Objective To study the influence of diabetes on the CK activity in different tissues of streptozotocin induced diabetic rats.Methods Serum samples,heart and skeletal muscles,brain and bladder tissues were collected from both streptozocin induced diabetic rats and control group.CK was measured by enzymatic method.Results The body weight,heart weight,and brain weight reduced significantly compared with control group(P

Objective To analyze the clinical distribution ,antimicbial resistance and Staphylococcal cassette chromosome mec (SCCmec) genotype characteristics of 346 methicillirrresistant Staphylococcus aureus (MRSA) clinical isolates in the hospital . Methods A total of 784 strains of Staphylococcus aureus isolated from January 2014 to January 2015 in the hospital ,MRSA identi-fication and the SCCmec genotype was conducted by PCR assay .Results 346 strains of MRSA (44 .13% ) were isolated from 784 strains of Staphylococcus aureus ,the detection rate of MRSA from sputum accounted for 43 .06% ,the secretion accounted for 48 .55% .MRSA was resistant to penicillin ,levofloxacin and erythromycin ,sensitive to vancomycin ,linezolid and teicoplanin .SCC-mec genotyping result showed that SCCmecⅡ was identified in 130 ,SCCmecⅢ in 196 ,SCCmecⅣ in 11 ,SCCmecⅤ in 9 .Conclusion SCCmecⅢ is the main genotypes of MRSA from our Hospital ,all of the MRSA strains are multi-resistant to tested antibiotics , but sensitive to vancomycin and teicoplanin .

OBJECTIVE To study the mutation profiles of hepatitis B virus(HBV) in the core regions.METHODS Based on the sequence alignment of all HBV genotypes,specific primers targeting all HBV genotypes were designed to amplify the core region of HBV followed by sequence analysis on the sequencing data available.RESULTS Among the 34 cases,23 cases showed mutations in the core region.According to the mutation profiles,the most common mutations were the A1762T(50.0%) and G1764A(52.9%) in the basic core promoter(BCP) regions,and it always showed as double mutations.The L60V in core gene regions was the secondary common mutations(17.6%).Among all patients,there were 18,6 and 10 cases showed mutations in BCP,pre-core,and core gene regions,respectively.The most common mutations in BCP,pre-core,and core gene regions were the double mutations at A1762T and G1764A(94.7%),G1896A(83.3%),and L60V(50.0%),respectively.CONCLUSIONS The most common mutations in the core region of HBV are the double mutations at A1762T and G1764A.Analysis on the mutation profiles of HBV core regions might be helpful for the prognosis and prediction of HBV infections.

Objective:To analyze the composition differences of intestinal microbiota in patients with colon cancer and rectal cancer.Methods:The fecal samples of 72 patients in the Second Affiliated Hospital of Harbin Medical University from July 2018 to January 2019 were collected, and they were divided into colon cancer group and rectal cancer group, 36 cases in each group. DNA from fecal samples was extracted, and then high-throughput sequencing was performed on DNA. Bioinformatics was used to analyze the diversity and composition differences of intestinal microbiota between the two groups, and the potential cancer-promoting mechanisms of the differential flora were also discussed.Results:From high-throughput sequencing, 2 356 560 original sequences and 32 730 high-quality sequences were obtained from 72 samples. The average length of the sample sequence was mainly in the interval of 401-460 bp. And 1 409 operational taxonomic units (OTU) were acquired after OTU species taxonomy annotation of all the sequences. Alpha diversity analysis showed that Shannon index of the rectal cancer group and the colon cancer group was 2.61±0.56 and 2.43±0.67, respectively, and the difference was statistically significant ( t = 1.229, P = 0.223); Simpson index of the rectal cancer group and the colon cancer group was 0.17±0.09 and 0.21±0.16, respectively, and the difference was statistically significant ( t = 1.449, P = 0.151). Differences analysis of both groups and linear discriminant analysis (LDA) showed at the phylum level, Firmicutes were more abundant in the intestine of patients with rectal cancer (LDA = 4.67, P = 0.014), while Proteobacteria were more abundant in the gut of colon cancer patients (LDA = 4.49, P = 0.042). From the perspective of class level, the abundance of Gammaproteobacteria was higher in the intestine of patients with colon cancer (LDA = 4.50, P = 0.033), while the abundance of Erysipelotrichia was higher in the intestine of patients with rectal cancer (LDA = 3.50, P = 0.035). At the order level, the abundance of Erysipelotrichales was higher in the intestine of patients with rectal cancer (LDA = 3.50, P = 0.035); at the family level, the abundance of Porphyromonadaceae was higher in the intestine of patients with rectal cancer (LDA = 3.97, P = 0.033). Conclusion:The compositions of intestinal microbiota in patients with colon cancer and rectal cancer are significantly different, indicating that the different floras may contribute to the progression of colon cancer and rectal cancer.

