Objective To investigate the role of TROP2 in migration and invasion of human gastric cancer cells. Methods Small interfering RNA(siRNA) targeting TROP2 gene was constructed by gene cloning and transfection into gastric cancer cell line BGC-823. The expression of mRNA and protein were detected by Real-time quantitative PCR and Western blot assay after RNA interference. The proliferation was determined by MTT assay. Transwell assay was performed to assess the effect of TROP2 targeted RNA interference on the migratory and invasive properties of gastric cancer in vitro. Results Enzyme digestion analysis and DNA sequencing showed that TROP2 targeted RNA interference recombinant plasmids were successfully constructed. The most effective recombinant plasmid was selected. After transfection, knockdown of TROP2 significantly inhibited the proliferation, migration and invasion of BGC-823 cells in vitro(P <0. 05). Conclusion Interfering and down-regulating TROP2 gene can inhibit migration and invasion of gastric cancer cell line BGC-823 in vitro, indicating that TROP2 gene is a potential target for gastric cancer gene therapy.

Objective To analyze the mutation distribution of phenylalanine hydroxylase(PAH)gene in pa-tients with phenylketonuria(PKU)in Ningxia,and understand the hot spots and regions of PAH gene. Methods First-ly,6 hot mutation exons including 3,5,6,7,11,12 and their surrounding introns of the PAH gene in 30 patients with PKU in Ningxia were directly sequenced. And then the last 7 exons of 1,2,4,8,9,10,13 were sequenced for the patients in which 2 mutant alleles were not characterized. Multiplex ligation - dependent probe amplification (MLPA)was performed for the identification of uncharacterized mutant alleles after PAH sequence analysis of patients with PKU. Results Among 60 alleles,there were 58 mutant alleles(96. 7% ). Forty - six(81. 6% )mutant alleles were found in the exons 3,5,6,7,11,12. A total of 23 various mutations were detected,including missense(n = 9), splicing(n = 9),nonsense(n = 2),small deletion(n = 2)and large deletion(n = 1). The most common mutations were R243Q(18. 3% ),IVS4 - 1G ﹥ A(11. 7% )and R111X(11. 7% ). Among them,a novel mutation N393del was detec-ted in exon 11. MLPA identified a large deletion(c. - 1932 + 3402del)in 3 patients,1 of them was homoallelic muta-tion,and the others were heteroallelic mutation. Conclusions There are obvious hotspots and hot spot areas of PAH gene in PKU patients in Ningxia. There is a large deletion mutation in PKU patients in Ningxia. The MLPA is an effec-tive assay to detect large deletion in PAH gene.

Objective:To combine human anatomy preferably with Tibetan medical anatomy,thus,teaching quality and level of human anatomy has been improved.Method:To explain the major terms of Tibetan medical anatomy;combining with the Tibetan medical anatomy,to write the human anatomy teaching materials that fit the anatomy teaching of the Tibetan medical specialty to improve the teaching method;at the same time,to observe the teaching effects through the analyses of the questionnaire and exam results.Result:Shown as the investigation,those who think that the teaching materials have helped more to study human anatomy are taking 90.6%.After using this teaching material,people who think that those teaching materials has helped more tostudy Tisetan medical anatomy are taking 79.2%;Examination result analysis shows that,in two classes,before the teaching reform was implemented,students of above 80 marks were 12.5%,but after the teaching reform was implemented,those were 57.8%.Conclusion:The teaching material construction and teaching method reform brought into positive effects in the anatomy teaching quality improvement of the Tibetan medical specialty.

OBJECTIVETo observe the effect of Shouwu-Huanjing Recipe(SWHJR) medicated serum on human sperm motility and fertility in vitro.

METHODSHuman sperm was co-cultured with SWHJR medicated serum in vitro. Human sperm motility was evaluated by computer-assisted semen analysis(CASA). The acrosome reaction and the capability of penetrating zona-free hamster eggs were also observed.

RESULTSThe co-cultured SWHJR medicated serum significantly increased the sperm motion velocity(VAP, VCL, VSL) (P < 0.01), the amplitude of lateral head movement (ALH) and the beat frequency of flagellum(BCF), the density of progressive motility sperms (P < 0.05), the acrosome reaction rate(P < 0.001), the fertilization rate(FR) and the fertilization index (FI) in sperm penetration assay(SPA) test (P < 0.01). The stimulation of SWHJR medicated serum occurred in dose-dependent manner.

