Laboratory diagnosis is the main contents of diagnostics course for clinical majors,whose contents is too broad and too difficulty to understand.For this reason,the results of traditional teaching method are not satisfactory.Therefore,teaching and research section did an exploratory reform on experiment diagnosis teaching by clearly defining teaching contents,emphasizing clinical thinking way,reforming experimental teaching,adding multimedia to assist teaching and cultivating high quality teaching teams.As a result,the reform enhanced study interest of students,promoted teaching quality and effects greatly and achieved preliminary success.

OBJECTIVETo establish a model of hypoxic postconditioning of dermal microvascular endothelial cells.

METHODSDuring reoxygenation after hypoxia, cells received three times of hypoxic/ reoxygenation alternate treatment for a certain time. The cells were seeded on 6-well plates, with one plate for one group. They were divided into 6 groups as group 1 ( Control), group 2 (8h hypoxia + 24h reoxygenation), group 3 (8h hypoxia + 2 min x 3 times post-hypoxia treatment) , group 4 (8h hypoxia + 5 min x 3 times post-hypoxia treatment), group 5 (8h hypoxia + 10 min x 3 times post-hypoxia treatment), 6 group (8h hypoxia + 20 min x 3 times post-hypoxia treatment). Each group underwent 8 h hypoxia + 24 h hypoxia Buffer and reoxygenation. Lactate dehydrogenase (LDH) was detected during the process. Apoptosis rate was calculated by staining Tunel method. Bcl-2, Bax and activated caspase-3 protein were detected by Western Blot.

RESULTSIn the continuous hypoxia process, the LDH was (1563 ± 83.35) IU/L at 8h and (582.85 ± 58.25 ) IU/L at 0h, showing a statistical difference (P = 0.0001). Western blotting results showed that the expression of Bax/Bcl-2 in group 2 was 0.38 ± 0.02, showing a significant difference when compared with that in group 3 (0.23 ± 0.01) and group 4 (0.22 ± 0.02) (P = 0.012, P = 0.005), while not when compared with that in group 5 (0.33 ± 0.02) and 6 groups (0.34 ± 0.01) P > 0.05). The ratio of Activated caspase-3/caspase-3 in group 2 (6.30 ± 1.50) was significantly higher than that in group 3 (2.17 ± 0.26) and group 4 (2.63 ± 0.31) (P = 0.008, P = 0.019); while not in group 5 (4.36 ± 0.29) and group 6 (4.97 ± 0.51) (P > 0.05).

CONCLUSIONSThe model of hypoxic postconditioning of human dermal microvascular endothelial cells is successfully established.

Apoptosis ; Blotting, Western ; Caspase 3 ; analysis ; Cell Hypoxia ; Endothelial Cells ; metabolism ; Humans ; In Situ Nick-End Labeling ; Ischemic Postconditioning ; methods ; L-Lactate Dehydrogenase ; analysis ; Oxygen Consumption ; Proto-Oncogene Proteins c-bcl-2 ; analysis ; Time Factors ; bcl-2-Associated X Protein ; analysis

Objective To investigate the clinical effect of Ultrasound-guided minimal invasion technique used in the diagnosis and treatment of benign breast tumor .Methods 60 cases of benign breast tumors , including single or multiple breast lumps ,were treated using minimally invasive technology under the guidance of ultrasound . The average operation time was 25min.Through 3-6months after operations ,any residue and recurrence at local lesion were not found by clinical and ultrasonic examination , and incision scar formation was not clear .Minimally invasive operation can completely amputate breast tumors .Results Under real time ultrasound monitoring ,all the 60 cases of operation had 100%of display rate.The tumor pathologic classification is as follows:25cases of fibroadenomas ,2 ca-ses of cyst ,3 cases breast cystic hyperplasia ,12 cases of adenosis of mammary glands ,3 cases of lipoma of the breast and one case of phyllodes tumor .The other 14 cases have both two kinds of pathological types .Conclusion The system of ultrasound-guided minimally invasive technology can accurately puncture benign breast tumor with high re -section rate,small trauma and is operated safely .It is a therapeutic method and consistent with the opinion of cosme-tology.

