Principles and strategies for species identification based on analysis of whole-genome / 药学学报

The main sources of natural drugs include various biological species such as plants, animals, and microorganisms. The accurate identification of these species is the bedrock of natural drug development. We propose a novel method of species identification in this paper: analysis of whole-genome (AGE), a molecular diagnostic method used to identify species by finding species-specific sequences from the whole genome and precisely recognizing the specific target sequences. We elaborate that the principle for species identification based on AGE is that the genome sequences of diverse species must differ and divide the implementation strategy of the method into two levels of research and application. Based on our analysis of its characteristics, the method would have the potential advantages of reliable principle, high specificity, and wide applicability. Moreover, three crucial concerns related to building method systems including genome acquisition, bioinformatics analysis, and database construction, are further discussed. In summary, we offer theoretical underpinnings and methodological guidance for the development of bioinformatics software and commercial kits, indicating AGE has great application potential in objects, subjects, and industries.

Application of DNA barcoding technology to national drug sampling inspection / 药学学报

Adulterants and counterfeits were found in some of the commercial traditional Chinese medicine (TCM) decoctions in Hongjin Xiaojie Jiaonang, Hongjin Xiaojie Pian, and Chaihuang Keli during the national drug sampling inspection. However, it was difficult to determine the species of the adulterants and counterfeits by conventional testing methods. Therefore, a total of 184 samples of the TCM decoctions and raw materials belong to the prescriptions of above mentioned traditional Chinese patent medicines, including Bupleuri Radix, Bajiaolian, Heimayi, and Shufuchong, were collected and authenticated by DNA barcoding technology. 111 ITS2 sequences were obtained from 115 commercial TCM decoctions and raw materials of Bupleuri Radix, among which 71 were Bupleurum chinense, three were B. scorzonerifolium, and 31 were closely related species in the same genus. In addition, counterfeits derived from different genera, such as Ailanthus altissima (one sample), Saposhnikovia divaricate (two samples), and Solidago decurrens (three samples), were also detected. 21 ITS2 sequences were obtained from 22 commercial TCM raw materials of Bajiaolian, among which 15 were Diphylleia sinensis and six were Dysosma versipellis and other species in genus Dysosma. For 22 Heimayi samples, PCR amplification of COI sequence was failed due to genomic DNA degradation. Among 38 Shufuchong samples, 24 COI sequences were obtained and only nine of them were the genuine species (Armadillidium vulgare) recorded in the Chinese Pharmacopoeia, 11 were Porcellio laevis, two were Mongoloniscus sinensis, and two samples could not be identified due to the limitation of database. This study demonstrates that DNA barcoding technology is suitable for the species authentication of the decoctions of traditional Chinese patent medicine prescription. It is a conductive way for the establishment of traceability system for the whole TCM industrial chain.

DNA barcoding identification of commercial decoctions in traditional Chinese medicine / 药学学报

Although the guiding principles for molecular identification of traditional Chinese medicines (TCM) using DNA barcoding have been recorded in the Chinese Pharmacopoeia, there is still a lack of systematic research on its application to commercial TCM decoctions. In this study, a total of 212 commercial TCM decoctions derived from different medicinal parts such as root and rhizome, fruit and seed, herb, flower, leaf, cortex, and caulis were collected to verify applicability and accuracy of the method. DNA barcodes were successfully obtained from 75.9% (161/212) of the samples, while other samples failed to be amplified due to genomic DNA degradation. Among the 161 samples, 85.7% of them were identified as recorded species in the Chinese Pharmacopoeia (2020 edition). In addition, 14 samples could be identified as species recorded in the Chinese Pharmacopoeia and their closely related species in the same genus. Morphological identification for the unconfirmed samples showed that eight were genuine species and three were adulterants, while the other three were unidentifiable due to lack of morphological characteristics. Furthermore, the DNA barcodes of seven samples accurately mapped to the sequences of adulterants. Remarkably, counterfeit products were detected in two samples. These results demonstrate that DNA barcoding is suitable for the identification of commercial TCM decoctions. The method can effectively detect adulterants and is appropriate for use throughout the industrial chain of TCM production and distribution, and by the supervisory agencies as well.

