13, and 4 cases with mild, moderate and severe enlargement, respectively; 13 cases with enlarged left atrium in echocardiography, 15 cases with enlarged left ventricle, and 6 cases with enlarged heart.Conclusions:All the diseased areas in Shaanxi Province have reached the national standard for elimination of Keshan disease. Keshan disease monitoring should be continued, self-management of patients with chronic Keshan disease and follow-up observation of patients with latent Keshan disease should be strengthened, and treatment programs for patients with Keshan disease should be actively carried out to improve the quality of life of the patients.

Effect of selenium supplementation on 10-year survival rate of chronic Keshan disease / 中华地方病学杂志

Objective:To observe the effect of selenium supplementation on 10-year survival rate of chronic Keshan disease (KD).Methods:The 10-year follow-up data of 302 patients with chronic KD at the KD surveillance sites in Shaanxi Province were collected from the Shaanxi Institute for Endemic Disease Control and Research and Xi'an Jiaotong University, 170 (56.3%) cases were given selenium supplementation (oral administration of sodium selenite tablet, once a week, 1 mg/time) until the end point of follow-up as selenium supplementation group, and the rest (132 cases) were non-selenium supplementation group. Cox proportional hazards models were used to identify the independent predictors for 10-year survival rate of chronic KD. Kaplan-Meier method was used to analyze the 10-year survival rate of patients with chronic KD during the follow-up period and the Log-rank test was used to compare the 10-year survival rate between groups.Results:The follow-up deadline was October 2019. During the follow-up period, a total of 199 patients (199/302, 65.9%) of chronic KD died, including 101 patients (101/170, 59.4%) in the selenium supplementation group and 98 patients (98/132, 74.2%) in the non-selenium supplementation group. In COX proportional hazards model, after adjustment for other baseline characteristics [age, sex, body mass index (BMI), family history of KD, smoking, blood pressure, heart rate, ECG abnormalities, initial cardiothoracic ratio, left ventricular ejection fraction (LVEF), and blood selenium content], selenium supplementation and combined use of angiotensin-converting enzyme inhibitor + β receptor blocker (ACEI + BBs) were protective factors for 10-year survival in patients with chronic KD (selenium supplementation: HR = 0.39, 95% CI: 0.28 - 0.53; ACEI + BBs: HR = 0.57, 95% CI: 0.39 - 0.84). The 10-year survival rate of chronic KD patients after selenium supplementation was significantly higher than that of non-selenium supplementation group (Log-rank test, P < 0.05). Conclusion:Selenium supplementation and combined use of ACEI + BBs in chronic KD patients, are associated with better survival during the 10-year follow-up.

Gene Cloning and Bioinformatics Analysis of Limonene-3-hydroxylase from Schizonepeta tenuifolia / 中国实验方剂学杂志

Objective:This paper aims to clone the cDNA sequence of<italic> limonene</italic>-3-<italic>hydroxylase</italic>（<italic>StL</italic>3<italic>OH</italic>） in <italic>Schizonepeta tenuifolia</italic> and analyze its sequence by bioinformatics. Method:Specific primers were designed based on sequences of<italic> StL</italic>3<italic>OH </italic>gene screened from transcriptome sequencing data of <italic>S. tenuifolia</italic> and the cDNA sequence of <italic>StL</italic>3<italic>OH </italic>gene was cloned by reverse transcription polymerase chain reaction （RT-PCR） and analyzed for its bioinformatics. Result:The <italic>StL3OH</italic> gene cDNA sequence length was 1 598 bp，containing a 1 497 bp long complete open reading frame which encoded 498 amino acids. StL3OH protein had a theoretical relative molecular mass of 56.40 kDa，with a hydrophilic and unstable nature. Bioinformatics analysis showed that StL3OH protein had no signal peptide but had a transmembrane domain which might be located in endoplasmic reticulum. Multiple sequence alignment and cluster analysis showed that the amino acid sequence of MsL3OH protein had a high similarity with StL3OH protein，both of which contained cytochrome P450 heme binding region，belonging to the D subfamily of cytochrome CYP71 family. Codon bias analysis showed that <italic>StL</italic>3<italic>OH</italic> gene preferred guanine/cytosine（G/C） ending codon，with 27 skewed codons， and Nicotiana benthamiana was proven to be the most suitable host for exogenous expression of <italic>StL</italic>3<italic>OH</italic> gene. Conclusion:The cDNA sequence of<italic> StL3OH</italic> gene was cloned from <italic>S. tenuifolia</italic> for the first time，which will provide a basis for further study on the structure and function of StL3OH protein and the regulation mechanism of <italic>StL3OH </italic>gene in the accumulation and biosynthesis of monoterpenes in<italic> S. tenuifolia</italic>.