Aim To evaluate the antitumor activity of dianhydrogalactitol ( DAG) in vitro, and further clarify its underlying mechanisms. Methods The inhibitory effect of DAG in NCI-H460 cells was detected by CCK-8 assay and colony formation assay. The morphology of cells treated with DAG was observed under optical mi-croscope. Nuclear morphology was captured by fluores-cence microscopy after Hoechst 33342 staining. Real-time PCR was used to analyze the expression level of topoisomerase Ⅱ ( Topo Ⅱ) mRNA. The protein ex-pression level of Topo Ⅱ was detected by Western blot. Additionally, molecular docking approaches were used to predict the interaction between DAG and TopoⅡ. Results DAG exhibited potent antitumor activity in NCI-H460 cells, and inhibited cell proliferation per-sistently. DAG obviously induced nuclear morphologi-cal changes of NCI-H460 cells. Furthermore, DAG could down-regulate the mRNA and protein expression level of Topo Ⅱ detected by Real-time PCR analysis and Western blot, respectively. Molecular docking predicted that DAG could bind to Topo Ⅱ. Conclu-sion DAG can significantly inhibit the proliferation of NCI-H460 cells, and its underlying mechanisms may involve the down-regulation of Topo Ⅱ mRNA and di-rect binding to Topo Ⅱ, leading to cancer cell death.

OBJECTIVE: To analyze the genomic variation characteristics of fetal with abnormal serological screening, and to further explore the value of copy number variation (CNV) detection technology in prenatal diagnosis of fetal with abnormal serological screening. METHODS: 7617 singleton pregnant women who underwent amniocentesis for prenatal diagnosis solely due to abnormal Down's serological screening were selected. According to the results of serological screening, the patients were divided into high risk group, borderline risk group and single abnormal multiple of median (MOM) group. CMA and CNV-Seq were used to detect the copy number variation of amniotic fluid cell genomic DNA and combined with amniotic fluid cell karyotype analysis for prenatal diagnosis. Outpatient revisit combined with telephone inquiry was used for postnatal follow-up. RESULTS: Among 7617 amniotic fluid samples, aneuploidy was detected in 138cases (1.81%) by CMA and CNV-Seq, 9 cases of aneuploid chimerism were detected by amniotic fluid cell karyotype analysis, and 203 cases of fetus carrying pathogenic and likely pathogenic CNV (P/LP CNV) were detected, the variant of uncertain significance (VUS) was detected in 437 cases (5.7%), the overall abnormal detection rate was 10.33%. The detection rate of aneuploidy by CMA and CNV-Seq in three group were 123 cases (2.9%), 13 cases (1.3%) and 2 cases (0.4%), respectively,and showing no significant difference (χ ²=7.469, P=0.024). The detection rate of pathogenic and likely pathogenic CNV in three group were 163cases (2.6%); 24 cases (2.6%) and 16 cases (3.3%), respectively, and showing no significant difference (χ ²=0.764, P=0.682). The CMA reported 2.9% (108/3729)P/LP CNV, and CNV-seq reported 2.4% (95/3888)P/LP CNV, both tests showed similar detective capabilities (χ ²=1.504, P=0.22).The most popular P/LP CNV in this cohort were Xp22.31 microdeletion, 16p13.11 microduplication /microdeletion, 22q11.21 microduplication /microdeletion. In fetuses with P/LP CNV CNV, 59 fetuses were terminated pregnancy, and 32 of 112 fetuses born had abnormal clinical manifestations. Non-medically necessary termination of pregnancy occurred in 11 fetuses carrying VUS CNV, 322 fetuses carrying VUS CNV were born, 4 of them presented abnormal clinical manifestations. CONCLUSION Compared with the traditional chromosome karyotype, CMA and CNV-Seq can improve the detection rate of pathogenic and likely pathogenic CNV. CMA and CNV-seq can be used for first tier diagnosis of pregnant women in the general population with abnormal Down's serological screening.