Therapeutic effect of herb-separated moxibustion at Jinsuo (GV 8)-eight-diagram points on diarrhea-type irritable bowel syndrome of liver stagnation and spleen deficiency / 中国针灸

OBJECTIVE@#To observe the clinical therapeutic effect of herb-separated moxibustion at Jinsuo (GV 8)- eight-diagram points on diarrhea-type irritable bowel syndrome (IBS-D) of liver stagnation and spleen deficiency as compared with oral administration of pinaverium bromide tablets and Chinese herbal decoction, .@*METHODS@#A total of 126 patients with IBS-D of liver stagnation and spleen deficiency were randomized into a herb-separated moxibustion group (moxibustion group), a western medication group and a Chinese herbal medication group, 42 cases in each one. In the moxibustion group, the herb-separated moxibustion was applied to Jinsuo (GV 8)-eight-diagram points. The herbs in (fried , fried , and ) were ground into herbal paste and the paste was put on Jinsuo (GV 8)-eight-diagram points. The suspending moxibustion was exerted over the points for 40 min, once daily. In the western medication group, pinaverium bromide tablets were taken orally, 50 mg each time, three times a day. In the Chinese herbal medication group, the decoction of was taken orally, one dose a day, taking separately in two times. The duration of treatment was 8 weeks in each group. Before and after treatment, the symptom score of traditional Chinese medicine (TCM), gastrointestinal (GI) symptom score, the score of IBS symptom severity scale (IBS-SSS) and the score of IBS quality of life (IBS-QOL) scale were observed in patients of each group separately. The clinical therapeutic effect was evaluated.@*RESULTS@#After treatment, the scores of TCM symptoms, GI symptom scores and IBS-SSS scores were all obviously reduced in each group (<0.05). Each of the scores in the moxibustion group was lower than the western medication group and the Chinese herbal medication group respectively (<0.05). After treatment, the scores of each of eight subscale structures of IBS-QOL scale, named dysphoria, interference with activity, body image, health worry, food avoidance, social reaction, sexual intercourse and relationship, were all increased obviously in each group (<0.05). The scores of each of eight subscale structures in the moxibustion group were higher than the western medication group and the Chinese herbal medication group respectively (<0.05). The total effective rate was 92.9% (39/42) in the moxibustion group, higher than 71.4% (30/42) in the western medication group and 73.8% (31/42) in the Chinese herbal medication group respectively (<0.05).@*CONCLUSION@#Herb-separated moxibustion at Jinsuo (GV 8)-eight-diagram points remarkably relieves gastrointestinal symptoms and improves the quality of life in patients of diarrhea-type irritable bowel syndrome of liver stagnation and spleen deficiency, and its clinical therapeutic effect is superior to oral administration of either pinaverium bromide tablets or .

Production and quality control of original herbal materials of Danhong Injection / 中国中药杂志

Cardiovascular and cerebrovascular diseases are the leading cause of death for residents in China. Danhong Injection(DHI) decoction piece is prepared from Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos, with the function of promoting the blood circulation, removing the blood stasis, relaxing the sinews and dredging the collaterals. In recent years, about 100 million bottles of DHI have been sold. Consequently, its safety and effectiveness are very important to a large number of patients. Raw materials are the source and foundation for production of traditional Chinese medicine injections. In this article, we reviewed the identification of Salviae Miltiorrhizae Radix et Rhizoma and Carthami Flos, resource distribution, cultivation, quality control, and detection of xenobiotic pollutants, in order to guide the production of high-quality, stable, and pollution-free raw materials. This will be a benefit in ensuring the safety and effectiveness of DHI and reducing the incidence of adverse reactions from the raw materials. By comparing the similarities and differences between the quality standards of Salviae Miltiorrhizae Radix et Rhizoma, Carthami Flos and DHI, we provided some comments for improving the quality standards and post-marketing reevaluation of DHI, and provided some theoretical supports for the production of high-quality herbal raw materials.

