Genotypic and phenotypic analysis in a family affected with sector retinitis pigmentosa and macular dystrophy caused by Spermatogenesis-associated protein 7 gene / 中华眼底病杂志

Objective:To analyze the pathogenic gene and clinical phenotypes of a family affected with rare sector retinitis pigmentosa (sector RP).Methods:A retrospective clinical study. A patient with sector RP diagnosed in Renmin Hospital of Wuhan University and his parents were included in the study. Detailed medical history was collected; best corrected visual acuity (BCVA), fundus color photography, autofluorescence (AF), visual field, optical coherence tomography (OCT), electroretinogram, fluorescein fundus angiography (FFA), indocyanine green angiography (ICGA) examination were performed. The peripheral venous blood of the patient and his parents were collected, and DNA was extracted. A whole exon sequencing was used for the proband. The mutations were verified by targeted Sanger sequencing and quantitative polymerase chain reaction. Bioinformatics analysis and cosegregation analysis were performed.Results:The proband, a 17-year-old male, had presented with gradually decreased vision in the past 2 years with BCVA of 0.4 in both eyes. Retinal vessels attenuation and macular dystrophy without obvious pigmentation on the fundus were observed. AF showed, in bilateral eyes, a symmetrical hypo-autofluorescent region only in the inferonasal quadrant and "petal-like" hyper-AF macula. The visual field examination showed defects in the superotemporal quadrant corresponding to the affected retina. OCT showed loss of the photoreceptor layer except for the foveal region. Electroretinogram examination presented reduced scotopic wave peaks and extinct photopic response. FFA and ICGA showed the atrophy retinal pigment epithelium around the optic disk and in the inferior retina. The clinical phenotypes of the parents were normal. The whole exon sequencing identified one mutation in SPATA7 gene, c.1112T>C (p.Ile371Thr) in exon10 and a copy number variation in trans. The missense mutation resulted in the change of isoleucine to threonine at amino acid 371 in the encoded SPATA7 protein, and the mother carried this heterozygous mutation c.1112T>C. According to the guidelines of the American College of Medical Genetics and Genomics (ACMG) criteria and guidelines for classification of genetic variants, the missense mutation was classified as the uncertain significance. The CNV, originating from his father, contributed to the loss of exon10 and was confirmed as the likely pathogenic variant. Conclusions:The macula can be involved in sector RP, leading to the macular dystrophy. The missense variant in SPATA7 gene, c.1112T>C (p.Ile371Thr), might be a pathogenic mutation site in this pedigree.

Survival and disease burden trend analysis of occupational pneumoconiosis from 1963 to 2020 in Shizuishan City / 中华劳动卫生职业病杂志

Objective: To understand the survival status and its influencing factors of occupational pneumoconiosis patients in Shizuishan City, and to analyze the disease burden of occupational pneumoconiosis and its trend, so as to provide scientific basis for formulating comprehensive prevention and treatment measures of occupational pneumoconiosis. Methods: A retrospective survey was conducted during July to December 2020 to explore the survival status of occupational pneumoconiosis patients who had been reported from 1963 to 2020 in Shizuishan City. The Kaplan-Meier method and Life-table method were used for survival analysis, and Cox proportional hazards regression model was used to analyze the influencing factors of survival time. The disability adjusted life years (DALY) was applied to analyze the disease burden of occupational pneumoconiosis and its temporal trend. Results: From 1963 to 2020, a total of 3263 cases of occupational pneumoconiosis were reported in Shizuishan City, of which 1467 died, so that the fatality rate was 44.96%. The median survival time was 26.71 years, average age of death was (70.55±10.92) years old. There were significant differences in the survival rates of occupational pneumoconiosis patients among different types, diagnosis age, exposure time, industry, initial diagnosis stage and whether upgraded (P<0.05) . As the survival time increased, the survival rate of patients decreased gradually. When the survival time was ≥50 years, the cumulative survival rate of patients was 4.20%. Cox regression analysis suggested that the type of pneumoconiosis, industry, diagnosis age, exposure time, initial diagnosis stage and whether upgraded were the influencing factors for the survival time of patients with occupational pneumoconiosis (P<0.05) . The total DALY attributable to occupational pneumoconiosis from 1963 to 2020 in Shizuishan City was 48026.65 person years, of which the years of life lost (YLL) was 15155.39 person years, and the average YLL was 10.33 years/person, and the years lost due to disability (YLD) was 32871.26 person years, and the average YLD was 10.07 years/person. The DALY attributed to coal worker's pneumoconiosis and silicosis were 39408.51 person years and 6565.02 person years, respectively, and they accounted for 82.06% and 13.67% of the total disease burden in Shizuishan City, respectively. The DALY caused by occupational pneumoconiosis in the age group of 40-49 years old and the first diagnosis of stage I occupational pneumoconiosis were higher, which were 20899.71 and 36231.97 person years, respectively. The average YLL and average YLD showed a volatility downtrend over time. Conclusion: The disease burden of occupational pneumoconiosis cannot be ignored in Shizuishan City, and timely targeted measures should be taken for key populations and key industries. It is recommended that life-cycle health management and hierarchical medical should be taken to improve the life quality of patients and prolong their lifes.

