Manufacturing classification system for oral solid dosage forms of traditional Chinese medicines (Ⅱ): classification of tablets disintegration behavior / 中国中药杂志

In this paper, 50 batches of representative traditional Chinese medicine tablets were selected and the disintegration time was examined with the method in Chinese Pharmacopoeia. The disintegration time and disintegration phenomenon were recorded, and the dissolution behaviors of water-soluble and ultraviolet-absorbent components during the disintegration process of tablets were characterized by self-control method. The results revealed that coating type and raw material type influenced the disintegration time of tablets. It was found that only 4% of traditional Chinese medicine tablets had obvious fragmentation during the disintegration process, while 96% of traditional Chinese medicine tablets showed gradual dissolution or dispersion. Furthermore, according to the disintegration speed, disintegration phenomenon, and whether the cumulative dissolution of measured components was > 90% at complete disintegration, a disintegration behavior classification system(DBCS) was created for the regular-release traditional Chinese medicine tablets. As a result, the disintegration behaviors of 50 batches of traditional Chinese medicine tablets were classified into four categories, i.e. ⅠA_2, ⅠB_1, ⅡB_1, and ⅡB_2. traditional Chinese medicine tablets(Class I) with disintegration time ≤ 30 min were defined to be rapid in disintegration, which can be the objective of optimization or improvement of Chinese herbal extract(semi extract) tablets. Different drug release models were used to fit the dissolution curve of traditional Chinese medicine tablets with gradual dissolution or dispersion phenomenon(i.e. Type B tablets). The results showed that the dissolution curves of water-soluble components in the disintegration process conformed to the zero order kinetics and the Ritger-Peppas model. It could be inferred that the disintegration mechanisms of type B tablets were a combination of dissolution controlled and swelling controlled mechanisms. This study contributes to understanding the disintegration behavior of traditional Chinese medicine tablets, and provides a reference for the design and improvement of disintegration performance of traditional Chinese medicine tablets.

Electroacupuncture Attenuated Phenotype Transformation of Vascular Smooth Muscle Cells via PI3K/Akt and MAPK Signaling Pathways in Spontaneous Hypertensive Rats / 中国结合医学杂志

OBJECTIVE@#To investigate whether the antihypertensive mechanism of electroacupuncture (EA) is associated with attenuating phenotype transformation of vascular smooth muscle cells (VSMCs) via phosphoinositide3-kinase (PI3K)/protein kinase B (Akt) and mitogen-activated protein kinase (MAPK) signaling pathways.@*METHODS@#Eight Wistar-ktoyo (WKY) rats were set as normal blood pressure group (normal group). A total of 32 spontaneous hypertensive rats (SHRs) were randomly divided into 4 groups using random number tables: a model group, an EA group, an EA+PI3K antagonist group (EA+P group), and an EA+p38 MAPK agonist+extracellular signal-regulated kinase (ERK) agonist group (EA+M group) (n=8/group). SHRs in EA group, EA+P group and EA+M group received EA treatment 5 sessions per week for continuous 4 weeks, while rats in the normal and model groups were bundled in same condition. The systolic blood pressure (SBP), diastolic blood pressure (DBP), and mean arterial pressure (MAP) of each rat was measured at 0 week and the 4th week. After 4-week intervention, thoracic aorta was collected for hematoxylin-eosin (HE) staining, immunohistochemistry [the contractile markers α-smooth muscle actin (α-SMA) and calponin and the synthetic marker osteopontin (OPN)] and Western blot [α-SMA, calponin, OPN, PI3K, phosphorylated-Akt (p-Akt), Akt, p-p42/44 ERK, total p42/44 ERK, p-p38 MAPK and total p38 MAPK].@*RESULTS@#EA significantly reduced SBP, DBP and MAP (P<0.01). HE staining showed that the wall thickness of thoracic aorta in EA group was significantly decreased (P<0.01). From results of immunohistochemistry and Western blot, EA increased the expression of α-SMA and calponin, and decreased the expression of OPN (P<0.01). In addition, the expression of PI3K and p-Akt increased (P<0.01), while the expression of p-p42/44 ERK and p-p38 MAPK decreased in EA group (P<0.01). However, these effects were reversed by PI3K antagonist, p38 MAPK agonist and ERK agonist.@*CONCLUSIONS@#EA was an effective treatment for BP management. The antihypertensive effect of EA may be related with inhibition of phenotypic transformation of VSMCs, in which the activation of PI3K/Akt and the repression of MAPK pathway were involved.