Influence of general anesthetic exposure in developing brain on cognition and the underlying mechanisms / 生理学报

With the evolution of medical techniques and technology, an increasing number of infants, neonates, and fetuses are exposed to general anesthesia for clinical diagnostic and therapeutic process. The neurotoxic effects of general anesthetics on developing brain have been a subject of concern and considerable research interest. Population-based study confirmed that single short-term general anesthetic exposure does not affect nervous system function, but multiple exposures to general anesthesia could damage cognitive function. Animal studies further discovered the underlying mechanisms. Nervous system is most susceptible to general anesthetics during the brain growth spurt. The time-point is more critical than the duration of exposure to general anesthetics. General anesthetics can induce intracellular calcium overload, disturb energy metabolism, promote cell apoptosis and lead to cell loss. General anesthetics can damage synaptic structure, transmission and plasticity, and impair brain function. High throughput omics technologies have been used to screen the differentially expressed genes induced by general anesthetics, which provide further understanding of the mechanism of general anesthetics affecting cognitive function. This review provides an update on the pathophysiologic mechanisms underlying the anesthesia-neurotoxicity, which will be helpful to provide instructions for the clinical use of general anesthesia in children.

Effect of eicosapentaenoic acid on mRNA expression of tight junction protein ZO-1 in intestinal epithelial cells after Escherichia coli LF82 infection / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To investigate the change in the expression of tight junction protein ZO-1 in intestinal epithelial cells (Caco-2 cells) and the protective effect of eicosapentaenoic acid (EPA) after adherent-invasive Escherichia coli (E.coli) LF82 infection.</p><p><b>METHODS</b>The Caco-2 cell line was used to establish an in vitro model of tight junction of intestinal epithelial cells. Caco-2 cells were divided into EPA treatment groups (0, 25, 50, 100, and 200 μmol/L EPA) and EPA (0, 25, 50, 100, and 200 μmol/L EPA)+E.coli LF82 treatment (0, 6, and 12 hours) groups. A microscope was used to observe the morphological characteristics of the cells. MTT assay was used to determine the cell growth curve. The activity of alkaline phosphatase (ALP) at both sides of the cell membrane was compared to evaluate the Caco-2 cell model. MTT assay and flow cytometry were used to investigate the effects of different concentrations of EPA on the survival rate and apoptosis rate of Caco-2 cells. RT-qPCR was used to measure the mRNA expression of ZO-1 in Caco-2 cells after EPA and/or E.coli LF82 treatment. ELISA was used to measure the change in the level of tumor necrosis factor-α (TNF-α) in culture supernatant.</p><p><b>RESULTS</b>After EPA treatment (25 and 50 μmol/L), the proliferation of Caco-2 cells was induced in a dose-dependent manner. The survival rates of the cells were significantly higher than those in the control group (P<0.05). The EPA treatment (100 and 200 μmol/L) groups had a significant inhibitory effect on the proliferation of Caco-2 cells in a dose-dependent manner. The survival rates of the cells were significantly lower than those in the control group (P<0.05). The EPA treatment (100 and 200 μmol/L) groups had a significant increase in cell apoptosis rate compared with the control group (P<0.05). The 6- and 12-hour E.coli LF82 treatment groups had decreasing mRNA expression of ZO-1 in Caco-2 cells over the time of treatment and had significantly lower mRNA expression of ZO-1 than the untreated group (P<0.05). The Caco-2 cells treated with E.coli LF82 and 25 or 50 μmol/L EPA for 6 or 12 hours showed an increase in the mRNA expression of ZO-1 with the increasing concentration of EPA, as well as significantly higher mRNA expression of ZO-1 than the Caco-2 cells treated with E.coli LF82 alone (P<0.05). The Caco-2 cells treated with E.coli LF82 alone for 6 or 12 hours had increasing secretion of TNF-α over the time of treatment and had significantly higher secretion than the untreated Caco-2 cells (P<0.05). The Caco-2 cells treated with E.coli LF82 and 25 or 50 μmol/L EPA for 6 or 12 hours showed a reduction in the secretion of TNF-α with the increasing concentration of EPA and had significantly lower secretion than the Caco-2 cells treated with E.coli LF82 alone (P<0.05).</p><p><b>CONCLUSIONS</b>EPA can effectively prevent the destruction of tight junction of intestinal epithelial cells induced by E.coli LF82 infection and inhibit the secretion of inflammatory factors. Therefore, it has a certain protective effect on intestinal mucosal barrier.</p>