Optimization of Extraction and Purification Process of Total Iridoids and Total Crocins in Gardeniae Fructus / 中国实验方剂学杂志

ObjectiveTo optimize the extraction and purification process of Gardeniae Fructus for industrial production， and to obtain the total iridoid and total crocin extracts. MethodOrthogonal test was used to optimize the water extraction process by taking contents of geniposide， genipin gentiobioside， gardenoside， crocin-1 and crocin-2 as indicators and the decocting time， decocting times and water amount as factors. The purification process was optimized by single factor test， and four different types of macroporous adsorption resins were screened. The process conditions such as resin type， maximum loading amount， water washing amount， ethanol concentration， ethanol dosage， and flow rate of sample loading were mainly investigated. In addition， the drying methods （vacuum drying and spray drying） of the extract were investigated， and a pilot scale-up verification test was carried out. ResultThe optimal water extraction process of Gardeniae Fructus was to add 15， 10 times the amount of water for decocting twice， 1 h each time. The optimal purification process was as follows：the water extract through SP825L macroporous resin column， the amount of crude drug-the amount of resin （1∶1.5）， the sample loading flow rate of 3 BV h-1， adding 2 BV of water to remove impurities， adding 4 BV of 30% ethanol to obtain the iridoid part， then adding 3 BV of 70% ethanol to obtain the crocin part， collecting the ethanol lotion， and drying at 70 ℃. Under these conditions， the extraction amount of total iridoids was 590.75 mg·g-1 with the transfer rate of 70.48%， and the yield of dry extract was 8.89%. The extraction amount of total crocins was 83.37 mg·g-1 with the transfer rate of 22.20%， and the dry extract yield was 2.60%. ConclusionThe optimized extraction and purification process is stable and feasible with high extraction rate of active components， which is suitable for the industrial extraction and purification of active parts of Gardeniae Fructus.

Efficacy and Mechanism of Yishen Shengjing Prescription in Treatment of Oligoasthenospermia / 中国实验方剂学杂志