Amniotic Fluid ; Aneuploidy ; Chromosome Aberrations ; DNA Copy Number Variations ; Female ; Genomics ; Humans ; Pregnancy ; Pregnancy Trimester, Second ; Pregnant Women ; Prenatal Diagnosis/methods* ; Technology

Objective:To analyze the genetic etiology and prognosis in fetuses with increased nuchal translucency (NT) in order to assist in the clinical prenatal genetic counseling and diagnosis.Methods:This study retrospectively enrolled 1 658 cases of singleton pregnancy (<35 years old) receiving invasive prenatal diagnosis, including karyotype analysis and/or chromosome microarray analysis or copy number variation (CNV) sequencing, due to NT value ≥2.5 mm in the first trimester in Henan Provincial People's Hospital from August 2014 to December 2021. They were divided into different groups according to the thickness of NT (≥2.5-<3.0, ≥3.0-<3.5, ≥3.5-<4.5, ≥4.5-<5.5, ≥5.5-<6.5 and ≥6.5 mm groups) and abnormal ultrasound findings (isolated increased NT group, increased NT complicated by soft markers/non-severe structural abnormality group and increased NT complicated by severe structural abnormality group). The results of invasive prenatal diagnosis and pregnancy outcomes were compared between different groups using Chi-square test and trend Chi-square test. Results:The detection rates of numerical abnormalities of chromosomes were 15.8% (262/1 658) and 17.6% (252/1 431) when the NT thickness cut-off value were 2.5 mm or 3.0 mm, respectively. Overall, the detection rate of numerical abnormalities of chromosomes increased with thickness of NT ( χ2trend=180.75, P<0.001), ranging from 6.6% (44/671) in the NT≥2.5-<3.5 mm group to 45.6% (113/248) in the NT≥5.5 mm group. The incidence of pathogenic/likely pathogenic CNV(P/LP CNV) did not increased with NT thickness ( χ2trend=3.26, P=0.071), and the highest detection rate was observed in the NT≥4.5-<5.5 mm group (9.0%, 19/211). The detection rate of numerical abnormalities of chromosomes plus P/LP CNV in the isolated NT≥2.5-<3.0 mm group and NT≥3.0-<3.5 mm group were 5.3% (10/188) and 9.6% (36/375), respectively, however, the difference was not statistically significant ( χ2=3.06, P=0.080). The detection rates of numerical abnormalities of chromosomes plus P/LP CNV in the isolated NT≥3.5-<4.5 mm group and NT≥2.5-<3.0 mm complicated by soft markers/ non-severe structural abnormality group were 12.7% (52/410) and 24.1% (7/29), respectively, and the risk were 2.6 times (95% CI: 1.3-5.2) and 5.7 times (95% CI: 2.0-16.4) of the isolated NT≥2.5-<3.0 mm group, respectively. The pregnancy termination rate increased with the NT thickness ( χ2trend=304.42, P<0.001), ranging from 10.8% (23/212) in the NT≥2.5-<3.0 mm group to 90.7% (117/129) in the NT≥6.5 mm group. After exclusion of the pregnancies terminated due to numerical abnormalities of chromosomes and P/LP CNV, 87.6% (862/984) of the fetus with increased NT were born alive. Conclusions:The detection rate of numerical abnormalities of chromosomes increases with the thickness of NT. Invasive prenatal diagnosis is required for non-advance aged singleton pregnant women when fetuses present with isolated NT≥2.5 mm with or without soft markers/structural abnormalities.