Influence of electroacupuncture on ghrelin and the phosphoinositide 3-kinase/protein kinase B/endothelial nitric oxide synthase signaling pathway in spontaneously hypertensive rats / 中西医结合学报

OBJECTIVE@#To investigate the influence of electroacupuncture (EA) on ghrelin and the phosphoinositide 3-kinase/protein kinase B/endothelial nitric oxide synthase (PI3K/Akt/eNOS) signaling pathway in spontaneously hypertensive rats (SHRs).@*METHODS@#Eight Wistar-Kyoto rats were used as the healthy blood pressure (BP) control (normal group), and 32 SHRs were randomized into model group, EA group, EA plus ghrelin group (EA + G group), and EA plus PF04628935 group (a potent ghrelin receptor blocker; EA + P group) using a random number table. Rats in the normal group and model group did not receive treatment, but were immobilized for 20 min per day, 5 times a week, for 4 continuous weeks. SHRs in the EA group, EA + G group and EA + P group were immobilized and given EA treatment in 20 min sessions, 5 times per week, for 4 weeks. Additionally, 1 h before EA, SHRs in the EA + G group and EA + P group were intraperitoneally injected with ghrelin or PF04628935, respectively, for 4 weeks. The tail-cuff method was used to measure BP. After the 4-week intervention, the rats were sacrificed by cervical dislocation, and pathological morphology of the abdominal aorta was observed using hematoxylin-eosin (HE) staining. Enzyme-linked immunosorbent assay (ELISA) was used to detect the levels of ghrelin, nitric oxide (NO), endothelin-1 (ET-1) and thromboxane A2 (TXA2) in the serum. Isolated thoracic aortic ring experiment was performed to evaluate vasorelaxation. Western blot was used to measure the expression of PI3K, Akt, phosphorylated Akt (p-Akt) and eNOS proteins in the abdominal aorta. Further, quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to measure the relative levels of mRNA expression for PI3K, Akt and eNOS in the abdominal aorta.@*RESULTS@#EA significantly reduced the systolic BP (SBP) and diastolic BP (DBP) (P < 0.05). HE staining showed that EA improved the morphology of the vascular endothelium to some extent. Results of ELISA indicated that higher concentrations of ghrelin and NO, and lower concentrations of ET-1 and TXA2 were presented in the EA group (P < 0.05). The isolated thoracic aortic ring experiment demonstrated that the vasodilation capacity of the thoracic aorta increased in the EA group. Results of Western blot and qRT-PCR showed that EA increased the abundance of PI3K, p-Akt/Akt and eNOS proteins, as well as expression levels of PI3K, Akt and eNOS mRNAs (P < 0.05). In the EA + G group, SBP and DBP decreased (P < 0.05), ghrelin concentrations increased (P < 0.05), and the concentrations of ET-1 and TXA2 decreased (P < 0.05), relative to the EA group. In addition, the levels of PI3K and eNOS proteins, the p-Akt/Akt ratio, and the expression of PI3K, Akt and eNOS mRNAs increased significantly in the EA + G group (P < 0.05), while PF04628935 reversed these effects.@*CONCLUSION@#EA effectively reduced BP and protected the vascular endothelium, and these effects may be linked to promoting the release of ghrelin and activation of the PI3K/Akt/eNOS signaling pathway.

Recent research on the treatment of spinal muscular atrophy / 中国当代儿科杂志

Spinal muscular atrophy (SMA) is an autosomal recessive neuromuscular disease characterized by progressive muscular weakness and atrophy. SMA, as an inherited disease, is the leading cause of death in infants and young children. Rapid progress has been made in the research field of SMA in recent years, and some related treatment drugs have been successfully approved for marketing. This article reviews the recent research advances in the treatment of SMA.

Analgesic Effect and Action Mechanisms of Wenjing Zhitong Prescription on Primary Dysmenorrhea / 中国实验方剂学杂志