Nutritional status and behavior change trend of school-age children / 公共卫生与预防医学

Objective To study the nutritional status and behavior of school-age children. Methods In this study, a descriptive study was conducted on 1950 children who underwent physical examination from January to December 2019. There were 955 males, 995 females, 320 7-year-old children, 311 8-year-old children, 333 9-year-old children, 295 10-year-old children, 277 children in 11-year-old group and 414 children in 12-year-old group. Results The malnutrition rate of male was significantly lower than that of female (χ2=14.581, P=0.002); the nutritional status of students of different age groups was statistically significant (χ2=95.265, P=0.000), and the malnutrition status of students aged 10 and 11 was higher than that of other age groups; the difference of picky eating, food response, food preference, drink craving and emotional overeating of students of different ages was found With the increase of age, students' scores of satiety response, slow eating and emotional diet decreased significantly; the differences of over satiety response, slow eating, picky eating, food response, emotional diet reduction and emotional overeating of different genders were significantly higher than those of female patients. Conclusion It is suggested that for school-age children aged 10-11, strengthening nutritional intervention and shaping good eating habits are of positive significance for improving the nutritional status of students.

Desorption electrospray ionization-mass spectrometry in the analysis of natural medicines / 药学学报

Desorption electrospray ionization mass spectrometry (DESI-MS) is a newly emerging in-situ ionization mass spectrometry analysis technology. The ionization process occurs in an open ambient environment at atmospheric pressure, and has the characteristics of simple sample pretreatment, quick and sensitive analysis, and is widely used in biomedicine, pharmaceutical analysis, food safety, environmental monitoring, and material characterization. Natural medicines, such as Chinese herbal medicines, contain a variety of chemical components. Extraction, separation, identification, and in vitro and in vivo efficacy evaluation of natural medicines, especially research on active ingredients with significant efficacy, have received long-term attention. The development of DESI-MS technology provides many new opportunities for direct and rapid analysis of active ingredients in natural medicines. This article briefly introduces the principles, characteristics, influencing factors, and technical progress of DESI-MS technology, and systematically summarizes progress in the research and application of this technology to natural medicines such as Chinese herbal medicines and other plant samples with pharmacological activity. The future application prospects in this field are further presented.

Effect of peroxisome proliferator-activated receptor-γ coactivator-1α on liver injury induced by periodontitis in rats / 华西口腔医学杂志

OBJECTIVES@#To investigate the effect of peroxisome proliferator-activated receptor-γ coactivator-1α (PGC-1α) on liver injury induced by periodontitis in rats.@*METHODS@#Twenty-four male Wistar rats were randomly divided into two groups: control group and periodontitis group, twelve per group. In periodontitis group, the periodontitis models were established for the maxillary first molars in rats by means of "wire ligation+vaccinationwith @*RESULTS@#The probing depth, tooth mobility and sulcus bleeding index in periodontitis group were significantly higher than that in control group. HE staining showed in periodontitis group, hepatic cords ranged disorderly and there were vacuoles in cells and inflammatory cells infiltrated in liver tissues of rats, and there was no obvious abnormality in control group. The qRT-PCR results showed that the mRNA expression levels of @*CONCLUSIONS@#PGC-1α may be involved in the process of periodontitis-induced liver injury in rats.

Study on Sparganium stoloniferum from different regions in China based on appearance characters and ISSR markers / 中国中药杂志

Based on the characteristics and ISSR molecular marker technology, the study is aimed to compare and perform genetic diversity analysis on Sparganium stoloniferum from 7 regions. Molecular identification method was established for S. stoloniferum from Hunan province. Differences among Sparganii Rhizoma samples from seven habitats were analyzed via measuring weight, length, width and thickness of them. Genetic diversity of S. stoloniferum from 7 regions was analyzed by screening out primers amplifying clear band and showing rich polymorphism, then a cultivars dendrogram was built. The target primer was screened out, and the specific band was sequenced. Nine ISSR primers were selected to amplified clear band, rich polymorphism. A total of 73 bands were amplified by nine ISSR primers selected from 27 ISSR primers. On average, each primer produced 8.0 bands. A total of 38 bands were polymorphic, which occupied 52.8% of all bands. The cultivars dendrogram showed the genetic similarity was 0.54-0.94. Genetic similarity coefficient of S. stoloniferum from Jiangsu province, Anhui province and Jiangxi province was big, indicating the differences among them were slight on genetic level. S. stoloniferum from Hunan province is quite different from samples from the other six habitats on appea-rance and genetic level. A specific band(327 bp) in S. stoloniferum from Hunan province was obtained via ISSR-857 primer, and was sequenced. According BLASTn database, there were few sequences similar to the gene fragment and had little correlation with the growth process of plant. ISSR molecular marker technology provides a new idea for the identification of S. stoloniferum. This result confirmed the particularity of S. stoloniferum from ancient Jingzhou.

