Unexpected mutations occurred in CRISPR/Cas9 edited Drosophila analyzed by deeply whole genomic sequencing.

CRISPR/Cas9 possesses the most promising prospects as a gene-editing tool in post-genomic researches. It becomes an epoch-marking technique for the features of speed and convenience of genomic modification. However, it is still unclear whether CRISPR/Cas9 gene editing can cause irreversible damage to the genome. In this study, we successfully knocked out the WHITE gene in Drosophila, which governs eye color, utilizing CRISPR/Cas9 technology. Subsequently, we conducted high-throughput sequencing to assess the impact of this editing process on the stability of the entire genomic profile. The results revealed the presence of numerous unexpected mutations in the Drosophila genome, including 630 SNVs (Single Nucleotide Variants), 525 Indels (Insertion and Deletion) and 425 MSIs (microsatellite instability). Although the KO (knockout) specifically occurred on chromosome X, the majority of mutations were observed on chromosome 3, indicating that this effect is genome-wide and associated with the spatial structure between chromosomes, rather than being solely limited to the location of the KO gene. It is worth noting that most of the mutations occurred in the intergenic and intron regions, without exerting any significant on the function or healthy of the animal. In addition, the mutations downstream of the knockout gene well beyond the upstream. This study has found that gene editing can lead to unexpected mutations in the genome, but most of these mutations are harmless. This research has deepened our understanding of CRISPR/Cas9 and broadened its application prospects.

Notoginseng Radix et Rhizoma Powder Treats Dyslipidemia via PI3K/Akt Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo evaluate the clinical efficacy and safety of Notoginseng Radix et Rhizoma powder in treating dyslipidemia by regulating the phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway. MethodSixty patients with dyslipidemia （syndrome of combined phlegm and stasis） treated in the Third Affiliated Hospital of Henan University of Chinese Medicine from May 2021 to June 2022 were selected in this study and randomized into two groups according to the randomized， double-blind control principle. The control group was treated with Xuezhikang capsules + Notoginseng Radix et Rhizoma powder placebo and the observation group with Notoginseng Radix et Rhizoma powder + Xuezhikang capsules placebo for 6 weeks. The clinical efficacy， traditional Chinese Medicine （TCM） syndrome scores， and liver and kidney function indicators were evaluated at weeks 0， 3， and 6. Enzyme-linked immunosorbent assay （ELISA） was employed to determine the expression of vascular endothelial growth factor （VEGF）， kinase insert domain receptor （KDR）， epidermal growth factor （EGF）， and epidermal growth factor receptor （EGFR） in the peripheral serum. Quantitative Real-time PCR was employed to measure the mRNA levels of KDR， EGFR， PI3K， and Akt in the mononuclear cells of the peripheral blood. ResultThe observation group （83.33%） showed the total effective rate comparable to that of the control group （89.66%） and no adverse reactions. Compared with before treatment， the patients in the observation group showed decreased TCM syndrome score and serum levels total cholesterol （TC）， triglyceride （TG）， and low-density lipoprotein cholesterol （LDL-C） and after being treated for 3 and 6 weeks （P<0.05）， the level of high-density lipoprotein cholesterol （HDL-C） showed an upward trend, but the difference was not statistically significant. After treatment， the two groups showed no significant differences. Compared with that before treatment, the mRNA expression of PI3K, Akt and EGFR in peripheral blood mononuclear cell and the expression of EGF, VEGF and KDR in serum of the observation group showed a downward trend with time, in which the mRNA expression of PI3K, Akt, VEGF and KDR decreased more significantly (P<0.05)，The expression levels of KDR mRNA and serum EGFR show a trend of first increasing and then decreasing.Compared with the control group after treatment, there was no statistically significant difference in mRNA expression of PI3K, Akt, EGFR, and KDR, as well as serum levels of EGF, EGFR, VEGF, and KDR between the two groups of patients at the same time point. ConclusionNotoginseng Radix et Rhizoma powder is safe and effective in correcting dyslipidemia. It may inhibit the PI3K/Akt signaling pathway by down-regulating the expression of VEGF/KDR to lower the blood lipid level.