Protective effect of omeprazole on gastric mucosal of cirrhotic portal hypertension rats

OBJECTIVE@#To observe the protective effect of omeprazole on gastric mucosal of cirrhotic portal hypertension rats.@*METHODS@#All rats were randomly divided into normal control group, cirrhosis and treatment group. Thioacetamide was used to establish rat model of cirrhotic portal hypertension. The necrotic tissue of gastric mucosa ulcer focus, degree of neutrophils infiltration at the ulcer margin, portal pressure, portal venous flow, abdominal aortic pressure, abdominal aortic blood flow at front end, gastric mucosal blood flow (GMBF), glycoprotein (GP) of gastric mucosa, basal acid secretion, H(+)back -diffusion, gastric mucosal damage index, NO, prostaglandin E2(PGE2) and tumor necrosis factor-α (TNF-α) were determined respectively, and the pathological changes of gastric mucosa were also observed by microscope.@*RESULTS@#Compared with cirrhosis group and the control group, the ulcer bottom necrotic material, gastric neutrophil infiltration and UI of the treatment group were all decreased significantly (P<0.01), GMBF value, GP values, serum NO, PGE2, TNF-α were all significantly increased.@*CONCLUSIONS@#Omeprazole has an important protective effect on gastric mucosal and it can increase gastric mucosal blood flow and related to many factors.

Biodistribution and preparation of technetium-99m-labeled D-D₃ monoclonal antibody against pro-gastrin-releasing peptide (₃₁₋₉₈) in mice / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>We previously reported that iodine-131((131)I)-labeled anti-pro-gastrin-releasing peptide (ProGRP(31-98)) monoclonal antibody D-D3 could selectively accumulate in the tumor sites of nude mice bearing small cell lung cancer (SCLC) xenografts. However, (131)I-D-D3 was cleared slowly from the body, and the best radioimmunoimaging time for SCLC was 72 - 96 hours after injection. The aims of this study were to radiolabel anti-ProGRP(31-98) D-D3 monoclonal antibody with technetium-99m ((99m)Tc) and to investigate the biodistribution of this antibody in healthy ICR mice.</p><p><b>METHODS</b>D-D3 was labeled with (99m)Tc via the 2-mercaptoethanol reduction method. (99m)Tc-D-D3 was purified by the gel column separation method. The labeling efficiency and radiochemical purity were measured by thin-layer chromatography. The immunological activity of (99m)Tc-D-D3 was determined with cell conjugation assays. (99m)Tc-D-D3 was injected into healthy ICR mice via a tail vein, and all the healthy ICR mice were sacrificed by cervical dislocation at a designated time. Then, the blood and major organs were removed and weighed, and counted in a gamma scintillation counter to determine the percentage of the injected dose per gram (%ID/g).</p><p><b>RESULTS</b>The labeling rate and the radiochemical purity of (99m)Tc-D-D3 were (73.87 ± 2.89)% and (94.13 ± 4.49)%, respectively. The immunobinding rates of (99m)Tc-D-D3 to the human small cell lung cancer NCI-H446 cell line and lung adenocarcinoma A549 cell line were (81.2 ± 2.37)% and (24.3 ± 1.46)%, respectively. The distribution data of normal ICR mice demonstrated that (99m)Tc-D-D3 was mainly distributed in the liver, kidney and lung, and less in the brain tissue and muscle.</p><p><b>CONCLUSIONS</b>(99m)Tc-D-D3 antibody not only had high radiochemical purity, but also had good stability both in vitro and in vivo, and maintained good immunological activity. (99m)Tc-D-D3 was metabolized mainly in the kidney and liver, and the blood radioactivity decreased rapidly. Thus, (99m)Tc-D-D3 is conducive to the radioimmunoimaging of SCLC.</p>