ObjectiveWe aimed to investigate the efficacy and mechanism of Yishen Shengjing Prescription （YSP） in the treatment of oligoasthenospermia in rats. MethodThe oligoasthenospermia rat model was established by injection with cyclophosphamide （35 mg·kg-1·d-1） for 5 consecutive days. Rats were randomly assigned into control group （without treating with cyclophosphamide）， model group， low- （YSP-L）， medium- （YSP-M）， and high- （YSP-H） dose （2.91， 5.83， and 11.66 g·kg-1， respectively） groups， Wuzi Yanzongwan （WYW， 1.03 g·kg-1） group， and L-carnitine （0.17 g·kg-1） group， with 8 rats in each group. After 28 days of drug intervention， the body weight， testicular weight， and testicular index of rats were recorded. The sperm quality in epididymis was detected by computer-assisted sperm analysis （CASA） system. Hematoxylin-eosin （HE） staining was employed for observation of testicular tissue morphology. The levels of malondialdehyde （MDA） and superoxide dismutase （SOD） in testicular tissue were detected by colorimetry. The levels of follicle-stimulating hormone （FSH）， luteinizing hormone （LH）， and testosterone （T） in serum were determined by enzyme-linked immunosorbent assay（ELISA）. TdT-mediated dUTP nick-end labeling （TUNEL） was employed to detect the apoptosis of testicular cells. The protein levels of B cell lymphoma-2（Bcl-2）， Bax， and cleaved Caspase-3 in testicular tissue were detected by Western blot. ResultCompared with the control group， the model group showed decreased body weight， testicular weight and index， sperm concentration and motility （P<0.01） and increased testicular pathological score （P<0.01）. Compared with the model group， the YSP-M， YSP-H， WYW， and L-carnitine groups showed increased body weight， testicular weight， testicular index， sperm concentration and motility and decreased testicular pathological score. After modeling， the SOD level decreased （P<0.01） while the MDA content increased （P<0.01） in the testicular tissue. YSP-H， WYW， and L-carnitine reversed the SOD and MDA level changes caused by modeling. Compared with the control group， the model group exhibited declined T level （P<0.01） and increased FSH and LH levels （P<0.01）. Compared with the model group， YSP， WYW， and L-carnitine increased the T level （P<0.01） and decreased the LH level （P<0.05， P<0.01）. The apoptosis rate of spermatogenic cells in the model group was higher than that in the control group （P<0.01）， whereas YSP-M， WYW， and L-carnitine reversed such changes （P<0.01）. The model group rats showed decreased expression of Bcl-2（P<0.05） and increased expression of Bax and cleaved Caspase-3 （P<0.05， P<0.01）. Compared the model group， YSP-M， YSP-H， WYW， and L-carnitine up-regulated the Bcl-2 expression and down-regulated the cleaved Caspase-3 expression （P<0.05， P<0.01）. ConclusionYSP improved the sperm quality of oligoasthenospermia model rats by regulating the antioxidant system and sex hormone levels and inhibiting the apoptosis of spermatogenic cells.

Views on Key Technical Problems in Research and Development of Famous Classical Formulas / 中国实验方剂学杂志

In this paper， the key technical problems in the research and development of famous classical formulas are analyzed. Firstly， the puzzled problem for a long-time， which is conversion relationship from medicinal metrology of Han dynasty （HD） to that of modern （gram，g）， is comprehensively expounded that one Liang （两） of HD=3 g is more appropriate. Secondly， the model and principles of quality consistency evaluation are given for the transformation from the quality of authoritative basic sample prepared by casserole （ABS-C） to the quality consistency in Laboratory process， pilot-scale process and industrial production. The consistency evaluation model is ξABS-X=K1（Q1ABS-X/Q1ABS-C）+K2（Q2ABS-X/Q2ABS-C）+……+Ki（QiABS-X/QiABS-C）=∑Ki（QiABS-X/QiABS-C）（i=1，2，3……n）. In the formula， ABS-X means laboratory reference sample ABS-C （ABS-L）， pilot-scale ABS-C （ABS-mP） or industrial production ABS-C （ABS-P）， ξABS-X means the quality consistency rate or similarity degree of ABS-L， ABS-mP and ABS-P processes with ABS-C， Ki means the weight of each quality evaluation index （i）， QiABS-X is the data of i in ABS-L， ABS-mP， ABS-P samples， and QiABS-C is the data （or mean） of i in ABS-C sample. Thirdly， in order to control the quality of the herbal medicines whose active ingredients were unknown， their chemical constituents should be studied deeply， and if necessary， the bioassay research should be carried out according to the main efficacy or indication of famous classical formulas. Finally， for the special processing of some herbal medicines， it is difficult to formulate the processing method， technology and standard of prepared slices. It is suggested that the scientific connotation and historical evolution of the special processing method should be thoroughly sorted out， and its technological characteristics are summarized， the modern processing technology and production processes are simulated， and then the corresponding processing methods and quality standards are formulated.