Objective:To evaluate the analgesic effects of Wenjing Zhitong prescription （WZP） and explore its possible analgesic mechanisms so as to provide experimental basis for research and development of new Chinese medicine. Method:Analgesic effects of WZP were evaluated by observing the writhing latency and number in the writhing models which were induced by oxytocin in rats as well as those induced by acetic acid and prostaglandin E₁ （PGE₁）， respectively in mice. Effect of WZP on uterine contraction frequency， amplitude and activity were evaluated by observing the oxytocin-induced contraction of uterine smooth muscle in rats and rabbits <italic>in vivo</italic>. In the oxytocin-induced rat writhing models， the content of prostaglandin F₂<italic>_α</italic>（PGF₂<italic>_α</italic>） and prostaglandin E₂（PGE₂） in rat uterine tissues and the content of beta-endorphins （<italic>β</italic>-EP） in rat serum were detected by enzyme-linked immunosorbent assay （ELISA）. Expression of estrogen receptor （ER） and oxytocin receptor （OTR） in rat uterine were tested by Real-time polymerase chain reaction（Real-time PCR） method to investigate the possible molecular mechanism of WZP for its analgesic effect. Result:Results of analgesic effect showed that in oxytocin-induced rat writhing experiment， the number of writhing responses in both the WZP （1.5，3.0 g·kg^-1） group was lower than than in the model group （<italic>P</italic><0.05）. In acetic acid-induced mice writhing experiment， the latency of writhing response in WZP （6.0 g·kg^-1） group was significantly prolonged as compared with that in model group <italic>（P</italic><0.01）， and the number of writhing response was significantly reduced （<italic>P</italic><0.05）. In PGE₁-induced mice writhing model， the writhing number in WZP （1.5，3.0，6.0 g·kg^-1） groups was significantly lower than that in the model group （<italic>P</italic><0.05，<italic>P</italic><0.01）. Results of effect on uterine smooth muscle demonstrated that WZP （0.38，0.75，1.50 g·kg^-1） could significantly reduce the frequency of uterine smooth muscle contraction in rabbits （<italic>P</italic><0.05，<italic>P</italic><0.01）， WZP （0.75，1.50，3.00 g·kg^-1） could significantly reduce the contractile amplitude and activity of smooth muscle in the uterus of rats （<italic>P</italic><0.05）. Results of molecular mechanisms of analgesic effects showed that the WZP （0.75，1.50，3.00 g·kg^-1） significantly reduced the content of PGF₂<italic>_α</italic> and the ratio of PGF₂<italic>_α</italic> to PGE₂ in the uterine tissue of rats （<italic>P</italic><0.01）. In the WZP （3.00 g·kg^-1） group， the levels of <italic>β</italic>-EP in the serum of rats were significantly increased （<italic>P</italic><0.01）， and the levels of OTR in uterus of rats in the WZP （1.50，3.00 g·kg^-1） group were significantly decreased （<italic>P</italic><0.05）. Conclusion:Pharmacological studies demonstrated potent analgesic effect of WZP， and such analgesic effect were mediated by significantly inhibiting contraction of uterine smooth muscle， decreasing the contents of PGF₂<italic>_α </italic>and ratio of PGF₂<italic>_α</italic>/PGE₂， reducing OTR expression in uterine as well as increasing the amount of <italic>β</italic>-EP in serum.

Evaluation of taste changes of Scutellariae Radix before and after wine-frying based on electronic ongue technology and its application in identification of Scutellariae Radix pieces / 中国中药杂志

Scutellariae Radix(Huangqin) is a well-known traditional Chinese medicine(TCM) used for the treatment of clearing heat in clinical application. It is bitter-cold by using directly, but the bitter-cold property can be relieved after wine-frying. The study of taste changes before and after wine-frying of Scutellariae Radix is of great significance in identifying Scutellariae Radix and wine-processed Scutellariae Radix and clarifying the traditional theory of wine-processing. In this experiment, 10 batches of Scutellariae Radix and wine-processed Scutellariae Radix were prepared. The contents of 5 flavonoids were determined by high performance liquid chromatography(HPLC), and principal component analysis(PCA) was performed with 5 flavonoids as variables. As a result, the contents were different in different batches of Scutellariae Radix, but Scutellariae Radix and wine-processed Scutellariae Radix could not be distinguished. Five sensory attributes(sour, salty, fresh, sweet, and bitter) were evaluated by artificial tasting, and the response values of 7 sensors(AHS, AHS, PKS, CTS, NMS, CPS, ANS, SCS) representing the taste of pieces were detected by electronic tongue. The correlation between sensory evaluation and response values of the electronic tongue were analyzed, and the results showed that the sensory evaluation of sour, salty, fresh, sweet, bitter and AHS, CTS, NMS, ANS, SCS sensors had different degrees of correlation, indicating that the electronic tongue technology can be used as an alternative to artificial taste and can serve as a means for quantifying the taste, and it can be used to evaluate the taste of TCM pieces. The taste method was used to analyze the response values of the electronic tongue, and the results showed that the bitterness of wine-processed Scutellariae Radix decreased and the salty taste increased. PCA was used to analyze taste changes before and after wine-processed Scutellariae Radix, and the results showed that taste differences between 2 pieces were divided into 2 categories. PCA loading scattering plots showed that response of saltiness and bitterness were the major factors to affect overall taste in Scutellariae Radix and wine-processed Scutellariae Radix. Based on electronic tongue response values, the Fisher discriminant model for Scutellariae Radix and wine-processed Scutellariae Radix was established, which showed that it could effectively discriminate them with a recognition rate of 100%. The experimental results showed that the electronic tongue combined with multivariate statistical analysis can be used to evaluate taste of TCM, at the same time, it could provide a fast and simple method for identifying different processed products.