Impact of digitalization on clean governance: An analysis of China's experience of 31 provinces from 2019 to 2021.

The goal of deepening institutional reforms was to bring transparency and accountability, address corruption, and establish a clean government (CG) in China. The first step toward this transparency is considered to be the free development and transmission of Open Data (OD). In this regard, China has set up open data centers in provincial governments. Considering that OD can have an impact on CG and bring new ideas for CG construction, ODs of 31 provincial governments have been analyzed through fsQCA3.0 to test these assumptions. To see how much it can contribute to the development of the Technology Organization Environment Framework (TOE). To this end, between 2019 and 2021, 31 provincial government data have been clustered into low, medium, and high corruption case enrollment areas to determine the impact of OD. The study mentioned that improvements in ODs in 31 provinces could strengthen cooperation with the disciplinary inspection department in the fight against corruption. The study, on the other hand, made two assumptions that environmental barriers and internal pressures could affect data's reliability.

Treatment of vitiligo with fire needle: A protocol for systematic review and meta analysis.

BACKGROUND: We aim to study the treatment of vitiligo with fire needle. METHODS: We will search PubMed, Embase, the Cochrane Library, the China National Knowledge Infrastructure, Chinese Science and Technology Periodical Database, Wanfang Database, and Chinese Biomedical Literature Database of randomized controlled trials beginning from their inception to August 2020. The primary outcomes is that Complex area of white spot and Percentage of Clinical Effectiveness will be accepted as the primary outcomes. Additional outcome is the safety assessment will be considered a secondary outcome. Two independent authors will based on the Cochrane system evaluation manual 5.1.0 version of randomized controlled trial bias risk assessment tool to evaluate the risk of bias among the final included studies. And we will use the RevMan 5.3 software to analysis data. CONCLUSION: This study will provide evidence to judge whether fire needle is an effective therapy for vitiligo. INPLASY REGISTRATION NUMBER: INPLASY2020120012.

Genetic quality analysis of 24 domestic inbred mouse strains by microsatellite DNA / 中国比较医学杂志

Objective To analyze the genetic quality of 24 domestic inbred strains mice using microsatellite loci panel.Methods Previously selected 30 microsatellite loci of mouse with high polymorphism and more allele numbers were used to synthesize corresponding fluorescently-labeled primers.Then the genomic DNA samples of each mouse were amplified by PCR and the products were analyzed by STR scanning to genotype the inbred strains of mice.Results Out of the 24 inbred strains, 15 inbred strains showed the same genotype within one strain at 30 loci.Among different strains, microsatellite loci indicated polymorphism which could be used to distinguish different strains.However, the rest 9 strains demonstrated polymorphism within strains.Conclusions Our stuoly provides a useful microsatellite panel to detect genetic quality of inbred mice and distinguish different strains with the optimized microsatellite loci.

Establishment of a detection method for Helicobacter pylori in living Mongolian gerbil / 中国比较医学杂志

Objective To establish a detection technique for H.pylori(HP) infection in Mongolian gerbils using nested PCR technique.Methods H.pylori was cultured in vitro and inoculated into Mongolian gerbils.At the 10th week after infection, the HP in the gastric juice of Mongolian gerbil was detected by conventional PCR assay and the gastric juice, gastric mucosa, duodenal contents and colon stool were examined by nested PCR.Rapid urease test and ELISA were used to analyze the accuracy of the nested PCR assay.All of the PCR products were verified by sequencing.Results The positive rate of gastric juice detected by conventional PCR was 30%, while the positive rates of gastric juice, gastric mucosa, duodenal contents and colon stool detected by nested PCR were 100%, 100%, 90%, and 10%, respectively.The positive detection rates of rapid urease test and serum ELISA were 100% and 0%, respectively.Comparing the results of different methods, both the positive rates of gastric juice and gastric mucosa detected by nested PCR and the detection rate of rapid urease test were 100%, but the results of conventional PCR detection of gastric juice, the nested PCR detection result of stool in colon and of serum ELISA assay were lower than other methods.Conclusions Due to its high accuracy and sensitivity, the nested PCR assay of gastric juice can be used for the long-time detection of H.pylori infection in Mongolian gerbils, especially useful in the experiments of prevention and treatment of H.pylori infection.