Controlled release of transforming growth factor-beta receptor kinase inhibitor from thermosensitive Chitosan-based hydrogel: application for prevention of capsular contracture / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Capsular contracture has become the most common complication associated with breast implant. Transforming growth factor-beta (TGF-β) is well known for a prominent role in fibrotic diseases. Due to the critical role of TGF-β in pathogenesis of capsular formation, we utilized thermosensitive C/GP hydrogel to controlled release of TGF-β receptor kinase inhibitor (SD208) and investigated their effects on capsular contracture.</p><p><b>METHODS</b>In vitro degradation and drug release of C/GP hydrogel were performed. Twenty-four rabbits underwent subpanniculus implantation with 30 ml smooth silicone implants and were randomly divided into four groups as follows: Group 1 received saline solution; Group 2 received SD208; Group 3 received SD208-C/GP; Group 4 received C/GP. At 8 weeks, the samples of capsular tissues were analyzed by hematoxylin and eosin and immunohistological staining. The mRNA expression of collagen III and TGF-β1 was detected by RT-PCR assay.</p><p><b>RESULTS</b>C/GP hydrogel could be applied as an ideal drug delivery vehicle which supported the controlled release of SD208. SD208-C/GP treatment showed a significant reduction in capsule thickness with fewer vessels. The histological findings confirmed that the lower amounts of inflammatory cells and fibroblasts infiltrate in SD208-C/GP group. In contrast, typical capsules with more vessel predominance were developed in control group. We did not observe the same inhibitory effect of SD208 or C/GP treatment on capsular contracture. Moreover, SD208-C/GP therapy yielded an evident down-regulation of collagen III and TGF-β1 mRNA expression.</p><p><b>CONCLUSIONS</b>This study demonstrated that controlled release of TGF-β receptor kinase inhibitor from thermosensitive C/GP hydrogel could significantly prevent capsule formation after mammary implants.</p>

Protective effect of limb ischemic preconditioning against liver ischemia/reperfusion injury in the rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the protective effect of limb ischemic preconditioning against liver ischemia/reperfusion injury in the rat.</p><p><b>METHODS</b>Rats were randomly divided into four groups (n = 8): (1) Sham group (S group), rats without ischemic preconditioning (IPC), (2) Ischemia/reperfusion (I/R) without IP (I/R group); (3) Rats with 5 min IPC (IPC group); (4) Rats with lower limbs IPC and repeated three times (remote ischemic preconditioning, RPC group); The rats were subjected to 60-min sustained liver ischemia followed by 180-min reperfusion except S group. All ischemia rats were only subjected to 70% liver ischemia. Finally, blood and liver samples were obtained to determine the activity of ALT and AST, liver wet/dry weight (W/D), PMN counts and pathology.</p><p><b>RESULTS</b>All IPC group and RPC group had obviously lower levels of ALT, AST, W/D, PMN counts than that of the I/R group (P < 0.01).</p><p><b>CONCLUSION</b>The limb ischemic preconditioning has a protective effects against liver ischemia/reperfusion injury in the rat, possibly are due to suppression of liver inflammatory reaction, improvement of liver microcirculation.</p>

3D reconstruction based on sub-pixel points of structured light / 中国医疗器械杂志

In this paper, a method for detecting sub-pixel points based on zero-crossing detection combined with Steger's curvilinear detector has been presented. Finally, the feasibility and validity of this method has been confirmed by the experiment.

