BACKGROUND:It is urgent to improve the study on the molecular mechanism of Ban's Culuan Zhuyun Decoction improving oocyte quality in polycystic ovary syndrome.OBJECTIVE:To observe the effects of Ban's Culuan Zhuyun Decoction on oocyte quality in a mouse model of polycystic ovary syndrome and to explore the underlying mechanisms of its intervention in polycystic ovary syndrome.METHODS:Subcutaneous injection of dehydroepiandrosterone sulfate was used to establish the polycystic ovary syndrome model in 21-day-old female Kunming mice,and the treatment was conducted for 21 consecutive days.The estrous cycle and pregnancy was recorded.ELISA was used to detect serum sex hormone levels.The rate of apoptosis in oocytes was detected using Annexin V staining.The level of reactive oxygen species in oocytes was detected using dichlorodihydrofluorescein diacetate.The condition of spindle bodies and chromosomes in oocytes were detected using the immunofluorescence method.Network pharmacology and molecular docking were used to verify the binding properties of Ban's Culuan Zhuyun Decoction core active components and oocyte maturation-related factors(growth differentiation factor 9 and bone morphogenetic protein 15).Real-time fluorescence quantitative PCR and western blot were used to detect the mRNA and protein expression levels of growth differentiation factor 9 and bone morphogenetic protein 15 in oocytes,respectively.RESULTS AND CONCLUSION:(1)Ban's Culuan Zhuyun Decoction core active components(quercetin,kaempferol,and β-sitosterol)showed good binding activities with growth differentiation factor 9 and bone morphogenetic protein 15.(2)Ban's Culuan Zhuyun Decoction ameliorated the estrous cycle,regulated serum hormone,increased the pregnancy,decreased the rate of apoptosis,declined the level of reactive oxygen species,diminished the rate of abnormal spindle assembly and chromosome loss(P<0.01,P<0.05);and promoted the mRNA and protein expression of growth differentiation factor 9 and bone morphogenetic protein 15(P<0.05).Therefore,Ban's Culuan Zhuyun Decoction may improve the oocyte quality and increase the fertility of polycystic ovary syndrome mice by regulating the gene expression of growth differentiation factor 9 and bone morphogenetic protein 15.

BACKGROUND:It is urgent to improve the study on the molecular mechanism of Ban's Culuan Zhuyun Decoction improving oocyte quality in polycystic ovary syndrome.OBJECTIVE:To observe the effects of Ban's Culuan Zhuyun Decoction on oocyte quality in a mouse model of polycystic ovary syndrome and to explore the underlying mechanisms of its intervention in polycystic ovary syndrome.METHODS:Subcutaneous injection of dehydroepiandrosterone sulfate was used to establish the polycystic ovary syndrome model in 21-day-old female Kunming mice,and the treatment was conducted for 21 consecutive days.The estrous cycle and pregnancy was recorded.ELISA was used to detect serum sex hormone levels.The rate of apoptosis in oocytes was detected using Annexin V staining.The level of reactive oxygen species in oocytes was detected using dichlorodihydrofluorescein diacetate.The condition of spindle bodies and chromosomes in oocytes were detected using the immunofluorescence method.Network pharmacology and molecular docking were used to verify the binding properties of Ban's Culuan Zhuyun Decoction core active components and oocyte maturation-related factors(growth differentiation factor 9 and bone morphogenetic protein 15).Real-time fluorescence quantitative PCR and western blot were used to detect the mRNA and protein expression levels of growth differentiation factor 9 and bone morphogenetic protein 15 in oocytes,respectively.RESULTS AND CONCLUSION:(1)Ban's Culuan Zhuyun Decoction core active components(quercetin,kaempferol,and β-sitosterol)showed good binding activities with growth differentiation factor 9 and bone morphogenetic protein 15.(2)Ban's Culuan Zhuyun Decoction ameliorated the estrous cycle,regulated serum hormone,increased the pregnancy,decreased the rate of apoptosis,declined the level of reactive oxygen species,diminished the rate of abnormal spindle assembly and chromosome loss(P<0.01,P<0.05);and promoted the mRNA and protein expression of growth differentiation factor 9 and bone morphogenetic protein 15(P<0.05).Therefore,Ban's Culuan Zhuyun Decoction may improve the oocyte quality and increase the fertility of polycystic ovary syndrome mice by regulating the gene expression of growth differentiation factor 9 and bone morphogenetic protein 15.