Anti-inflammatory Effects of Yuyang Capsule on Bacterial Dermatitis Model Rats and Its Effect on TLR/NF-κB Signaling Pathway / 中国药房

OBJECTIVE：To study the anti-inflammatory effects of Yuyang capsule on bacterial dermatitis model rats and its effect on TLR/NF-κ B pathway. METHODS：Minimum inhibition concentration （MIC）and minimal bactericidal concentration （MBC）of Yuyang capsule against Staphylococcus aureus were determined by microdilution test. Totally 50 SD rats were randomly divided into model group ，positive control group （amoxicillin and clavulanate potassium ，120 mg/kg as amoxicillin ），Yuyang capsules high-dose ，medium-dose and low-dose groups （according to MIC ），with 10 rats in each group. The model of bacterial dermatitis was established by using the burned skin of rats infected with S. aureus . 24 h after modeling ，administration groups were intragastrically given the corresponding drug ，and model group was intragastrically given the same amount of normal saline ，once a day，for consecutive 7 days. The skin healing rate was calculated on the 1st，3rd，5th and 7th day of administration ，and the scab formation，decrustation and healing were recorded. The contents of IL- 1β，IL-6，IL-10，TNF-α，hydroxyproline（HYP），collagen Ⅰ（Col Ⅰ）and Col Ⅲ in skin tissue were detected by ELISA. Morphology changes of skin tissues were observed by HE staining. The ultrastructure was observed by transmission electron microscope. Protein expression of TLR 2，TLR4 and p-NF-κB p65 were detected by Western blotting assay. RESULTS ：MIC and MBC of Yuyang capsule were 25 mg/mL and 50 mg/mL，respectively. Dose of Yuyang capsules high-dose ，medium-dose and low-dose groups were set at 600，300，150 mg/kg. Compared with model group，scab appeared on the injured skin 3 days after administration in Yuyang capsule high-dose and medium-dose groups ，and decrustation appeared on the injured skin of part mice 5-7 days after administration ；the skin healing rate of the positive control group，Yuyang capsule high-dose and medium-dose groups were all significantly increased at each time point. The contents of IL- 1 β，IL-6，IL-10 and TNF-α，pathological score ，protein expression of TLR 2，TLR4 and p-NF-κB p65 were decreased significantly （P＜0.05 or P＜0.01）；the pathological changes such as inflammatory cell infiltration in skin tissue were improved. The contents of HYP，Col Ⅰ and Col Ⅲ were increased significantly in positive control group and Yuyang capsule high-dose group （P＜0.05 or P＜0.01）. There was no statistical significance in most above indexes in positive control group ，Yuyang capsule high-dose and medium-dose groups （P＞0.05）. CONCLUSIONS ：Yuyang capsule can promote skin healing of bacterial dermatitis model rats and shows certain anti-inflammatory effects ；the mechanism may be related to inhibiting TLR/NF-κB signaling pathway and reducing inflammatory response.

Research and development strategies of new Chinese medicines based on new regulations / 中国中药杂志

In recent years, only a small number of new Chinese medicines have been approved for marketing, which has embodied the bottleneck in the development of the Chinese medicine industry. To tackle this problem, the National Medical Products Administration has issued a series of regulations and technical requirements. In the context of new regulations, this study deeply explored the research and development strategies of new Chinese medicines under the guidance of the new classification of drug registration, and discussed the key technical issues in the research and development.