Clinical Observation on Efficacy of Low -calorie Enteral Nutrition Therapy for Postoperative Recovery in Patients with Primary Hepatic Carcinoma / 医学研究杂志

Objective To study the clinical efficacy of low -calorie enteral nutrition support therapy for postoperative recovery in patients with primary hepatic carcinoma .Methods Retrospective analysis was performed in this study .120 patients, including partial liver or local liver resection , were divided into Hypocaloric parenteral nutrition group ( HPN) and Hypocaloric parenteral nutrition group ( HEN) by different nutritional treatment ways with each group of 60 case.Liver function ( TBIL, ALT, AST ) , nutrition indicators (ALB, PCHE, PA), immune function (CD4 +T cells, CD8 +T cells, CD4 +T cells/CD8 +T cell ratio), gastrointestinal tract function ( postoperative intestinal ventilation ) , the incidence of complications and nutritional support costs and hospitalization time difference were compared between two groups .Results Compared with the low -calorie parenteral nutrition group , TBIL, ALT, AST were significantly lower on low calorie enteral nutrition group (P<0.05); ALB, PCHE, PA, ALB, PCHE, PA, CD4 +T cells, CD8 +T cells, CD4 +T cells/CD8 +T cell ratio were significantly higher on low calorie enteral nutrition group ( P<0.05);low-calorie gastrointestinal function recovery, incidence of complications during hospitalization and nutritional support costs on enteral nutrition group were superior to low -calorie parenteral nutrition group .Moreover, the differences were statistical significance (P<0.05).Conclusion For postoperative pa-tients with primary hepatic carcinoma , both enteral and parenteral nutrition support therapies have significant effect , however ,enteral nu-trition therapy has more advantages , better clinical efficacy by comparing to parenteral nutrition therapy .Therefore, it is worthy of apply-ing in clinic .

Conflict Analysis and Countermeasures for Doctor-patient Communication from Both Doctors and Patients′Perspectives / 中国医学伦理学

From both doctors and patients′different perspectives , this paper analyses the causes of conflict of doctor-patient communication , and from the subjective and the objective , the history and reality , individual and social aspects of light on the restricting factors for good doctor -patient communication , and puts forward strengthe-ning the consciousness of active , rule execution , achieving full coverage , explore ways to standard mode , rich di-verse ways and other countermeasures .

The optimization scheme of Mongolian gerbils superovulation induced by PMSG and hCG / 中国比较医学杂志

Objective To optimize the scheme of Mongolian gerbil superovulation.Methods Based on the analysis of the animal-week-old, the dose and time interval of hormone, we got the best animal-week-old, dose and time interval of hormone for Mongolian gerbil superovulation.Results The 6 week old female Mongolian gerbils which injected of 10 IU PMSG and followed by 10 IU hCG in 70 hours later could get the best superovulation.We collected eggs at 16 hour after mate with male gerbils.The ovum pick-up rate reached 80%, the number of oocytes were 32.6 ±3.0, the number of the fertilized egg developed to 2-cell are 24.8 ±5.4.Conclusions This study summarizes the optimization scheme of Mongolian gerbil superovulation induced by PMSG and hCG and it supported the foundation for Mongolian gerbils embryo biotechnology.