Influence of short-time pasteurization on essential bioactive components and immune cells in human breast milk / 中华围产医学杂志

Objective:To explore the influence of short-time pasteurization (62.5±0.5℃ for 5 s) on the main bioactive components and immune cells in human breast milk.Methods:Fresh breast milk was collected from 53 women whose premature infants were admitted to the neonatal intensive care unit of the Children's Hospital of Fudan University from May 2020 to October 2020. Each sample (20 ml) was divided into unsterilized, Holder pasteurized (62.5 ℃ for 30 min), or short-time pasteurized groups. The concentration of secretory immunoglobulin A (sIgA), lactoferrin (LTF), lysozyme (LZM), and insulin-like growth factor binding protein-3 (IGFBP-3) in breastmilk whey were detected by enzyme-linked immunosorbent assay and the number of viable immune cells (leukocytes, monocytes, T cells, and B cells) in breastmilk by flow cytometry.Results:(1) A total of 87 breast milk samples were collected. The levels of sIgA, LTF, and LZM were the highest in the unsterilized group, followed by the short-time and Holder pasteurized group [0.42 mg/ml (0.33-0.65 mg/ml) vs 0.40 mg/ml (0.28-0.62 mg/ml) and 0.25 mg/ml (0.17-0.37 mg/ml); (3.57±1.06) vs (3.53±1.11) and (0.85±0.58) mg/ml; 128.60 μg/ml (77.18-203.00 μg/ml) vs 121.70 μg/ml (68.66-188.20 μg/ml) and 83.40 μg/ml (47.40-151.40 μg/ml); all P<0.05]. There was no significant difference in the level of IGFBP-3 among the groups. The median retention rates of sIgA, LTF, and LZM in the Holder pasteurized group were all lower than those in the short-time pasteurized group [55.87% (46.01%-71.41%) vs 96.93% (83.03%-115.90%); 21.72% (12.54%-29.42%) vs 97.88% (88.98%-104.30%); 69.26% (49.42%-89.08%) vs 93.80% (74.85%-110.20%); all P<0.05]. No significant difference in the level of preserved IGFBP-3 was observed between the three groups ( P>0.05). (2) The number of viable leukocytes, monocytes, T cells, and B cells in the Holder pasteurized group were lower than those in the unsterilized group [leukocytes: 185.50 (87.00-356.50) vs 1 271.00 (540.50-2 283.00); monocytes: 12.00 (6.00-16.75) vs 266.00 (137.30-518.80); T cells: 1.00 (0.00-2.00) vs 47.50 (28.50-116.00); B cells: 1.00 (0.00-1.75) vs 21.00(10.00-41.50); all P<0.05]. The percentage of viable leukocyte to the total leukocyte and the viable monocytes, T cells, and B cells to the viable leukocytes were lower in the Holder pasteurized group than those in the unsterilized group [24.80%(16.00%-36.80%) vs 74.20%(63.55%-86.45%); 5.91%(4.09%-8.77%) vs 21.90%(17.40%-29.30%); 0.31%(0.00%-1.31%) vs 4.00%(2.69%-6.43%); 0.30%(0.00%-0.86%) vs 1.27%(0.57%-2.85%); all P<0.05]. A similar trend was observed between short-time pasteurization and unsterilized groups (all P<0.05). (3) The percentages of viable monocytes, T cells, and B cells in their subsets were lower in both Holder and short-time pasteurized groups than those in the unsterilized group [2.94%(1.33%-7.14%) vs 9.72%(5.77%-16.00%) and 52.60%(31.35%-68.75%); 0.00%(0.00%-1.61%) vs 0.49%(0.00%-2.53%) and 28.10%(10.55%-57.00%); 0.00%(0.00%-0.83%) vs 0.24%(0.00%-2.47%) and 13.80%(3.27%-41.00%); all P<0.05].The number and percentage of viable leukocytes in total leukocytes and viable monocytes in total monocytes [leukocytes: 279.50(116.80-548.50), 32.20%(20.70%-45.75%); monocytes: 32.00(21.00- 83.75),15.60%(11.10%-19.15%)] were higher than those in the pasteurized group (all P>0.05). The short-time pasteurized group was noted only for a higher percentage of the viable monocytes to viable leukocytes than the Holder pasteurized group (all P<0.05). Conclusions:Compared with the Holder pasteurization, sIgA, LTF, LZM level, and monocyte activity in breast milk can be better preserved by short-time pasteurization.