Clinical features and gene analysis in a family with type 2 congenital generalized lipodystrophy due to BSCL2 mutation / 中华内分泌代谢杂志

Objective:To investigate the clinical and genetic features in a family with type 2 congenital generalized lipodystrophy, and to improve the understanging of this disease.Methods:The clinical symptoms, results of the laboratory, and radiography examinations of the patient and his family members were analyzed. The whole exome sequencing and Sanger validation were used to determine the genetic cause of the disease.Results:Generalized lipodystrophy, impaired liver function, severe hypertriglyceridemia, and acanthosis nigricans were found in the proband. His serum leptin level was much lower than normal value. The proband and three members of this family were confirmed to have insertion mutation at exon 5 of BSCL2 gene. The site was mutated from TTC to TCGGTC, resulting in the replacement of glutamate by aspartate and arginine. The mutation in proband was homozygote, and his father, mother, and brother were heterozygous.Conclusions:The mutation in exon 5 c. 545_546insCCG of BSCL2 gene leads to the occurrence of type 2 congenital generalized lipodystrophy.

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.

Research Status of Zexietang / 中国实验方剂学杂志

Zexietang is derived from Jingui Yaolue （《金匮要略》）， which is composed of Alismatis Rhizoma and Atractylodis Macrocephalae Rhizoma， and has the effect of inducing diuresis and invigorating the spleen to produce water. Compared with western medicine in the treatment of related diseases， Zexietang can not only improve the curative effect， but also reduce the occurrence of adverse reactions， so as to achieve long-term stable administration. The authors sorted out and analyzed the chemical composition， pharmacological effect and clinical application of Zexietang in recent years. It was found that the main active components of Zexietang were alismol A and B， 23-acetyl-alismol B and C， atractylenolides （atractylenolide Ⅰ， Ⅱ， Ⅲ） and polysaccharides. Pharmacological experiments showed that they had diuretic， hypolipidemic， anti-inflammatory and others. And it can be used in the treatment of hypertension， hyperlipidemia， vertigo， cerebral vascular insufficiency and other diseases combined with other Chinese materia medica， and the curative effect is obvious. By summarizing the research status of Zexietang in recent years， its active components and pharmacological mechanism can be further clarified， which provides the basis for the clinical application of Zexietang and guides the direction of its further research.

Research Progress of Targeted Drug Delivery Nanocarriers Through Blood-brain Barrier / 中国实验方剂学杂志

The small size， moderate drug loading， and targeting properties of nano-preparations make them can be excellent delivery tools for drugs， genes or proteins crossing the cell or blood-brain barrier （BBB）. Currently， facilitating drug crossing BBB with innovative nano-drug delivery systems is considered as a strategic approach for the prevention， diagnosis and treatment of central nervous system （CNS） diseases. However， with the deepening of the research， the adverse reactions and toxicity of nanocarriers have gradually attracted the attention of researchers. Based on this， this paper summarized the situation of BBB-penetrating targeted nano-preparations at home and abroad in recent years from the perspective of classification of types and properties of nanocarriers， and analyzed the advantages and disadvantages of each carrier. The results showed that nano-preparations with active ingredients of traditional Chinese medicine （TCM） as carriers have become a promising way of cancer treatment， but the complexity and diversity of TCM components limited its application to a certain extent. Further studies should be strengthened to lay a foundation for the application and development of TCM nano-preparations in the field of CNS diseases.

Analysis of "Tianyou (TE 16) and five regions" in the human body / 中国针灸

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Preparation of paclitaxel-loaded and folic acid-modified poly (lactic-co-glycolic acid) nano-micelles and in vitro anticancer effect on cervical cancer HeLa cells / 中国中药杂志