CONCLUSIONSWHJR can improve human sperm motility and fertility.

Acrosome Reaction ; Animals ; Cricetinae ; Fertility ; drug effects ; Humans ; Male ; Medicine, Chinese Traditional ; Mesocricetus ; Rats ; Rats, Sprague-Dawley ; Sperm Motility ; drug effects

Objective: To explore the protective effects of folic acid on retinas and its anti-oxidative stress mechanism in diabetic mice. Methods: Thirty-two 16-week-old SPF degree male db/db mice were randomized into model group and folic acid group, and 16 matched C57BL/KsJ mice were used as controls.Folic acid was used to the mice by oral gavage once per day with the dose of 71 μg/kg (2 ml) for 60 days in the folic acid group, and the same volume of normal saline solution was used in the model group and control group in the same way.The activities, mental state, body weight, and fasting plasma glucose (FPG) of the mice were recorded during experiment.At the end of the intervention, the mice were sacrificed and the retinas and blood sample were obtained.The histopathology of the retinas was examined with hematoxylin- eosin staining; serum homocysteine (Hcy) was detected by ELISA assay; the relative expressions of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were detected in the retinas by real-time fluorescence quantitative PCR; the relative expressions of B lymphoma 2 protein (bcl-2), bcl-2 related X protein (bax), 3-nitrotyrosine (3-NT) and 4-Hydroxynonine (4-HNE) proteins were assayed by Western blot assay; superoxide dismutase (SOD), 8-hydroxydeoxyguanosine (8-OHdG) and malondialdehyde (MDA) levels in the retinas were detected by biochemical kits, and immunofluorescence assay was used to detect the expression of NADPH oxidation 4 (NOX4) in the retinas.The use and care of the experimental animals adhered to the ARVO Statement by the American Association for Vision and Ophthalmology Research and this study protocol was approved by Ethic Committee of Qinghai University (QHDX 2018-35). Results: Over the experimental period, The FPG was normal and body weight was gradually increased in the mice of control group.The FPG>16.7 mmol/L and the mice appeared obese.In the folic acid group, both body mass and FPG of the mice were gradually reduced.At the end of drug administration, serum Hcy concentration of the mice was (27.18±3.18)μmol/L in the model group, which was significantly higher than (8.28±2.18)μmol/L in the control group and (13.73±2.54)μmol/L in the folic acid group (all at P<0.05). The retinal structure was intact in the control group, and the retinas were thinning with more capillaries and inflammatory cells in the model group, the thickness of the retinas was increased and the capillaries and inflammatory cells were decreased in the folic acid group.The relative expressions of TNF-α and IL-6 mRNA in the retinas were significantly higher in the model group than those in the control group, and those in the folic acid group were reduced in comparison with the model group (all at P<0.05). The relative expression of bcl-2 protein in the retinas of folic acid group was lower than that in the control group and higher than that in the model group, and the relative expressions of bax, 3-NT and 4-HNE proteins in the retinas of the folic acid group were significantly higher than those in the control group and lower than those in the model group (all at P<0.05 ). The T-SOD activity in the folic acid group was significantly stronger than that in the control group and weaker than that in the model group, and the concentrations of 8-OHdG and MDA in the retinas of the folic acid group were significantly reduced in comparison with those of the control group and elevated in comparison with those of the model group (all at P<0.05). The expressing intensity of NOX4 protein in the retinas of the folic acid group was significantly weaker than that of the model group. Conclusions Folic acid appears aprotective effect on retinal tissue in diabetic mice by reducing serum Hcy, inhibiting oxidative stress and cell apoptosis.