Efficacy and Material Basis of Shuangjin Sanjie Granules in Treatment on Hyperplasia of Mammary Gland / 中国实验方剂学杂志

Objective:The SD rat model of hyperplasia of mammary gland（HMG） and the ultrahigh-performance liquid chromatography and mass spectrometry (UHPLC-LTQ-Orbitrap MS) technology were used to explore the pharmacological material basis of Shuangjin Sanjie granules (SJSJG) for the treatment on HMG.Method:SD rat models of HMG were administered in groups, and the nipple height and the diameter were measured; the levels of estradiol (E2), progesterone (P) and prolactin (PRL) in serum were detected, pathological examination was conducted for the hyperplasia of breast tissue. Histochemical methods were used to detect the expressions of estrogen receptor α (ERα), androgen receptor (AR), progesterone receptor (PR), tumor necrosis factor-α (TNF-α) proteins. Finally, UHPLC-LTQ-Orbitrap MS technology was used to detect the main chemical constituents of SJSJG, and the pharmacodynamic substance basis was analyzed based on the pharmacological effect.Result:The results of animal experiments showed that compared with the normal group, nipple height and diameter of the model group increased remarkably (P<0.01), serum E2 significantly increased (P<0.01). Pathological examination showed abnormal hyperplasia of breast tissue， expressions of ERα, AR, PR and TNF-α increased， compared with the model group, the nipple height and diameter of the SJSJG group decreased remarkably (P<0.01), serum E2 was decreased significantly (P<0.01)， pathological examination showed weakened abnormal hyperplasia of breast tissue, ERα, AR, PR and TNF-α protein expressions were significant decreased (P<0.01). The results of basic material study showed that 85 chemical components were identified from SJSJG, including 16 alkaloids, 7 flavonoids, 15 terpenes, 9 phenolic acid compounds, 3 coumarin compounds, 10 esters and lactone compounds, 7 fatty acids compounds, 4 amino acids compounds, and 14 other types of ingredients， among them, alkaloids and terpenoids chemical drug substances were closely related.Conclusion:SJSJG can effectively improve the condition of breast hyperplasia, and its medicinal substance basis may include saikosaponin A, Saikosaponin D, verticinone, peimine.

Pharmacological Analysis of Xuanfei Huazhuo Prescription in Treatment of Coronavirus Disease-2019 / 中国实验方剂学杂志

Objective:The targets and signaling pathways of Xuanfei Huazhuo prescription (XFHZP) for the treatment of coronavirus disease-2019 (COVID-19) were explored, and its possible action mechanisms were described through network pharmacology and basic analysis of modern pharmacology. Method:The compounds and targets in XFHZP were collected through TCMSP and BATMAN-TCM databases. The targets of COVID-19 were studied by GeneCards, NCBI and CTD databases. The PPI network was constructed through STRING database. The networks of "herb-meridian" and "traditional Chinese medicines-compounds-targets-disease" were generated by Cytoscape 3.7.0. Then, Kyoto Encyclopedia of Genes and Genomes（KEGG） analysis and Gene Ontology（GO） analysis were made for shared targets through the Omicshare platform. In addition, the disease targets of multiple organ injury, immune injury and severe acute respiratory syndrome (SARS) were retrieved and then mapped with XFHZP. The ratio of intersection targets to XFHZP's targets was calculated. Result:XFHZP has 10 traditional Chinese medicines in total, including 6 medicines with the meridian tropism to lung, 5 medicines with the meridian tropism to the spleen and 5 medicines with the meridian tropism to the stomach. There were 409 compounds and 2 271 targets. There were 8 same inflammatory factors in targets between XFHZP and COVID-19, and each inflammatory factor corresponded to multiple compounds. XFHZP and COVID-19 had 135 intersection targets, and 36 key targets were screened out. A total of 172 signaling pathways were screened out through KEGG signal pathway enrichment (P<0.05). There were 4 000 biological processes, 254 cell components, and 408 molecular functions (P<0.05) according to GO analysis. XFHZP had many common targets with various organ damage targets and immune damage targets, with the ratio of about 7.6%-97.8%. XFHZP had 173 intersection targets with SARS. Conclusion:XFHZP may treat COVID-19 through anti-inflammatory, organ protecting and immune effects. It will provide a certain theoretical basis for the development of drugs for COVID-19.

Clinical Efficacy of Xuanfei Huazhuo Prescription on 40 Cases of COVID-19 / 中国实验方剂学杂志