Objective:To investigate the clinical efficacy of radical resection of rectal cancer with different surgical approaches and influencing factors of postoperative complications.Methods:The retrospective study was conducted. The clinicopathological data of 3 418 patients who underwent radical resection of rectal cancer in the Second Affiliated Hospital of Harbin Medical University from July 2011 to September 2020 were collected. There were 2 060 males and 1 358 females, aged (61±11)years. Patients meeting the requirements of radical resection and surgical indications underwent surgeries choosing from open radical colorectal cancer surgery, laparoscopic radical colorectal cancer surgery, and natural orifice specimen extraction surgery (NOSES). Observation indicators: (1) intraoperative and postoperative conditions of patients undergoing different surgical approaches; (2) comparison of preoperative clinical characteristics in patients undergoing different surgical approaches; (3) comparison of postoperative histopathological characteristics in patients undergoing different surgical approaches; (4) postoperative complications of patients undergoing different surgical approaches; (5) analysis of influencing factors of postoperative complications. Measurement data with normal distribution were represented as Mean± SD. Measurement data with skewed distribution were represented as M(range), and comparisons between groups was analyzed using the Kruskal-Wallis rank test. Comparison of ordinal data was analyzed using the non-parameter rank sum test. Count data were described as absolute numbers or percentages, and comparison between groups was analyzed using the chi-square test. Multivariate analysis was conducted using the Logistic regression model. Results:(1) Intraoperative and postoperative conditions of patients undergoing different surgical approaches. Of the 3 418 patients, 1 978 cases underwent open radical colorectal cancer sur-gery, 1 028 cases underwent laparoscopic radical colorectal cancer surgery and 412 cases underwent NOSES, respectively. The operation time, volume of intraoperative blood loss, cases with permanent stoma, preventive stoma or without fistula, time to postoperative first flatus, time to postoperative liquid food intake, cases transferred to intensive care unit after surgery, duration of postoperative hospital stay were 145(range, 55?460)minutes, 100(range, 30?1 000)mL, 435, 88, 1 455, 72(range, 10?220)hours, 96(range, 16?296)hours, 158, 10(range, 6?60)days, respectively, in patients undergoing open radical colorectal cancer surgery. The above indicators were 175(range, 80?450)minutes, 50(range, 10?800)mL, 172, 112, 744, 48(range, 14?120)hours, 72(range, 38?140)hours, 17, 9(range, 4?40)days, respectively, in patients undergoing laparoscopic radical colorectal cancer surgery and 180(range, 80?400)minutes, 30(range, 5?500)mL, 0, 45, 367, 48 (range, 14?144)hours, 72(range, 15?148)hours, 1, 6(range, 3?30)days, respectively, in patients undergoing NOSES. There were significant differences in the above indicators among the patients undergoing different surgical approaches ( H=291.38, 518.56, χ2=153.82, H=408.86, 282.97, χ2=78.66, H=332.30, P<0.05). (2) Com-parison of preoperative clinical characteristics in patients undergoing different surgical approaches. The gender, age, body mass index, cases with diabetes, cases with hypertension, cases with coronary heart disease, cases with anemia, cases with hypoproteinemia, cases with intestinal obstruction, tumor location, preoperative carcinoembryonic antigen, preoperative CA19-9 showed significant differences among patients undergoing open radical colorectal cancer surgery, laparoscopic radical colorectal cancer surgery and NOSES ( P<0.05). (3) Comparison of postoperative histopathological characteris-tics in patients undergoing different surgical