Expression profiling and functional verification of flavonoid 3'-hydroxylase gene from leaves of Euryale ferox / 中国中药杂志

Leaves of Euryale ferox are rich in anthocyanins. Anthocyanin synthesis is one of the important branches of the flavonoid synthesis pathway, in which flavonoid 3'-hydroxylase(F3'H) can participate in the formation of important intermediate products of anthocyanin synthesis. According to the data of E. ferox transcriptome, F3'H cDNA sequence was cloned in the leaves of E. ferox and named as EfF3'H. The correlation between EfF3'H gene expression and synthesis of flavonoids was analyzed by a series of bioinforma-tics tools and qRT-PCR. Moreover, the biological function of EfF3'H was verified by the heterologous expression in yeast. Our results showed that EfF3'H comprised a 1 566 bp open reading frame which encoded a hydrophilic transmembrane protein composed of 521 amino acid residues. It was predicted to be located in the plasma membrane. Combined with predictive analysis of conserved domains, this protein belongs to the cytochrome P450(CYP450) superfamily. The qRT-PCR results revealed that the expression level of EfF3'H was significantly different among different cultivars and was highly correlated with the content of related flavonoids in the leaves. Eukaryotic expression studies showed that EfF3'H protein had the biological activity of converting kaempferol to quercetin. In this study, EfF3'H cDNA was cloned from the leaves of E. ferox for the first time, and the biological function of the protein was verified. It provi-ded a scientific basis for further utilizing the leaves of E. ferox and laid a foundation for the further analysis of the biosynthesis pathway of flavonoids in medicinal plants.

A multicenter survey of antibiotic use in very and extremely low birth weight infants in Hunan Province / 中国当代儿科杂志

OBJECTIVE@#To investigate the current status of antibiotic use for very and extremely low birth weight (VLBW/ELBW) infants in neonatal intensive care units (NICUs) of Hunan Province.@*METHODS@#The use of antibiotics was investigated in multiple level 3 NICUs of Hunan Province for VLBW and ELBW infants born between January, 2017 and December, 2017.@*RESULTS@#The clinical data of 1 442 VLBW/ELBW infants were collected from 24 NICUs in 2017. The median antibiotic use duration was 17 days (range: 0-86 days), accounting for 53.0% of the total length of hospital stay. The highest duration of antibiotic use was up to 91.4% of the total length of hospital stay, with the lowest at 14.6%. In 16 out of 24 NICUs, the antibiotic use duration was accounted for more than 50.0% of the hospitalization days. There were 113 cases with positive bacterial culture grown in blood or cerebrospinal fluid, making the positive rate of overall bacterial culture as 7.84%. The positive rate of bacterial culture in different NICUs was significantly different from 0% to 14.9%. The common isolated bacterial pathogens Klebsiella pneumoniae was 29 cases (25.7%); Escherichia coli 12 cases (10.6%); Staphylococcus aureus 3 cases (2.7%). The most commonly used antibiotics were third-generation of cephalosporins, accounting for 41.00% of the total antibiotics, followed by penicillins, accounting for 32.10%, and followed by carbapenems, accounting for 13.15%. The proportion of antibiotic use time was negatively correlated with birth weight Z-score and the change in weight Z-score between birth and hospital discharge (r=-0.095, -0.151 respectively, P<0.01), positively correlated with death/withdrawal of care (r=0.196, P<0.01).@*CONCLUSIONS@#Antibiotics used for VLBW/ELBW infants in NICUs of Hunan Province are obviously prolonged in many NICUs. The proportion of routine use of third-generation of cephalosporins and carbapenems antibiotics is high among the NICUs.