Application of self-made quality control materials on chemiluminescence analyzer / 国际检验医学杂志

Objective To investigate the feasibility of self-made quality control materials on chemiluminescence analyzer .Meth-ods Residual serum in blood bags were collected and served as matrixes .Complex quality control materials of tumor markers [car-cinoembryonic antigen ,alpha-fetoprotein ,carbohydrate antigen 125(CA125) ,CA199 ,CA153 ,human chorionic gonadotrophin and total prostate specific antigen ] and hormonal markers (luteinizing hormone ,follicle-stimulating hormone ,estradiol ,testosterone , progesterone and prolactin) were prepared .The monthly average value ,coefficient of variation(CV)% value ,cumulative average value and the number of out-of-control of each marker during the 12 months were calculated .Results All self-made quality control serum which stored and packaged separately showed appearance of clear and transparent after freezing and thawing ,without turbidi-ty and sedimentation .Difference of monthly average value and cumulative average value of each marker had no statistical significance (P>0 .05) .Compared with monthly average value and cumulative average value of each marker ,no obvious drift and trends were found .Monthly indoor imprecision of each hormonal marker were controlled within the allowable range .36 times of out-of-control alarm were monitored during a year ,among them ,25 for system error and 11 for random error .Conclusion Self-made quality con-trol materials has good stability and may find the instability factors of detection system timely to guarantee the reliability of test re-sults of chemiluminescence analyzer .

Population genetic quality analysis of outbred KM mice from Shanghai Seed Center / 中国比较医学杂志

Objective To analyze and evaluate the population genetic quality of outbred KM mice from National Rodent Seed Center ( Shanghai ) .Methods A total of 30 outbred KM mice were randomly chosen.The genetic characteristics of the population were determined by PCR and STR scanning using 30 selected microsatellite loci. Popgen1.32 software was used to process the data.Results Thirty microsatellite loci shared 123 alleles in the KM mouse population.The average effective allele number and the average heterozygosity were 2.3989 and 0.5342, respectively.The average polymorphism information content ( PIC ) was 0.4735.Conclusions The outbred KM mouse population of Shanghai Seed Center has genetic stability and genetic diversity, and is satisfied with the genetic characteristics of closed colony laboratory animal.

Advances in development and application of Mongolian gerbil resource / 中国实验动物学报

Mongolian gerbil is a multifunctional laboratory anima1 resource with Chinese characteristics.It has many advantages for some research fields, and play an important role.The more biological characteristics of Mongolian ger-bils discovered with in-depth research will promote diverse application of this resoure.In this paper, we reviewed the devel-opment of application of Mongolian gerbils in taxonomy, parasitology, microbiology, cerebral ischemia, lipid metabolism, neurological diseases, and cancer researches.

Shenkang pill down-regulates AOPP-induced expression of inflammatory factor MCP-1 via a p38MAPK/NF-κB-dependent mechanism / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of the serum of rats fed with Shenkang pill in regulating monocyte chemoattractant protein 1 (MCP-1) expression induced by advanced oxidation protein products (AOPP) in mouse podocyte clone 5 (MPC5) and explore the underlying mechanism.</p><p><b>METHODS</b>MPC5 cultured in vitro were incubated for different time lengths in the presence of different concentrations of serum of rats medicated with Shenkang pill, and the cell proliferation was assessed using MTT assay. In MPC5 treated with AOPP prior to exposure to the rat serum, the changes in the protein expressions of p38MAPK and IκBα were examined with Western blotting, NF-κB p65 nuclear translocation was analyzed with immunofluorescence assay, and MCP-1 expression in the supernatant was determined using ELISA kits.</p><p><b>RESULTS</b>The medicated rat serum time- and concentration-dependently promoted the proliferation of MPC5, with the optimal serum concentration of 5% and incubation time of 24 h. AOPP significantly increased MCP-1 expression in the cell supernatant in a time-and concentration-dependent manner; pretreatment with SB203580 (a p38 inhibitor) or parthenolide (a NF-κB inhibitor) significantly decreased MCP-1 expression, and treatment with the medicated serum significantly decreased AOPP-induced MCP-1 expression. AOPP concentration-dependently increased the protein expression of P-p38 but decreased that of IκBα. Both the medicated serum and SB203580 increased IκBα protein in AOPP-induced cells, but the effect was more obvious with the medicated serum. The medicated serum also decreased NF-κB p65 nuclear translocation in AOPP-induced MPC5.</p><p><b>CONCLUSION</b>Shenkang pill-medicated serum can decrease AOPP-induced expression of MPC-1 in MPC5 by regulating p38MAPK/NF-κB to mediate its anti-inflammatory effect. This finding provides a new theoretical basis for the application of Shenkang pill to treat diabetic nephropathy.</p>