ObjectiveTo explore the mechanism of Tibetan medicine Ershiwuwei Guijiuwan against osteoporosis in ovariectomized rats through the classical Wnt/β-catenin signaling pathway. MethodForty-eight 3-month-old SD female rats were randomized into model group （equivalent volume of normal saline）， sham operation group （equivalent volume of normal saline）， estradiol group （0.1 mg·kg^-1 estradiol valerate）， and high-， medium-， low-dose Ershiwuwei Guijiuwan groups （800， 400， 200 mg·kg^-1， respectively）. For the modeling， some adipose tissue near the ovaries was removed in the sham operation group， and ovaries were excised in other groups. Administration （ig， once a day） started two weeks after the operation and lasted 12 weeks. After sampling， based on hematoxylin and eosin （HE） staining， the morphological changes of the right femur and lumbar spine of the rats were observed. The enzyme-linked immunosorbent assay （ELISA） was performed to detect the serum levels of rat tartrate-resistant acid phosphatase 5b （TRAP5b）， collagen type Ⅰ C-terminal peptide （CTX-Ⅰ）， bone alkaline phosphatase （BALP）， amino-terminal propeptide of type Ⅰ procollagen （PINP）， and bone Gla-protein （BGP）. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was performed to examine the mRNA expression of β-catenin and Runt-related transcription factor 2 （Runx2） in femur， and Western blot to detect the protein expression of β-catenin， Runx2， and low-density lipoprotein receptor-related protein-5 （Lrp-5）. ResultCompared with the sham operation group， the model group showed disordered and sparse bone trabecula with fewer connections， decrease in serum levels of BALP， BGP， and PINP （P<0.01）， increase in levels of CTX-Ⅰ and TRAP5b （P<0.01）， reduction in mRNA expression of β-catenin and Runx2 in femoral tissue （P<0.01） and protein expression of Lrp-5， β-catenin， and Runx2 （P<0.01）. Compared with the model group， estradiol and each dose of Ershiwuwei Guijiuwan increased the volume of bone trabecula， made thebone trabecula closely arranged， reduced the loss of the trabecular meshwork， raised the serum levels of BALP and BGP （P<0.01）， and lowered TRAP5b level （P<0.01）. PINP level was significantly increased in the high-dose Ershiwuwei Guijiuwan group and estradiol group （P<0.01） and CTX-Ⅰ level was significantly decreased in the high-dose Erwenwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. The mRNA expression of β-catenin in femoral tissue and protein expression of Lrp-5 and β-catenin in estradiol group and three Ershiwuwei Guijiuwan groups were significantly increased （P<0.05， P<0.01）， and the levels of Runx2 mRNA and protein were significantly increased in the high-dose Ershiwuwei Guijiuwan group and the estradiol group （P<0.01） compared with those in the model group. ConclusionTibetan medicine Ershiwuwei Guijiuwan is effective for the osteoporosis in ovariectomized rats， which may be related to the classic Wnt/β-catenin signaling pathway. It affects bone metabolism by up-regulating the expression of osteogenesis-related genes in the signaling pathway.

To analyze the use of outcome indicators of randomized controlled trial(RCT) of acupuncture in the treatment of acute ischemic stroke in recent three years, so as to provide a basis for building a study on the core outcome indicators for the treatment of acute ischemic stroke with acupuncture. The RCTs of acupuncture treatment for acute ischemic stroke in recent three years were collec-ted through computer retrieval of eight Chinese and English databases and two clinical trial registries at home and abroad. Literature was screened out, and data was extracted. Risk of assessment bias tool Cochrane 6.1 was used for bias risk assessment, outcome indicators were summarized and analyzed. A total of 47 RCTs were included, and 3 studies were trials registration scheme. Outcome indicators were divided into 6 categories according to functional attributes, namely physical symptoms/signs, physical and chemical examination, quality of life, traditional Chinese medicine symptoms/syndromes, safety events and long-term prognosis. The study found that in addition to the common problems in previous studies covered by the status quo of outcome indicators selection of RCT of acupuncture in the treatment of acute ischemic stroke, there were also the other problems as follows: emphasis on macroscopic efficacy indicators but neglect of acupuncture specific indicators, lack of characteristic indicators and economic indicators of traditional Chinese medicine therapy, and unification of indicators measurement tool and measurement time point. In the future, the outcome indicators set for the treatment of acute ischemic stroke with acupuncture shall be established, and the core outcome indicators set shall be in line with the characteristics of traditional Chinese medicine treatment.
Acupuncture Therapy ; Brain Ischemia/therapy* ; Humans ; Ischemic Stroke ; Quality of Life ; Randomized Controlled Trials as Topic ; Stroke/therapy* ; Treatment Outcome