approaches. Tumor histological type, tumor differentiation degree, tumor diameter, number of lymph node detected, nerve invasion, vascular invasion, lymph node invasion, tumor T staging, tumor N staging, tumor M staging, tumor TNM staging showed significant differences among patients undergoing open radical colorectal cancer surgery, laparos-copic radical colorectal cancer surgery and NOSES ( P<0.05). (4) Postoperative complications of patients undergoing different surgical approaches. Cases with postoperative complications as anastomotic leakage, abdominal infection, intestinal obstruction, anastomotic bleeding, incision complications, pulmonary infection, other complications were 52, 21, 309, 8, 130, 51, 59, respectively, in patients undergoing open radical colorectal cancer surgery. The above indicators were 33, 17, 75, 3, 45, 58, 9, respectively, in patients undergoing laparoscopic radical colorectal cancer surgery and 13, 4, 8, 0, 11, 10, 15, respectively, in patients undergoing NOSES. There were significant differences in the intes-tinal obstruction, incision complications, pulmonary infection, other complications among patients undergoing different surgical approaches ( χ2=122.56, 13.33, 20.44, 15.59, P<0.05) and there was no significant difference in the anastomotic leakage, abdominal infection, anastomotic bleeding among patients undergoing different surgical approaches ( χ2=0.96, 2.21, 3.08, P>0.05). (5) Analysis of influencing factors of postoperative complications. ① Analysis of influencing factors of intestinal obstruction in patients with radical resection of rectal cancer. Age as 20?39 years and 40?59 years, surgical approach as laparoscopic radical colorectal cancer surgery and NOSES were independent protective factors of intestinal obstruction in patients with radical resection of rectal cancer ( odds ratio=0.46, 0.59, 0.43, 0.13, 95% confidence interval as 0.21?1.00, 0.36?0.96, 0.33?0.56, 0.06?0.27, P<0.05). ② Analysis of influencing factors of incision complications in patients with radical resection of rectal cancer. Body mass index as 24.0?26.9 kg/m 2, surgical approach as laparoscopic radical colorectal cancer surgery and NOSES were independent protective factors of incision complications in patients with radical resection of rectal cancer ( odds ratio=0.24, 0.63, 0.46, 95% confidence interval as 0.11?0.51, 0.44?0.89, 0.24?0.87, P<0.05). ③ Analysis of influencing factors of pulmonary infection in patients with radical resection of rectal cancer. The surgical approach as laparoscopic radical colorectal cancer surgery was an independent risk factor of pulmonary infection in patients with radical resection of rectal cancer ( odds ratio=2.15, 95% confidence interval as 1.46?3.18, P<0.05), and tumor TNM staging as 0?Ⅰ stage was an independent protective factor ( odds ratio=0.10, 95% confidence interval as 0.01?0.88, P<0.05). ④ Analysis of influencing factors of other complica-tions in patients with radical resection of rectal cancer. Age as 20?39 years, 40?59 years, 60?79 years, body mass index as <18.5 kg/m 2, 18.5?23.9 kg/m 2, 24.0?26.9 kg/m 2, 27.0?29.9 kg/m 2, surgical approach as laparoscopic radical colorectal cancer surgery were independent protective factors of other complications in patients with radical resection of rectal cancer ( odds ratio=0.10, 0.29, 0.37, 0.08, 0.22, 0.35, 0.32, 0.29, 95% confidence interval as 0.01?0.81, 0.13?0.64, 0.17?0.78, 0.02?0.40, 0.09?0.52, 0.15?0.83, 0.12?0.89, 0.14?0.59, P<0.05). Conclusions:Compared to laparoscopic radical colorectal cancer surgery and NOSES, open radical colorectal cancer surgery has wide indication and short operation time, but less perioperative treatment effect. Laparoscopic radical colorectal cancer surgery and NOSES can achieve better surgical result and less postoperative complication when patients meeting surgical indications.