Bone marrow mononuclear cells combined with nano-hydroxyapatite/collagen for repair of mandibular defects / 中国组织工程研究

BACKGROUND:Nano-hydroxyapatite/col agen combined with mononuclear cel s can promote the growth of a variety of stem cel s to induce the formation of new bone and vascularization, final y inducing osteogenesis. OBJECTIVE:To investigate the feasibility of bone marrow mononuclear cel s combined with the nano-hydroxyapatite/col agen for repair of mandibular defects in a rabbit. METHODS:Twenty-seven New Zealand white rabbits were selected to prepare bilateral mandibular bone defect models, and then divided into three groups. In experiment group, bone marrow mononuclear cel s combined with the nano-hydroxyapatite/col agen were implanted into mandibular defects;in control group, nano-hydroxyapatite/col agen scaffold was implanted;and in blank control group, nothing was implanted. Tissue specimens were prepared at weeks 4, 8, 12 for gross observation, imaging analysis, hematoxylin-eosin staining and scanning electron microscope. RESULTS AND CONCLUSION:The imaging examination and histological staining showed that the bone quality and healing degree in the experimental group was better than those in the other groups. Scanning electron microscope showed that better histocompatibility and no inflammation reaction in the experimental group. Dental CT data showed that the experimental group had better repair effect than the other groups (P<0.05). These findings indicate that bone marrow mononuclear cel s combined with the nano-hydroxyapatite/col agen have capacity of bone induction and bone formation, which can be used to repair mandibular defects.

The expression of tumor necrosis factor-α and its significance in different gestational period placentas and aging placentas / 中国基层医药

ObjectiveTo study the differences of tumor necrosis factor-α in different gestational period placentas and aging placentas and to analyze its significances. MethodsUsing immunohistochemistry to detect the expression and distribution of tumor necrosis factor-α in different gestational period placentas and aging placentas. ResultsThe expression of tumor necrosis factor-α was significantly higher in the third trimester of pregnancy placentas and aging group than in the first trimester of pregnancy placentas(P＜0.05),and than in the second trimester of pregnancy placentas(P＜0.05).The expression of tumor necrosis factor-α had no significant difference between the third trimester of pregnancy placentas and aging group(P＞0.05). ConclusionTNF-α should play an important role in placental aging.

Ultrasound-guided minimally invasive stereotactic breast excision for breast mass / 中国医师进修杂志

Objective To evaluate the application value of ultrasound-guided minimally invasive stereotactic breast excision in the diagnosis and treatment of breast mass. Methods The minimally invasive operations for 218 patients were performed by ultrasound-guided Mammotome system, and the therapeutic efficacy of Mammotome system was evaluated. Results All the patients were completely excised by the Mammotome system. No failed procedure occurred,all the masses were pathologically confirmed as benign lesions in 214 patients and invasive ductal carcinomas in 4 patients. Postoperatively, the incision scar was barely visible and no complications happened except for mild subcutaneous hemorrhage in 9 patients,local hematoma in 3 patients and local deep pitting of the skin in 5 patients. Conclusion Mammotome system shows many benefits with less trauma, more rapid recovery and with less complications.