Pheretima has a long history of medication, its original name was earthworm, and it was first recorded in Shennong Bencaojing, which was listed as inferior product. In the 2015 edition of Chinese Pharmacopoeia, two varieties of Pheretima were included. However, due to the variety of Pheretima, there are mixed non-pharmacopoeia collection of Pheretima family status. Based on the systematic review of ancient and modern literature, this paper conducts herbal textual research on Pheretima in terms of name, origin, distribution of origin, genuine production area, harvesting time, processing methods, etc. Based on the analysis of various ancient books and modern research documents of herbal medicine and their accompanying drawings, it is found that the Pheretima and white-necked Pheretima mentioned in ancient books are the general names of the genus Pheretima. The ancient people thought that white-necked Pheretima was a good medicinal material, which was the same as the opinion that Guangdilong was better in quality than Tudilong in modern research. In ancient and modern literature, the origin of earthworm is relatively consistent, but due to the change of environment, the output of wild Pheretima is reduced, and now the output of Pheretima is mainly artificial breeding. In ancient times, harvesting should be as far as possible in spring, summer and autumn. However, in modern times, the best harvest time is autumn. Different processing methods of earthworm in different ages and regions are different. Attention should be paid to following and inheriting the ancient processing methods, combining with modern research techniques, the quantitative standard of processing of Pheretima should be formulated, so that Pheretima medicinal materials can be applied comprehensively and effectively. This research provides the basis for the original source, resource development, correct use, genuine producing area and processing method determination of Pheretima.

Objective:To investigate the risk factors of Crohn′s disease (CD)-related gastrointestinal stenosis, and to summarize and analyze the corresponding treatments.Methods:From January 2010 to December 2018, 122 patients diagnosed with CD and hospitalized in the Seventh Medical Center, PLA General Hospital were selected including 72 patients in gastrointestinal stenosis group and 50 patients in non-gastrointestinal stenosis group. The gender, age of onset, course of disease, location of lesions involved (Montreal classification), disease activity, extraintestinal manifestations, application of therapeutic drugs, and complications were compared between the two groups. The treatment of CD patients with gastrointestinal stenosis was analyzed. Multivariate logistic regression was used to analyze the risk factors of CD patients with gastrointestinal stenosis. The independent sample t test, Mann-Whitney U test and chi-square test were used for statistical analysis. Results:The age of onset of patients in gastrointestinal stenosis group was older than that in non-gastrointestinal stenosis group ((37.6±15.1) years old vs. (30.8±14.7) years old), and course of disease was longer than that of non-gastrointestinal stenosis group (72 months, 11 to 492 months vs. 45 months, 3 to 240 months); and the differences were statistically significant ( t=-2.044, Z=-2.770; P=0.018, 0.006). The proportion of patients with ileum involvement of the gastrointestinal stenosis group was lower than that of the non-gastrointestinal stenosis group (69.4%, 50/72 vs. 86.0%, 43/50), and the proportion of severe patients was higher than that of the non-gastrointestinal stenosis group (15.3%, 11/72 vs. 4.0%, 2/50); and the differences were statistically significant ( χ2=4.463 and 3.942, P=0.035 and 0.047). There were no significant differences in gender, use of therapeutic drugs, extraintestinal manifestations, application of therapeutic drugs or the incidence of complications between the patients of two groups (all P>0.05). The results of multivariate logistic regression showed that the age of onset and course of disease were risk factors of CD-related gastrointestinal stenosis ( β=0.028, odds ratio ( OR)=1.028, 95% confidence interval ( CI) 1.000 to 1.056, P=0.046; β=0.008, OR=1.008, 95% CI 1.002 to 1.015, P=0.013). Further stratified analysis revealed that the incidence rates of CD-related gastrointestinal stenosis in patients with age of onset over 40 years old and course of disease more than five years were higher than those of patients with age of onset less than 40 years old and course of disease less than five years (76.3%, 29/38 vs. 51.2%, 43/84; 68.4%, 39/57 vs. 50.8%, 33/65), and the differences were statistically significant ( OR=3.072, 95% CI 1.298 to 7.272, P=0.009; OR=2.101, 95% CI 1.002 to 4.406, P=0.048). Among the 72 CD patients with gastrointestinal stenosis, 15 cases (20.8%) were treated with medicine and nutrition, without endoscopic or surgical treatment. Fifty-two patients (72.2%) underwent surgical treatment, among them six patients (11.5%) received twice surgery, the interval between the two operations was 46 months (1 to 204 months), and eight patients (15.4%) had postoperative complications. Twenty-one patients (29.2%) were treated with endoscopic dilatation, and no complications occurred after surgery. Five patients (23.8%) underwent surgical treatment during the follow-up period. Conclusions:The age of onset over 40 years old and the course of disease more than five years are the risk factors of CD-related gastrointestinal stenosis. Individualized medical treatment is the basis for the treatment of CD-related gastrointestinal stenosis. Surgery is still the main treatment. The endoscopic treatment is safety and can delay or avoid surgery to a certain extent.