Corrlation of dorsal artery of foot with type 2 diabetes mellitus on ultrasonography / 中华医学超声杂志（电子版）

Objective To evaluate the correlation between lesions of dorsal artery of foot and type 2 diabetes on CDFI. Methods Dorsal artery of foot was examinated in 97 cases with type 2 diabetes and 46 cases without diabetes mellitus. Results There were variable changes in intima-media of dorsal artery of foot in type 2 diabetes patients.And the patients with hyperlipidemia and hypertension showed serious lesions in the dorsal artery of foot and bifurcation of the blood vessel .Simple arteriosclerosis showed not only lesions in intima-media of dorsal artery of foot but also relative mild lesions in bifurcation of the blood vessel. Blood vessel lesion incidence in type 2 diabetes was significantly higher than that of the control group (P<0.01). Conclusions There was a significant correlation between lesions of dorsal artery of foot and type 2 diabetes patient's condition. CDFI is an effective method in evaluating patient's condition, degree and prognosis of type 2 diabetes, and has an important clinical value in early diagnosis, prognosis and treatment.Simple arteriosclerosis showed focal lesions in bifurcation of the blood vessel while dorsal artery of foot showed relatively mild lesions .

Low positive AFP for the diagnosis of hepatocellular carcinoma / 中国基层医药

Objective To investigate the value of low positive AFP level for the diagnosis of hepatocellular carcinoma(HCC)when a space occuping lesion was already identified in the liver.Methods The AFP level of 401 HCC cases from January 1999 to October 2006 were retrospectively analyzed.Results 22 cases of small liver cancer underwent reducing surgical resection in our hospital,increasing the quality of life and prolonging survival rate.Con- clusion(1)The AFP level between 20 and 200?g/L is of diagnostic for HCC when a SOL is identified.(2)The clinical reference diagnostic criteria using AFP level more than 20?g/L,instead of more than 200?g/L is helpful for the diagnosis.(3)Combined diagnosis of AFP level more than 20?g/L and ultrasonography or CT scanning yields higher sensitivity and specificity th.an traditional,diagnostic criteria.

Advances in the studies of Kallmann syndrome / 中华男科学杂志

Kallmann syndrome (KS) is a rare hereditary disease. It is characterized by hypogonadotrophic hypogonadism in association with anosmia or hyposmia. At present, three modes of inheritance and genes related to KS have been identified. This review focuses on the clinical diagnosis and advances in the studies of the pathogenesis gene for Kallmann syndrome.

Analysis of an AZFc deletion family with natural transmission / 中华男科学杂志

<p><b>OBJECTIVE</b>To report a rare family of AZFc deletion with natural transmission and explore the potential mechanism by which identical microdeletions cause different phenotypes.</p><p><b>METHODS</b>Chromosomal quantity and construction were detected by G-band, Y-chromosomal microdeletions by multiple PCR amplification for 12 sequence tagged sites (STSs, and the single-nucleotide polymorphisms (SNPs) of the DAZL gene, the autosomal homologue of deleted-in-azoospermia (DAZ) gene by DNA sequencing.</p><p><b>RESULTS</b>Chromosome analysis revealed a normal karyotype 46, XY in the father and both of his two sons and microdeletions of the full AZFc region were identical, including sY152, sY157, sY242, sY254, sY255, sY239 locus. However, the phenotypes of the affected patients were different: the father had normal fertility, but the sperm density of his two sons deteriorated age-dependently, and the younger one suffered from left cryptorchidism. SNP analysis demonstrated that two polymorphisms in exon 2 and 3 of the DAZL gene were identical in both the father and his sons.</p><p><b>CONCLUSION</b>Identical Y-chromosomal microdeletions causing different phenotypes in this family is not associated with the polymorphisms of DAZL gene and may be related to other genes or environmental factors.</p>

Diagnosis and differential diagnosis of Kallmann syndrome / 中华男科学杂志

<p><b>OBJECTIVE</b>To study diagnosis and differential diagnosis of Kallmann syndrome.</p><p><b>METHODS</b>The examinations including routine karyotyping, sex hormone, GnRH stimulation test and MRI were performed.</p><p><b>RESULTS</b>Cytogenetic analysis of his peripheral lymphocyte by G banding showed a normal male karyotype. GnRH stimulation test presented a good reaction. Plasma levels of luteinizing hormone (LH), follicle-stimulating hormone (FSH) and testosterone were very low. Absent olfactory bulb was found by magnetic resonance imaging (MRI).</p><p><b>CONCLUSION</b>Karyotype analysis, sexual hormone, GnRH stimulation test and MRI are very important the diagnosis of Kallmann syndrome.</p>