Objective:To evaluate the clinical efficacy of Xuanfei Huazhuo prescription in the treatment of coronavirus disease-2019 (COVID-19). Method:A total of 40 patients with COVID-19 were selected and treated with Xuanfei Huazhuo prescription. The changes of body temperature, clinical symptoms, computed tomography (CT), blood routine and biochemical indexes were observed before and after treatment. Result:The 40 patients included 15 males and 25 females, with a male to female ratio of 1∶1.7. They were aged between 20-94 years old, with the average age of （43.9±16.3） years old. The course of disease was 8-23 days, with the average of （14±4.4） days. Compared with before administration, the patients' clinical symptoms, such as cough, fever, sputum, diarrhea, loss of appetite and fatigue, were all improved (P<0.05). Before treatment the traditional Chinese medicine (TCM) syndromes of patients were mainly cold dampness lung (57.5%) and cold dampness Lung (42.5%), and the tongue coating was mainly white greasy coating (52.9%). After adjuvant treatment with Xuanfei Huazhuo prescription, the fever removal time was (2.48±2.56) days; white blood cell (WBC), lymphocyte percentage (LYM%), neutrophil percentage (NEUT%), absolute value of lymphocytes (LYM #) indexes of C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), total bilirubin (TBIL), ratio of glutamic oxaloacetic transaminase to glutamic pyruvic transaminase (AST/ALT) and lactate dehydrogenase (LDH) were basically restored to the normal range (P<0.05) compared with before administration. After adjuvant treatment with Xuanfei Huazhuo prescription, the results of three pharyngeal test virus nucleic acid tests were negative, and the lung CT showed that infected lesions were absorbed and all met the discharge criteria. All 40 patients met the discharge criteria and were all cured and discharged, with a cure rate of 100%. There has been no case of recurrence with a positive result of nucleic acid detection so far. The score of symptom and clinical index of patients after administration was （1.62±1.90）, which was significantly lower than that before administration (7.65±4.08, P<0.05). Conclusion:In the adjuvant treatment of COVID-19, Xuanfei Huazhuo prescription can reduce body temperature, promote the absorption of pulmonary inflammation, and improve clinical symptoms, such as fever and cough.

Analysis of "Gansu Prescriptions" in Prevention and Treatment of Coronavirus Disease-2019 / 中国实验方剂学杂志

Coronavirus disease-2019 (COVID-19) is an acute infectious disease caused by a 2019 novel coronavirus (2019-nCoV) infection. It is highly contagious, and can spread quickly home and abroad. It has caused a global pandemic. After the outbreak, Gansu province actively responded to the national "integrated Chinese and western medicine(ICWM)" epidemic prevention policy by organizing an expert group on the prevention and treatment of traditional Chinese medicine(TCM) and establishing a joint working mechanism of ICWM. In adherence to the principle of ICWM, it highlighted the advantages of TCM in epidemic prevention, and emphasized early, timely and whole course use of TCM. The expert group continued to summarize in practice and form a series of "Gansu prescriptions", so as to explore the prevention and control strategy of "prevention in advance, timely interruption and reversal, early prevention and cure, and cure in early stage". Before illness, the prevention shall be made in advance by taking Fuzheng Biwen prescription based on constitution differentiation, in order to strengthen the body resistance and removing pathogenic Qi, after the onset, the syndromes were first treated, interrupted and reversed, and Xuanfei Huazhuo prescription and Qingfei Tongluo prescription were administered based on syndrome differentiation, so as to exorcise pathogenic Qi and cure COVID-19 at the early stage, at the beginning stage of recovery, Jianpi Yifei prescription was used to strengthen the spleen and lungs, and harmonize the stomach and resolve dampness, so as to prevent recurrence. In the principle of ICWM, "Gansu prescriptions" were selected based on the constitution differentiation and syndrome differentiation, so as to prevent the occurrence of epidemics, block light and common symptoms from developing to heavy and critical symptoms, improve the clinical efficacy, shorten the course of disease, and reduce the incidence of critical illness, thereby reducing mortality.

Effect of M1/M2 Alveolar Macrophage Subsets in COVID-19 and Regulatory Mechanism of Traditional Chinese Medicine / 中国实验方剂学杂志

The pathological anatomical results of coronavirus disease-2019 (COVID-19) patients showed that excessive inflammatory reaction in the lungs is one of the important causes for such complications as acute lung injury or acute respiratory distress syndrome. Therefore, regulation of immune response may be an effective measure for COVID-19. Alveolar macrophages have a high heterogeneity and plasticity. The dynamic changes of subsets balance and function of M1/M2 alveolar macrophages have a significant effect on pulmonary inflammatory response during the early and late stages of infection. This paper reviews the classification and function of macrophages and explores the mechanism of alveolar macrophage in the pathological process of COVID-19 at different stages and the pharmacodynamic mechanism of traditional Chinese medicine. Besides, it provides ideas for the treatment of COVID-19 with traditional Chinese medicine and other drugs' research and development based on the regulation of macrophage polarization.

Researches about Transcranial Magnetic Stimulation in Recent Five Years: A Visualization Review Using CiteSpace / 中国康复理论与实践

Results and Conclusion A total of 4820 articles were included. Daskalakis Z J was the author who published the most researches, while it was University of Toronto, United States in term of institutes and nations. The disciplines mostly involved in neuroscience and neurology. The basic researches mostly involved in the transcranial magnetic stimulation and brain motor cortex. The clinical researches mostly involved in stroke and depression. Objective:To analyze the research status and hotspots of transcranial magnetic stimulation in the past five years. Methods:The literatures about transcranial magnetic stimulation in SCI-E of Web of Science from 2014 to 2018 were researched, and the data were analysed with CiteSpace in term of researchers, nations, institutions, disciplines and keywords.