Objective: To establish a method for determining the content of total polysaccharides in decoction pieces of Polyporus,analyze the content of total polysaccharides in samples with different sources and grades. Method: The relative molecular weight and the polydispersity index of polysaccharides in decoction pieces of Polyporus were measured by a high performance gel chromatography coupled with a multi-angle laser light scattering and refractive index system.Dextran with similar molecular weight as polysaccharides was selected as the reference substance.Orthogonal experiment and single factor tests were used to optimize the pretreatment conditions for the determination of total polysaccharides in Polyporus.Polysaccharides in Polyporus with different areas and grades were determined by anthrone-sulfuric acid colorimetric method at 630 nm. Result: The linearity,stability,precision,repeatability and recovery rate of the established method all reached the standards,respectively.The content of total polysaccharides in samples from different areas ranged from 0.87% to 1.39%.The content of total polysaccharides in samples with different grades was 1.40% for first-grade pieces,1.21% for second-grade pieces, and 1.03% for third-grade pieces. Conclusion: The established method is simple,accurate and reproducible,and it can be used for the determination of polysaccharides in decoction pieces of Polyporus.The content of polysaccharides in samples from different origins varies greatly.The content of polysaccharides in samples with different grades shows a certain regularity.The content of polysaccharides is the highest in the first-grade pieces,followed by the content in the second-grade,and the lowest in the third-grade.The results can provide a reference for formulating limits for the content of total polysaccharides and the grade standard of decoction pieces of Polyporus.

The grading and quality analysis methods for different commercial Glycyrrhiza Polygalae Radix slices were established. The qualities of different grade samples were analyzed and compared, in order to provide useful information for the formulation of the grading standards of Glycyrrhiza Polygalae Radix slices. A total of 34 batches of Glycyrrhiza Polygalae Radix slice samples collected from 12 companies were divided into two grades: first-grade (diameter ≥ 3.0 mm) and second-grade (diameter < 3.0 mm). Thin-layer chromatography (TLC), multi-component content determination and fingerprint analysis were used to analyze the qualities of different grades of Glycyrrhiza Polygalae Radix slices, and the fingerprints were statistically analyzed using partial least squares-discriminant analysis (PLS-DA) and orthogonal partial least squares-discriminant analysis (OPLS-DA). The results showed that the established TLC method can simultaneously identify three major types of components, including sugar esters, xanthones, and saponins in Glycyrrhiza Polygalae Radix slices, and has obvious advantage compared to the existing methods for its rich information, low cost, and easy or safe operation. The multi-component determination showed that the contents of three index components (polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose and tenuifolin) in the first-grade products of Glycyrrhiza Polygalae Radix slices were lower than those in the second-grade products. The results of PLS-DA and OPLS-DA indicated significant differences were observed between the first-grade and second-grade products, with sibiricose A5, sibiricose A6, polygalaxanthone Ⅲ, 3,6'-disinapoyl sucrose and tenuifoliside A being identifies as the major differentiate markers.