An investigation of Kaschin-Beck disease in Shaanxi Province in 2018 / 中华地方病学杂志

Objective To master the basic conditions of Kaschin-Beck disease areas and patients in Shaanxi Province,and provide a theoretical basis for prevention and control of Kaschin-Beck disease.Methods In 2018,the basic situation of the villages in Kaschin-Beck disease areas of Shaanxi Province and the implementation of prevention and control measures were investigated.The data of general population information,condition and treatment were collected from all the disease affected villages.Results The Kaschin-Beck disease areas of Shaanxi Province involves 4 194 disease affected villages in 62 counties.Among them,3 886 villages had completed the whole village water reform,215 villages changed some of the water,and 93 villages had not changed water;there were 3 835 disease affected villages met the hygienic standard of drinking water,accounting for 91.44％.A total of 60 157 patients with Kaschin-Beck disease were diagnosed.The male-female ratio was 1.0:0.9 (31 531:28 626).The age of the patients was mainly between 40 and 65 years old.There were no new cases of children.Among them,38 448 were degree Ⅰ patients,18 589 were degree Ⅱ patients and 3 120 were degree Ⅲ patients.The occupational distribution of patients was mainly peasant,with 59 703 cases,accounting for 99.25％.The distribution of education was mainly primary school,with 31 108 cases,accounting for 51.71％.There were 876 patients who had completed surgery and 23 284 patients who had been treated with long-term medication.Conclusions In recent years,there is no new case of Kaschin-Beck disease in children in Shaanxi Province.However,the condition of Kaschin-Beck disease in adults is serious,and great efforts should be made in rescue.

Dynamic changes of lung function in mice with pulmonary fibrosis / 医学研究生学报

Objective　Pulmonary function testing is a commonly used indicator for clinical evaluation of the degree of pulmonary fibrosis in patients. This paper aims to investigate the dynamic changes of lung function in mice with pulmonary fibrosis and to establish a range of reference values for lung function parameters in normal Kunming mice.　Methods　Twenty-eight SPF Kunming mice were randomly divided into normal control group (n=14) and model group (n=14). After anesthesia with 10% chloral hydrate, the normal control group only underwent tracheal puncture. The model group received intratracheal puncture and injection of bleomycin (BLM) (5 mg/kg body weight), and the lung function indicators of all mice were detected in the same order on the 1st, 2nd, 3rd and 4th weekends after modeling: Ti, Te, PIF, PEF, TV, EV, RT, MV, f, Penh and EF50.　Results　After intratracheal BLM injection, mice in the model group showed decreased hair softness and smoothness, hair loss and decreased activity after the 2nd week. Compared with the control group, Ti, Te and RT values in the model group significantly increased at week 4 (P<0.05), while the values of PEF, RT, MV, f and EF50 decreased significantly at the same week (P<0.05). Compared with the model group at week 1, the differences in Ti, Te, RT and f values at week 2, 3 and 4 were statistically significant (P<0.05); the differences in MV and EF50 values at week 3 and 4 were statistically significant (P<0.05); while the PIF values only showed differences at week 4 (P<0.001). Compared with the Penh values in the control group at week 2, 3 and 4 (0.553±0.189, 0.662±0.164, 0.712±0.189), the differences of the model group (0.820±0.205, 0.936±0.188, 1.053±0.236) showed statistical significance (P<0.001). Compared with the model group at week 1, the differences of Penh values in the model group only showed statistical significance at week 3 and 4 (P<0.05). Through four-week lung function test, various parameters were obtained, among which the normal range of the main index Penh value was 0.27-0.88.　Conclusion　The lung function detected by the non-invasive whole body plethysmography system was stable and reliable with good effects; the lung function in mice with the BLM-induced pulmonary fibrosis continued to decrease within four weeks. Penh, which reflects airway resistance, can be used for overall screening of the lung function among the test mice after two weeks of modeling.

Establishment of Ace2 knockout mouse model with CRISPR/Cas9 gene targeting technology / 生理学报

The aim of the study was to establish Ace2 (angiotensin-converting enzyme 2) knockout mouse model with CRISPR/Cas9 gene targeting technology. A vector targeting Ace2 gene knockout was constructed with the primers of single-guide RNA (gRNA), and then transcribed gRNA/Cas9 mRNA was micro-injected into the mouse zygote. The deletion of exons 3 to 18 of Ace2 gene in mice was detected and identified by PCR and gene sequencing. The Ace2 gene knock-out mice were bred and copulated. Ace2 protein and mRNA expression were detected by Western blot and qRT-PCR in F3 progeny knock-out male mice. The gRNA expression vector was successfully constructed and transcribed in vitro, and active gRNA and Cas9 mRNA were injected directly into zygote. The deletion of exons 3 to 18 of Ace2 gene in six positive founder mice as the F0 generation were confirmed by PCR and gene sequencing. Six founder mice were mated with wild-type mice, then achieved F1 generation were mated and produced F2 generation. The female positive mouse of F2 was selected to mate with wild-type mice and produce Ace2 mice of F3 generation. Ace2 mRNA and protein were not detected in tissues of these Ace2 mice. In conclusion, a mouse model with Ace2 deficiency has been successfully established with CRISPR/Cas9 technique, which shall lay a foundation for future investigation of Ace2.