Pharmacodynamics and Mechanism of Chaijin Sanjie Prescription on Rats with Mammary Gland Hyperplasia / 中国实验方剂学杂志

Objective:To investigate the pharmacodynamics and mechanism of Chaijin Sanjie prescription (CJSJP) on rat mammary gland hyperplasia, in order to provide experimental basis for the research and development of new Chinese medicine. Method:SD rat model of mammary gland hyperplasia was established through exogenous intramuscular injection with estrogen and progesterone. After successful establishment of the model, the rats were randomly divided into normal group, model group, and low, medium and high-dose CJSJP groups (3.13, 6.26, 12.52 g·kg-1) and Rupixiao (0.517 g·kg-1) group, with 9 rats in each group. After 28 days of administration, estradiol (E2), progesterone (P) and rolactin (PRL) were measured by radioimmunoassay, uterus and ovary coefficients were calculated; nipple diameter and breast histopathology were observed, estrogen receptor-α(ER-α) expression in mammary gland was measured by immunohistochemistry, and gonadotropin-releasing hormone (GnRH) and gonadotropin-releasing hormone receptor (GnRH-R) mRNA expressions in hypothalamus, pituitary were measured by Real-time PCR. ICR mice were randomly divided into normal group, low, medium and high-dose CJSJP groups (5.2,10.4,20.8 g·kg-1) and Luotongding group (0.038 6 g·kg-1) according to their body weight. Twelve mice in each group were given drugs for 7 days, and 0.6% acetic acid was injected intraperitoneally for 30 minutes after the last administration. The writhing times were observed within 15 minutes. Result:Compared with the normal group, the diameter of nipple was widened, serum E2 was significantly increased (Pα expression were increased in model group. compared with model group, the diameter of nipple was significantly decreased in high dose group of CJSJP (P2 was decreased in all dose groups of CJSJP, pathological score of breast hyperplasia was decreased in middle and high dose groups of CJSJP, GnRH mRNA in hypothalamus was decreased in all dose groups of CJSJP. The writhing times of mice in high dose group of CJSJP was decreased (PConclusion:Chaijin Sanjie prescription can improve the lesions of breast hyperplasia. The therapeutic mechanism may be related to the regulation of GnRH gene expression in hypothalamus and the decrease of estrogen receptor expression.

Clinical features of cow's milk protein allergy in infants presenting mainly with gastrointestinal symptoms: an analysis of 280 cases / 中国当代儿科杂志

OBJECTIVE@#To investigate the epidemiological and clinical features of cow's milk protein allergy (CMPA) in infants presenting mainly with gastrointestinal symptoms.@*METHODS@#A retrospective analysis was performed for the clinical data of 280 hospitalized infants, who were diagnosed with CMPA presenting mainly with gastrointestinal symptoms.@*RESULTS@#Among the 280 infants, 203 infants(72.5%) were aged of less than 6 months. Major manifestations included diarrhea in 171 infants (61.1%), hematochezia in 149 infants (53.2%), vomiting in 71 infants (25.4%), eczema in 57 infants (20.4%), malnutrition in 42 infants (15%) and constipation in 13 infants (4.6%). Of the 280 infants, 258 (92.1%) had mild-to-moderate CMPA and 22 (7.9%) had severe CMPA. Compared with the mild-to-moderate CMPA group, the severe CMPA group had a significantly higher incidence rate of malnutrition (50.0% vs 12.0%) and a significantly lower incidence rate of hematochezia (22.7% vs 55.8%). The breastfeeding CMPA group had significantly lower incidence rates of malnutrition (10.3% vs 24.6%) and severe CMPA (4.4% vs 18.0%) than the artificial feeding CMPA group, and the artificial feeding CMPA group had a significantly lower incidence rate of hematochezia than the breastfeeding and mixed feeding CMPA groups (37.7% vs 56.6%/59.0%).@*CONCLUSIONS@#CMPA presenting mainly with gastrointestinal symptoms is more common in infants aged of less than 6 months. Diarrhea and hematochezia are the most common manifestations at the time of onset. Most infants have mild-to-moderate allergy. Compared with breastfeeding, artificial feeding is more likely to cause malnutrition and severe CMPA.