The paclitaxel-loaded and folic acid-modified poly(lactic-co-glycolic acid) nano-micelles(PTX@FA-PLGA-NMs) were prepared by the emulsion solvent evaporation method, and the parameters of paclitaxel-loaded nano-micelles were optimized with the particle size and PDI as evaluation indexes. The morphology of the nano-micelles was observed by transmission electron microscopy(TEM), and the stability, drug loading and encapsulation efficiency were systematically investigated. In vitro experiments were performed to study the cytotoxic effects of nano-micelles, apoptosis, and cellular uptake. Under the optimal parameters, the nano-micelles showed the particle size of(125.3±1.2) nm, the PDI of 0.086±0.026, the zeta potential of(-20.0±3.8) mV, the drug loading of 7.2%±0.75%, and the encapsulation efficiency of 50.7%±1.0%. The nano-micelles were in regular spherical shape as observed by TEM. The blank FA-PLGA-NMs exhibited almost no inhibitory effect on the proliferation and growth of tumor cells, while the drug-loaded nano-micelles and free PTX exhibited significant inhibitory effects. The IC_(50) of PTX@FA-PLGA-NMs and PTX was 0.56 μg·mL~(-1) and 0.66 μg·mL~(-1), respectively. The paclitaxel-loaded nano-micelles were potent in inhibiting cell migration as assessed by the scratch assay. PTX@FA-PLGA-NMs had good pro-apoptotic effect on cervical cancer HeLa cells and significantly promoted the uptake of HeLa cells. The results of in vitro experiments suggested that PTX@FA-PLGA-NMs could target and treat cervical cancer HeLa cells. Therefore, as nanodrug carriers, PTX@FA-PLGA-NMs with anti-cancer activity are a promising nano-system for improving the-rapeutic effects on tumors.

Acute fever with rash, necrosis, and bullae in both lower extremities in a 12-year-old girl / 中国当代儿科杂志

A girl, aged 12 years, was admitted due to fever and rash for 3 days. The child developed recurrent high fever and rash on both lower extremities 3 days before, and the rash on left lower extremity quickly merged into a patch within 24 hours, with hemorrhage and necrosis in black and purple, large vesicles, and blisters in the center. Laboratory examination showed a reduction in platelet count and significant increases in fibrinogen and D-dimer during the course of the disease. The child was diagnosed with purpura flulminans. She was given meropenem combined with linezolid for anti-infection, injection of gamma globulin for immunoregulation, and low-molecular-weight heparin for anticoagulation. The fluid in the rash blisters was drawn and the wound was treated to prevent infection. The child's temperature returned to normal, with improvement in gangrene. She was discharged after platelet count, fibrinogen, and D-dimer had returned to normal. Purpura fulminans is a rare thrombotic hemorrhagic disease with rapid progression and is commonly seen in children. Without timely treatment, it may cause severe sequelae with high disability and mortality rates. Anti-infection, correction of coagulation function, and local management of gangrene skin are of great importance during treatment.

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.

Preparation and Evaluation of β-Cyclodextrin Inclusion Complex of Volatile Oil in Couplet Medicines of Moslae Herba and Pogostemonis Herba / 中国实验方剂学杂志

Objective::To investigate the pharmacodynamics of volatile oil in couplet medicines of Moslae Herba and Pogostemonis Herba, to establish a method for simultaneous determination of three essential ingredients (thymol, carvacrol and patchouli alcohol) in volatile oil of the couplet medicines by gas chromatography (GC), to optimize the preparation process of β-cyclodextrin (β-CD) inclusion complex of volatile oil in the couplet medicines and to confirm the formation of the inclusion complex. Method::An in vitro inflammatory response model was established by hyaluronidase activity inhibition test in order to detect the anti-inflammatory activity of the volatile oil. Also, the antioxidant activity of the volatile oil was assessed by 1, 1-diphenyl-2-trinitrophenylhydrazine (DPPH) free radical scavenging method. The inclusion of volatile oil in couplet medicines of Moslae Herba and Pogostemonis Herba was prepared by scaturated aqueous solution method, colloid milling method and grinding method, respectively. GC was used to determine the contents of thymol, carvacrol and patchouli alcohol in volatile oil for optimizing extraction and inclusion processes of volatile oil. Scanning electron microscope, infrared spectroscopy, thermal differential analysis, and X-ray diffraction (XRD) were used to verify the formation of the inclusion complex. Result::The volatile oil not only inhibited hyaluronidase activity to a certain extent, but also eliminated DPPH and increased with the increase of concentration. There was a good linear relationship between the peak area and concentration of thymol, carvacrol and patchouli alcohol at 0.021 3-0.426, 0.020 04-0.400 8, 0.022 6-0.452 g·L-1 (R2>0.999), respectively. Their recoveries were 99.59%(RSD 1.6%), 100.15%(RSD 1.5%), 100.70%(RSD 1.4%), respectively. The colloid milling method was optimized, and the formation of the inclusion complex was verified by the aforementioned methods. Conclusion::The volatile oil in couplet medicines of Moslae Herba and Pogostemonis Herba has certain anti-inflammatory activity and anti-oxidation ability. The colloid milling method was the best inclusion process for the volatile oil. The established GC has the advantages of simple, sensitive, accurate, reliable and reproducible, which can meet the requirements of simultaneous determination of thymol, carvacrol and patchouli alcohol in the inclusion complex.