Comparison between three-dimensional cephalometric and three-dimensional CT measurements of adult zygomatic complex at Sanjiang area / 中华医学美学美容杂志

Objective To compare the three-dimensional eephalometric and three-dimensional CT measurements of adult zygomatic complex among a Sanjiang population.Methods 120 female natives of Sanjiang region from Chinese Han between 19 to 23 years of age (mean 21.4 years),were randomly selected from students of Jiamusi University.Three-dimensional cephalometric and CT measurements of the face and skull were taken,with reference to zygomatic size measurement by Prof.Qi Zuo-liang,for the comparative study, which included the upper,mid and lower facial width followed by the length,width and angle of malar process,and width of the facial bone.Statistical analysis was done with the obtained measurements.Results Three-dimensional CT analysis showed facial width ratio of 0.83 and 0.79 and skeletal facial profile width ratio of 0.81 and 0.77,respectively,when compared with three-dimensional cephalometric analysis that showed facial width ratio of 0.84 and 0.77.Both values showed no statistical significance (P>0.05).Conclusion Three-dimensional CT measurement as the same to three-dimensional cephalometric can be used in the diagnosis of prominent malar complex.

Expression of tissue factor in the bone marrow of multiple myeloma patients and its relationship with angiogenesis / 白血病·淋巴瘤

Objective To detect the expression of tissue factor(TF) in bone marrow (BM) of patients with multiple myeloma (MM), and to analyze the relationship among TF, vascular endothelial growth factor (VEGF) and the angiogenesis in MM. Methods TF, VEGF and mierovessel density (MVD) levels were detected by immunohistochemistry technique for bone marrow biopsy from 32 cases of newly diagnosed or relapsed MM patients and 32 cases of iron deficiency anemia or healthy volunteers. Results In bone marrow TF and VEGF positive rate in MM group and control group is 88 % and 78 %; 60 % and 10 % respectively. Median TF and VEGF expression levels in MM group and control group were ++ and ++; - and - respectively, median MVDs were 4and O. Conclusion Both 'IF and VEGF expression in MM bone marrow are higher than those in normal bone marrow; and correlates with angiogenesis positively.

Removal of artifacts from EEG signal / 生物医学工程学杂志

As a kind of physiological signals, the electroencephalogram (EEG) represents the electrical activity of the brain. Because of its higher time-varying sensitivity, EEG is susceptible to many artifacts, such as eye-movements, blinks, cardiac signals, muscle noise. These noises in recording EEG pose a major embarrassment for EEG interpretation and disposal. A number of methods have been proposed to overcome this problem, ranging from the rejection of various artifacts to the effect estimate of removing artifacts. This paper reviews many kinds of methods for artifact rejection in the EEC recently, including regression-based methods, artifact subtraction, principal component analysis (PCA), independent component analysis (ICA) and wavelet transform. The specific assumptions of each method and its advantage/disadvantage are also summarized.

Relationship between mutations of the hyper-variable region of mitochondrial DNA D-loop and clinical characteristics in laryngeal cancer / 中国耳鼻咽喉头颈外科

OBJECTIVE To investigate the features of somatic mutations of the hypervariable region (HV) of mitochondrial DNA (mtDNA) D-loop in laryngeal squamous cell carcinoma (LSCC) and the relationship with clinicopathological characteristics. METHODS Microdissected cancerous tissues, paracancerous tissues and peripheral vein blood were obtained from 40 patients with LSCC and total DNA was extracted. HVⅠ and HVⅡ fragments were amplified by PCR and purified. Mutations were determined by DNA sequencing. The mutations of HVⅠand HVⅡ and their relationship with clinicopathological characteristics were analyzed. RESULTS ①In 40 LSCC samples, 86 polymorphisms were found, 21 (52.5 %) tumors presented somatic mtDNA mutations with a total of 34 mutations. Among them, 28 (82.4 %) and 6 (17.6 %) were located in HVⅠ and HVⅡ respectively. ②Tumor D-loop mutation showed significant correlation with tumor differentiation(P=0.028) and p53 mutation (P=0.041), respectively. CONCLUSION Mitochondrial DNA D-loop in LSCC is an unstable region with high frequency of somatic mutation and high polymorphism and might play a potential role in the carcinogenesis of the larynx.