Prokaryotic expression and purification of recombinant CARD9-MBP fusion protein / 医学研究生学报

Objective　CARD9 can activate several pathways involved in immunity, such as NF-ΚB, MAPK, etc. However the mechanism of this process has not yet been elucidated. For conducting relevant experiments in vitro, a prokaryotic expression vector of CARD9-MBP fusion protein has to been construct, and the fusion protein need to be expressed and purified.　Methods　The coding sequence of CARD9 and MBP genes were amplified by PCR and the recombinant plasmid was correctly inserted into the pET-30a(+) vector. The recombinant plasmid was transformed into E.coli DH5α competent cells and proceeded PCR identification, restriction analysis and gene sequencing. The correct recombinant plasmid was transformed into E.coli BL21(DE3) competent cells. The target protein was induced to express by IPTG under different conditions. Relative molecular weight of the target protein was detected by SDS-PAGE electrophoresis. The CARD9-MBP fusion protein was purified by MBP maltose chromatography column and gel filtration chromatography column, and identificated by MALDI-TOF mass spectrometry after MBP-tag to be removed by HRV3C enzyme.　Results　The CARD9-MBP fusion protein was successfully constructed and confirmed by PCR and restriction analysis. The result of gene sequencing was consistent with the target sequence. The SDS-PAGE electrophoresis showed that the target protein with molecular mass (MR) about 105 000 was successfully induced to express in E.coli BL21 (DE3). A quite pure CARD9-MBP fusion protein was obtained by purification of MBP maltose chromatography column. Identification by MALDI-TOF mass spectrometry demonstrated that the target protein after MBP-tag to be removed by HRV3C enzyme is CARD9 protein. In the later stage, gel filtration chromatography column was used to obtain further pure CARD9-MBP fusion protein.　Conclusion　The prokaryotic expression vector of CARD9-MBP fusion protein was successfully constructed and a large number of soluble protein expressed. The purified target protein can be obtained by purification with MBP maltose chromatography column and gel filtration chromatography column.

Effect of Acinetobacter baumannii biofilm on antimicrobial resistance / 中国感染控制杂志

Objective To understand biofilm-forming ability of Acinetobacter baumannii(A.baumannii) in clinical isolates,and evaluate the effect of biofilm formation on antimicrobial resistance.Methods A total of 47 A.baumannii clinical isolates were collected,biofilm-forming ability was detected by crystal violet staining assay,the minimal inhibitory concentration(MIC) of 12 kinds of antimicrobial agents to biofilrn forming isolates were detected when the isolates were in planktonic or biofilm formatted condition;biofilm-forming ability of positive isolates at different culture condition (shaking or stable) and on different materials(glass tube or polypropylene plastic centrifugal tube) were detected.Results Thirteen isolates (27.66％) exhibited biofilm formation.In planktonic condition,antimicrobial resistance rates of these isolates were all ＜35％,no isolates shown resistance to polymyxin and tigecycline,in biofilm formatted condition,resistance rates to tigecycline was 15.38％,resistance rates to the other antimicrobial agents were all 100％.Of 13 tested isolates,≥10 isolates shown higher absorbance in the shaking condition than in the stable condition,≥10 tested isolates shown higher absorbance in polypropylene plastic centrifugal tube than in the glass tube.Conclusion The resistance of A.baumannii to antimicrobial agents is greatly changed after the formation of biofilm,which may results in the failure of antimicrobial therapy for infection.

Promoting cell viability and migration of gastric cancer cells by PRRX2 via activation of Wnt/β-catenin signaling pathway / 中国病理生理杂志

AIM:To study the effect of paired-related homeobox 2(PRRX2)gene on the viability and migra-tion ability of gastric cancer cells,and to analyze the underlying mechanism of regulating Wnt /β-catenin signaling pathway. METHODS:The expression of PRRX2 in gastric cancer and normal gastric tissue and the correlation between PRRX 2 ex-pression in gastric cancer tissues with the overall survival rate of gastric cancer patients were analyzed by bioinformatics. The small interfering RNA(siRNA)and over-expressed plasmids of PRRX2 were transfected into gastric cancer cells MGC-803 and SGC-7901,respectively.MTT assay and Transwell assay were used to detect the viability and migration ability of gastric cancer cells.Western blot and TOPflash/FOPflash dual-luciferase reporter gene assay were used to detect the activi-ty of Wnt/β-catenin signaling pathway.Co-immunoprecipitation was used to detected the interaction between PRRX 2 and β-catenin proteins.RESULTS:Knockdown of PRRX2 attenuated the viability and migration ability of gastric cancer cell line MGC-803(P <0.05).Over-expression of PRRX2 enhanced the viability and migration ability of SGC-7901 cells (P<0.05),increased the protein levels of β-catenin, c-Myc and cyclin D1(P<0.05)and the activity of TOPflash/FOPflash dual-luciferase reporter gene(P<0.05).PRRX2 interacted with β-catenin protein in gastric cancer cells.CON-CLUSION:PRRX2 promotes the viability and migration ability of gastric cancer cells, which may be related to Wnt/β-catenin signaling pathway.