Objective:To analyze the characteristic of prenatal serological screening in fetus with X-linked ichthyosis (XLI), and to explore the relationship between unconjugated estriol (uE 3) levels and XLI. Methods:A total of 56 fetuses with Xp22.31 microdeletion indicated by prenatal diagnosis and 70 fetuses diagnosed with trisomy 21 and 26 fetuses with trisomy 18 in Henan Provincial People's Hospital and Affiliated Hospital of Weifang Medical College from September 2016 to June 2021 were collected. The multiples of median (MoM) values of uE 3, alpha-fetoprotein (AFP), and human chorionic gonadotropin (hCG) during the second trimester of pregnancy were retrospectively analyzed. Prenatal diagnosis was made by amniotic fluid karyotype analysis and genome copy number variant analysis, parent genetic verification and pathogenicity analysis were performed, and maternal and infant outcomes were followed up. Results:Of 56 pregnant women with fetal Xp22.31 microdeletion, 43 underwent serological screening during the second trimester of pregnancy, of which 42 were abnormal (39 male fetuses and 3 female fetuses). The median uE 3 MoM value of 39 male fetuses [0.06 (0.00-0.21)] was lower than the normal value and significantly lower than that of fetuses with trisomy 21 [0.71 (0.26-1.27)] and fetuses with trisomy 18 [0.36 (0.15-0.84)], the difference was statistically significant ( Z=99.96, P<0.001). While the MoM values of AFP and hCG were all within the normal range. Among the 56 fetuses carrying Xp22.31 microdeletion, 45 were male fetuses and 11 were female fetuses, and the deletion fragments all involved STS gene. Eighty-nine percent (50/56) were inherited from mother (49 cases) or father (1 case), and 11% (6/56) were de novo mutations. Follow-up showed 48 live births (38 males and 10 females) and 8 chose to terminate pregnancy (7 males and 1 female). Among the 38 male newborns, 37 presented with scaly skin changes from 1 to 3 months of age, and one had no clinical manifestations until 4 months after birth. Ten female newborns had no obvious clinical manifestations. Conclusions:The decrease levels of uE 3 MoM on maternal serological screening is closely related to the higher risk of XLI in male fetuses. For pregnant women with low uE 3 in serological screening or with family history of ichthyosis, in addition to chromosomal karyotype analysis, joint detection of genomic copy number variant analysis should be recommended.

OBJECTIVE：To compare the effects of raw pro duct and different processed products of Gekko gecko on kidney-yang deficiency model mice induced by adenine. METHODS ：Totally 100 mice were randomly divided into blank group （n＝10）and modeling group （n＝90）. Modeling group was given adenine （50 mg/kg）intragastrically for 10 days to induce kidney-yang deficiency model ；blank group was given normal saline （0.2 mL/10 g）intragastrically. After modeling ，70 mice were randomly divided into model group ，positive group （Jinkui shenqi pill ，6.4 g/kg），G. gecko crude product group （1.2 g/kg）， wine-processed G. gecko group（1.2 g/kg）and oil-processed G. gecko group（1.2 g/kg）according to body weight and symptoms of kidney-yang deficiency ，with 14 mice in each group. Blank group and model group were given constant volume of normal saline intragastrically；administration groups were given relevant medicine intragastrically （0.2 mL/10 g），once a day ，for consecutive 14 d. During the experiment ，the symptoms and signs of mice in each group were observed. After last medication ，renal index ，testis index and serum levels of T ，CORT，BUN and Cr were measured ；HE staining method was used to observe the pathological changes of renal tissue of mice in each group. RESULTS ：Compared with blank group ，the mice in the model group suffered from performance of kidney-yang deficiency ，such as weight loss ，crouch and arch back ，chills and cold limbs ，and sparse body hair ， while renal index and serum levels of BUN and Cr were increased significantly （P＜0.01）. In renal tissue ，there were BA28117 pathological damages such as irregular arrangement of renal KY2016YB211tubular epithelial cells ，light staining of nucleus and edema of cytoplasm. Compared with model group ， performance of kidney-yang deficiency was improved to different extents in G. gecko crude product group and processed product groups（especially in wine-processed G. gecko group）；serum levelsof BUN and Cr were decreased significantly （P＜0.05 or P＜0.01）；pathological damage of renal tissue was alleviated in different degrees. In addition ，body weight of mice was increased significantly in G. gecko processed products groups （P＜0.01），and renal indexes of mice were decreased significantly in G. gecko crude product group and processed products groups （P＜0.05 or P＜0.01）. Compared with G. gecko crude product group ，renal index ，serum levels of BUN and Cr were significantly decreased in wine-processed G. gecko group（P＜0.01），and serum level of Cr was significantly decreased in oil-processed G. gecko group（P＜ 0.05）. CONCLUSIONS ：G. gecko crude product ，wine-processed G. gecko and oil-processed G. gecko all show a certain improvement effect on kidney-yang deficiency mice induced by adenine ，especially wine-processed G. gecko .