Objective To study the effects of cystic fibrosis transmembrane conductance regulator (CFTR) on neonatal rats with bronchopulmonary dysplasia (BPD).Method The hyperoxia (FiO2＞ 90％)-induced neonatal BPD rat models were established and assigned into three groups:the model group,the agonist group and the antagonist group.Room air (FiO2 21％) was inhaled by the rats in the control group.50 μl of phosphate buffered saline (PBS),genistein (50 mg/kg),arachidonic acid (500 mg/kg) and PBS were injected intraperitoneally respectively in the model group,the agonist group,the antagonist group and the control group at 24,48 and 72 h after birth.The survival rates of the neonatal rats were calculated,the survival curves were drawn,the pathological changes of the lung tissues were examined (the control group and the model group:3,14 and 21 d after birth;the agonist group and antagonist group:14 and 21 d after birth),and the expression of CFTR were studied using western blot method.The acute lung injury scores of the model group,the agonist group and the antagonist group were compared and the gray value was analyzed using Graphpad software.Result (1) The survival rates in the control group,the model group,the agonist group and the antagonist group were 96.8％,93.3％,100％ and 34.5％ respectively.The antagonist group had significantly lower survival rate than the other three groups (P＜0.001).(2)The alveoli developed gradually with age in the control group.The pulmonary pathology of the model group showed:alveolar congestion,hemorrhage,infiltration or aggregation of neutrophils in airspace or vessel wall,thickness of alveolar wall,with some enlarged alveolar spaces and reduced alveolar cavities.As the inflammation gradually decreased,some alveolar spaces significantly enlarged and the numbers of alveolar cavities significantly reduced.No significant differences existed of the acute lung injury scores among the agonist group,the antagonist group and the model group at 14 and 21 d after birth (P＞0.05).(3) The expressions of CFTR in the lungs were lower in the model group than the control group 3 d after birth (P＜0.01).No significant differences existed of the CFTR expression between the model group and the control group 14 d after birth(P＞0.05).The CFTR expression was much higher in the agonist group than the model group (P＜0.01) and also higher in the antagonist group than the model group (P＜0.05) 14 d after birth.The CFTR expression was lower in the model group than the control group,and higher in the agonist group than the model group 21 d after birth (P＜ 0.05).No significant differences existed of CFTR expression between the antagonist group and the model group 21 d after birth (P＞0.05).Conclusion CFTR may play a protective role in the pathogenesis of BPD.

Salt is an important excipient for Chinese medicine salt. The standardization of its quality is conducive to ensuring the quality of Chinese medicine pieces. In this paper, the processing with brine and the processing auxiliary salt were studied from three aspects： the history evolution of salt, the history evolution of processing with brine, and the modern research of processing with brine. It has been found that salt plays an important role in the history of China. It has a long history and a wide variety. The salt used in the processing of traditional Chinese medicine mainly includes three categories： common salt, halitum and white salt. The quality of salt is closely related to its origin and processing, mainly based on the color and the place of origin. In ancient times, the varieties of salt used in the production of different Chinese herbal medicines were different, which might be related to the nature of drugs. The primary purpose of processing with brine is to increase the efficacy of drugs. At present, there are many reports on optimizing the preparation technology of processing with brine, but the evaluation indexes are quite different, and its scientific nature is to be discussed. The processing method with brine and its processing auxiliary materials are lacking of relevant evaluation standards and quality standards, which is not conducive to the healthy development of Chinese herbal pieces. In this paper, the relevant literature was studied in order to provide reference for the establishment of standards for salt processing excipient in traditional Chinese medicine.

PurposeTo observe the effects of rhIGF-1 on kidney in diabetic rats. MethodsUsing biochemistry, radioimmunoassay, molecular biology (RT-PCR). Results(1) 24 h UAER,24 h uriary volume in rhIGF-1 group is lower than that of diabetic control group; (2) The level of sertan IGF-1 ,kidney IGF-1 and IGF-1 mRNA in diabetic control group is lower than that of normal control group;The level of serum IGF-1 in rhIGF-1 group is higher than that of diabetic control group;no differences were found in the levels of kidney IGF-1 and kidney IGF-1 mRNA between diabetic control group and rhIGF-1 group; (3) The level of serum GH in diabetic control group is higher than that of normal control group; The level of serum GH in rhIGF-1 group is lower than that of diabetic control group; (4) rhIGF-1 might have some protective effects on diabetic nephropathy via electron microscope. ConclusionsrhIGF-1 doesn't increase the damage of diabetic kidney tissue.

The changes of parathyroid hormone (PTH),calcitonin(CT)of three different anesthesia methods were observed perioperatively in 38 patients who were divided into two age groups (22 ~ 50 and 51 ~ 60). Postoperative serum Ca, P levels decreased as compared with preoperative values,espe.cially in the old. age groups with no statistic significance. These results indicate that during anesthesia and operative stress the CDS is inhibited and the regulation of the autonomic nervous system is affected, with obscure the relationship between parathyroid hormone, calcitonin and Ca, P with the alteration of Ca,P metabolism. Serum Ca may increase to the baseline by supplement of calcium gluconate during and after operation, but the operation after 4 ~ 5 days, serum Ca may also return to normal without any intervention.