Histological changes of cornea in vivo of patients with xerophthalmia / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To investigate the changes of corneal cytology of patients with different degrees of xerophthalmia of the types of deficient aqueous production and over evaporation with confocal microscopy. METHODS:Retrospective case series was adopted with three - dimensional corneal topography and anterior segment analysis system (Sirius system). A total of 33 typical dry eye patients (total 65 eyes) with deficient aqueous production and 28 typical dry eye patients (total 55 eyes) with over evaporation were selected from optical center of the First Affiliated Hospital of Harbin Medical University from December 2016 to June 2017, which were grading according to dry eye degrees. The corneal epithelial cell (surface epithelial cell, pterygoid epithelial cell, basal epithelial cell), Langerhans cell (dendritic cell),stroma (deep stromal layer and superficial stromal layer), the density and form of endothelial cells were observed and analyzed with the confocal microscope. RESULTS: Confocal microscope showed that the number of corneal epithelial cells decreased with the increasing of dry eye severity in patients with tear deficiency and evaporative. The results were statistically significant (P<0.01). The sizes of each layer of corneal epithelium cells were uneven, and the arrangement was irregular. The diameter of corneal sub-epithelial nerve fiber of deficient aqueous production dry eye became thinner with irregular arrangement. With the increase in severity of dry eye,the degree of curvature and branches gradually increased. The activation degree of Langerhans cells of evaporative dry eye was directly proportional to the severity of dry eye(P<0 01). There was no significant changes in the number of superficial stromal cells in the two types of dry eye (P>0 05). The number of deep stromal cells in the two types of dry eye had no relations with the severity of dry eye (P>0. 05). There was no significant changes in the number of corneal endothelial cells in the two types of dry eye (P>0.05). CONCLUSION:The Sirius system can be used for non-contact and non-invasive examination and classification. And there is changes of the density and morphology in each layers of cornea cells of two types of dry eye with the confocal microscope.

Research progress of exosomes in treating spinal cord injury / 基础医学与临床

Exosomes are membrane vesicles secreted by cells into the extracellular environment. Exosomes are a new cell component in the treatment of spinal cord injury. Exosomes are secreted by mesenchymal stem cells have anti-apoptotic, anti-inflammatory and pro-angiogenic effects in the treatment of spinal cord injury, which play a key role in nerve cell-cell communication and can delivery exogenous genetic material. Exosomes can penetrate blood-brain barrier and are more stable than their parent cells, reducing the safety risks in the administration of viable cells.

Effect of lipoxin A4 on the expression of Toll-like receptor 4 and TNF receptor-associated factor 6 in the liver of obese rats with sepsis / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the protective effect of lipoxin A4 (LXA4) against sepsis induced by lipopolysaccharide (LPS) in rats with obesity and its effect on the expression of Toll-like receptor 4 (TLR4) and TNF receptor-associated factor 6 (TRAF6) in the liver.</p><p><b>METHODS</b>A total of 60 male Sprague-Dawley rats aged three weeks were randomly divided into a normal group and an obesity group, with 30 rats in each group. A rat model of obesity was established by high-fat diet. Each of the two groups was further randomly divided into control group, sepsis group, and LXA4 group, and 8 rats were selected from each group. The rats in the control, sepsis, and LXA4 groups were treated with intraperitoneal injection of normal saline, LPS, and LXA4+LPS respectively. Twelve hours later, blood samples were collected from the heart and liver tissue samples were also collected. ELISA was used to measure the serum levels of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α). Western blot was used to measure the protein expression of TLR4 and TRAF6 in liver tissue. Quantitative real-time PCR was used to measure the mRNA expression of TLR4 and TRAF6.</p><p><b>RESULTS</b>After being fed with high-fat diet for 6 weeks, the obesity group had significantly higher average weight and Lee's index than the normal group (P<0.05). Compared with the normal group, the obesity group had significant increases in the serum levels of IL-6 and TNF-α (P<0.05). In the normal group or the obesity group, the sepsis subgroup had significant increases in the serum levels of IL-6 and TNF-α compared with the control subgroup (P<0.05), while the LXA4 subgroup had significant reductions in the two indices compared with the sepsis subgroup (P<0.05). Compared with the normal group, the obesity group had significant increases in the protein and mRNA expression of TLR4 and TRAF6 (P<0.05). In the normal group or the obesity group, the sepsis subgroup had significant increases in the protein and mRNA expression of TLR4 and TRAF6 compared with the control subgroup (P<0.05). Compared with the sepsis subgroup, the LXA4 subgroup had significant reductions in the protein and mRNA expression of TLR4 and TRAF6 (P<0.05).</p><p><b>CONCLUSIONS</b>LXA4 can reduce the serum levels of IL-6 and TNF-α and alleviate inflammatory response. LXA4 can inhibit the expression of TLR4 and TRAF6 in the liver of septic rats, possibly by inhibiting the TLR4 signaling pathway.</p>