Different expression levels of exosomal miR-503 in peritoneal dialysis effluent from patients of different peritoneal transport characteristics and bioinformatics analysis / 中华肾脏病杂志

Objective:To compare the expression level of exosomal miR-503 in peritoneal dialysis effluent (PDE) from patients of different peritoneal transport characteristics, predict the target genes of miR-503 and provide bioinformatic data for researches of peritoneal transport characteristics.Methods:Twenty-four stable peritoneal dialysis (PD) patients were selected and divided into high transport group (H group, n=12) and low transport group (L group, n=12) according to the results of peritoneal equilibration tests (PET). The 500 ml PDE that was left on the patient's abdomen overnight was collected and concentrated using ultrafiltration cell. Exosomes in PDE were resuspended in phosphate buffered saline (PBS) after ultracentrifugation and the characteristics of PDE exosomes were identified by transmission electron microscope (TEM), nanoparticle tracking analysis (NTA), Western blotting and fluorescent staining. MicroRNAs were extracted from PDE exosomes. The expression levels of PDE exosomal miR-503 in the two groups were detected by quantitative real-time PCR. Then the relations between the relative quantity of PDE exosomal miR-503 and PET values or 24 h ultrafiltration volume (UF) were analyzed. Targetscan and miRDB databases were used to predict the target genes of miR-503. Gene ontology (GO) functional enrichment and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway analysis were relied on DAVID (https://david.ncifcrf.gov/). Results:The exosomes in PDE showed a round and cup-shaped morphology under TEM, and the diameters were approximately 100 nm measured by NTA. The specific biomarkers of exosomes, CD63, CD81 and heat shock protein -70 (HSP-70) were all detected by Western blotting. The internalization and uptake of the exosomes was observed after fluorescent staining. The relative expression level of PDE exosomal miR-503 in H group was found to be significantly higher than that in L group ( P=0.002), and the relative quantity of PDE exosomal miR-503 was significantly positively correlated with PET values ( r=0.547, P=0.006), but not 24 h UF ( r=-0.297, P=0.159). There were 156 target genes of miR-503 in total that could be predicted by two different databases at the same time. GO analysis of these 156 target genes was mainly focused on kinase binding, regulation of protein modification and catabolic process as well as regulation of epithelial cell proliferation. KEGG enriched many tumor associated or classical signaling pathways, including transforming growth factor-β (TGF-β) signaling pathway and vascular endothelial growth factor (VEGF) signaling pathway. The prediction showed that vascular endothelial growth factor A (VEGFA) was a direct target gene of miR-503 and it was also related to many proteins involved in fibrosis mechanism. Conclusions:The expression level of PDE exosomal miR-503 is significantly higher in H group, and positively correlates with PET values, which may regulate the angiogenesis of peritoneal vessels by targeting VEGFA.