There have been many clinical trials, systematic reviews/Meta-analysis proving that Xingnaojing Injection has a good clinical efficacy in treatment of cerebral ischaemic stroke, but with fewer comprehensive descriptions. In this study, an overview of systematic reviews/Meta-analysis of Xingnaojing Injection in treating cerebral ischaemic stroke was performed to provide current situation of evidences and basis for clinical practice. CNKI, Wanfang, VIP, CBM, EMbase, PubMed, Cochrane Library, Web of Science were retrieved through computers. A total of 6 literatures were included in this study. By AMSTAR-2 checklist and GRADE, the quality of included systematic reviews and the efficacy of Xingnaojing Injection were evaluated. The results of AMSTAR-2 checklist showed an extremely low quality for all of the 6 systematic reviews. According to the results of GRADE evaluation, among 55 outcomes, there were 2 outcomes with a medium quality, 4 outcomes with a low quality and 49 outcomes with an extremely low quality. The 6 systematic reviews reached a consistent conclusion that Xingnaojing Injection was effective in the treatment of cerebral ischaemic stroke. This therapy could improve the total efficacy, neurological deficit scores, hemodynamic and hemodynamic parameters. However, the methodolo-gical quality of all literatures was extremely low. The evidence levels of outcomes were between extremely low to medium. The effectiveness of Xingnaojing Injection in the treatment of cerebral ischaemic stroke still needs to be further verified by more high-quality studies. In the future, relevant clinical studies and systematic reviews/Meta-analysis shall be carried out in a strict accordance with relevant regulations.
Brain Ischemia/drug therapy* ; Drugs, Chinese Herbal ; Humans ; Ischemic Stroke ; Stroke/drug therapy* ; Systematic Reviews as Topic

To analyze the efficacy and safety of Shensong Yangxin Capsules in treatment of bradycardia combined with premature beat. Databases, such as CNKI, VIP, WanFang, SinoMed, PubMed, Cochrane Library, ClinicalTrials were retrieved by computers for relevant randomized controlled trials of Shensong Yangxin Capsules in treatment of bradycardia combined with premature beat. Two researchers independently screened out the literatures, extracted data according to the inclusion criteria, and applied the Risk of Bias assessment tool in assessing the methodological quality. The Cochrane systematic evaluation software RevMan 5.3 was used for data analysis. Totally 9 randomized controlled trials including 706 subjects were included. The intervention measure was the single administration with Shensong Yangxin Capsules, and the control measure was the blank control. The results showed that Shensong Yangxin Capsules had an obvious effect on average heart rate(MD=6.59, 95%CI[3.87, 9.31], I~2=90%), premature beat efficacy(RR=1.72, 95%CI[1.53, 1.93], I~2=0%), heart rate efficacy(RR=1.74, 95%CI[1.40, 2.17], I~2=47%), and objective efficacy(RR=1.50, 95%CI[1.31, 1.70], I~2=31%). Eight studies reported safety events, with no significant adverse reaction. In conclusion, the single administration with Shensong Yangxin Capsules may have a certain effect in improving heart rate, controlling premature beats and alleviating clinical symptoms in patients with bradycardia combined with premature beat, with no obvious adverse reaction. Shensong Yangxin Capsules can be used in clinic. This potential conclusion needs to be confirmed in future trials using rigorous methodology.
Bradycardia/drug therapy* ; Capsules ; Cardiac Complexes, Premature/drug therapy* ; Drugs, Chinese Herbal/therapeutic use* ; Humans ; Randomized Controlled Trials as Topic

Aim To investigate the effects of combined administration of loganin and berberine on bone structure and metabolism in diabetic mice and its potential mechanism.Methods The diabetic ICR mouse model was induced by high fat diet(HFD).After 10 weeks of combined intervention, the effects of loganin and berberine on body weight, body fat rate, blood glucose, blood lipid and serum oxidative stress levels were observed.Bone microstructure was scanned by micro-CT.Biomechanical characteristics of bone were measured by three-point bending test, and material properties were detected by fourier transform infrared(FTIR).The pathological changes were observed by HE and TRAP staining.Protein expressions involved in advanced glycation end products(AGEs)and their receptors(RAGE)/nuclear factor-κB(NF-κB)signaling pathway were detected by immunohistochemistry.Results The combined administration of loganin and berberine could significantly inhibit the weight gain, reduce the levels of blood glucose, blood lipid and oxidative stress, as well as improve glucose tolerance.In addition, combined intervention also decreased the expression levels of the proteins involved in AGEs/RAGE/NF-κB signaling pathway, and improved bone microstructure, finally contributing to increasing bone quality in diabetic mice.Conclusions The combination of loganin and berberine could improve bone metabolism in diabetic mice, which may be related to AGEs/RAGE/NF-κB signaling pathway.