Changes of brain function in resting-state in patients with nonproliferative diabetic retinopathy assessed by fractional low-frequency amplitude technique / 中华实验眼科杂志

Objective:To study the changes of brain function in patients with diabetic retinopathy (DR) by using resting-state functional magnetic resonance imaging (rs-fMRI) based on the fractional amplitude of low-frequency fluctuation (fALFF) method.Methods:A cross-sectional study method was performed, 34 cases of non-proliferative DR patients treated in the Renmin Hospital of Wuhan University during July 2018 to March 2019 were collected.Meanwhile, 34 healthy volunteers with matched age, gender, and education years were recruited.All subjects were performed with conventional MRI scanning and rs-fMRI scanning, then the fALFF values in the whole brain were calculated, and the differences of neuronal activities between the two groups were explored.This research protocol was approved by the Ethic Committee of Renmin Hospital of Wuhan University, and written informed consent was obtained from each subject before any medical examination.Results:The fALFF values of DR patients in the left inferior temporal gyrus, left cerebellum, right pallidum, left supplementary motor area, and right superior orbitofrontal gyrus were -0.58±0.29, -0.47±0.39, -0.37±0.24, -0.31±0.26 and 0.29±0.49, respectively, which were significantly higher than -1.03±0.29, -0.90±0.31, -0.78±0.22, -0.72±0.21 and-0.17±0.46 of the normal control (voxel level P<0.01, gaussian random field correction, cluster level P<0.05)(all at P<0.001). Conclusions:Abnormal spontaneous neuronal activities occur in the left inferior temporal gyrus, left cerebellum, right pallidum, left supplementary motor area, and right superior orbitofrontal gyrus of DR patients.These abnormalities may associate with cognitive, vision and emotion.

Research Strategies and Key Problems Analysis over Substance Benchmark of Famous Classical Formulas / 中国实验方剂学杂志

With continuous introduction of relevant national policies on famous classical formulas, the research of famous classical formulas is popular all over the country. Different from other new drugs, in the research and development process of famous classical formulas, substance benchmark is earlier than the product, suggesting that the research and development of substance benchmark is of great significance. Based on previous work of the authors, content of substance benchmark of famous classical formulas was analyzed, which was included in the document <italic>The Management Regulation of Simplified Registration and Approval over Chinese Herbal Medicine Compound Preparations of Ancient Famous Classical Formulas</italic> released by the National Medical Products Administration in May 2018. In this paper, the significance of substance benchmark development was described, a five-stage of research strategy was proposed, covering the prescription textual research and historical evolution, the collection and quality evaluation of medicinal materials, the processing method and quality evaluation of decoction pieces, the preparation and quality research of substance benchmark, the drafting and formulating of quality standard over substance benchmark. At the same time, some suggestions were put forward to the feasibility of compound preparations development over famous classical formulas, the implementation difficulty of resource evaluation over Chinese medicinal materials, and the irrationality on the quality correlation of Chinese medicinal materials. All of these are expected to provide reference and enlightenment for the development and policy officially landed over ancient famous classical formulas.

Automated assisted clinical diagnosis of retinopathy of prematurity based on deep learning / 中华实验眼科杂志

Objective To evaluate the application value of an intelligent fundus assisted diagnosis system for detecting retinopathy of prematurity ( ROP) based on deep learning. Methods A total of 38895 fundus images for premature infants screening were collected from Renmin Hospital of Wuhan University Eye Center and were labeled by 10 licensed ophthalmologists. A deep learning network model was established to acquire automatic classification of disease stages and plus disease. The accuracy,sensitivity and specificity of the algorithm were calculated to evaluate the performance of the artificial intelligence system for ROP automatic diagnosis. This study protocol was approved by Ethic Committee of Renmin Hospital of Wuhan University ( No. WDRY2019-K032 ) . Written informed consent was obtained from the guardians of the children before entering the study cohort. Results The intelligent system achieved an accuracy of 0. 931. Specifically,the accuracies in detecting demarcation line (stageⅠ) was 0. 876,ridge (stage Ⅱ) was 0. 942,ridge with extra retinal fibrovascular (stageⅢ) was 0. 968,subtotal retinal detachment (stageⅣ) was 0. 998,total retinal detachment (stage Ⅴ) was 0. 999,vascular tortuosity and dilatation (plus disease) was 0. 896,optic disc was 0. 954,macular was 0. 781,and laser scars were 0. 974,respectively. Conclusions Deep learning algorithm can detect the stages and plus disease of ROP with excellent accuracy,and it provides the feasibility of applying the algorithm for ROP automated screening in clinical.