Objective To separate and cultivate homo umbilical cord blood (UCB) hematopoietic stem cell (HSC) in vitro, and u-tilize bone marrow desmohemoblast stem cell as trophoblastic layer combined with cytokine to amplify umbilical cord blood hematopoietic stem/progenitor cell. Methods Ficoll lymph-cell separating medium density gradient centrifugalization was used to segregate UCBHSC.Bone marrow desmohemoblast stem ceil and cytokine were added, and the sum of NC cells and CD34 + cells was counted. Results The sum of NC cells amplified 75.2±15.0 times, and the sum of CD 34 + cells amplified 18.7±12.3 times. Conclusions It has significant effect on amplification of hematopoietic stem cell with bone marrow desmohemoblast stem cell and eytokine when HSC are cultured in vitro.

Objective To observe the influence of inactive FRMD4A gene′s expression on the biological behavior of tongue cancer CAL-27cell. Methods FRMD4A-siRNA was transfered into CAL-27 cell by lipidosome, to the expression of FRMD4A-siRNA in CAL-27 cell after transfection was detect by qRT-PCR cell proliferation , was checked by CCK-8,the influence of inactive FRMD4A gene on cell cycle distribution of CAL-27 cell was assayed by flow cytometry. Results Compared with the control group, FRMD4A mRNA expression significantly reduced in FRMD4A-siRNA interfering group (94%) and the cell proliferation index decreased(P<0.05). The cell cycle arrested in G1 period (P<0.05). Conclusion FRMD4A-siRNA could effectively inhibit FRMD4A mRNA expression in tongue cancer CAL-27cell, impact the distribution of cell cycle, and reduce cell proliferation.

Objective To investigate the possible effect of citrate on electrolyte metabolism in healthy people with different genders and races and provide a reference for the possible clinical interventions.Methods A cross over,placebo-controlled study was conducted in 22 age-matched Chinese(11 males and 11 females)and 10 male Caucasian volunteers after informed consents were obtained.Volunteers received of saline solution,separated by a wash-out period of two to three weeks.Serial blood and urine samples were collected during the observation period and analyzed for the selective biochemical parameters.Results Comparable basal levels of serum albumin[male(43.05±1.81)g/L vs female(42.26±2.67)g/L]and serum ionized calcium[male(1.27±0.04)mmol/L vs female(1.26±0.04)mmol/L]were observed between different genders of Chinese volunteers.However,citrate intervention led to more pronounced decrease of ionized calcium level in Chinese females compared to Chinese males[-28.68%(-20.00%--35.2%)vs-23.84%(-16.53%--29.32%),t=3.19,P ＜ 0.01].There was no differences of the levels of serum inorganic phosphate[-18.81%(-3.16%--25.09%)vs-19.23%(-1.22%--32.16%),t=0.36,P＞0.05]and albumin[-0.32%(3.27%--7.60%)vs 1.88%(6.03%--9.31%),t=0.47,P＞0.05].Independent of gender,citrate intervention resulted in an increased excretion of urine calcium in Chinese volunteers[before 0.34(0.09-0.87)vs after 0.96(0.18-1.47),t=6.66,P ＜0.01].Compared to Caucasian males,Chinese males has a higher basal level of serum ionized calcium [(1.27±0.04)mmol/L vs(1.22±0.02)mmol/L,t=3.7,P ＜0.01]and larger amplitude basal rhythm in serum albumin level[-11.72%(-5.70%--14.21%)vs-1.74%(2.43%--7.68%),t=7.43,P ＜ 0.01].Application of citrate resulted in comparable changes of serum ionized calcium [-23.84%(-16.53%--29.32%)vs-21.95%(-18.31%--30.92%)],phosphate[-19.23%(4.65%--32.16%)vs-12.68%(0.68%--42.19%)],albumin[-0.32%(1.05%--7.60%)vs-1.39%(1.87%--7.26%)]and urine calcium excretion[237.70%(11.8%-935%)vs 234.37%(5.45%-504.00%)]between Chinese and Caucasian males(t=0.32,0.03,0.25 and 0.04 respectively,P＞0.05).Serum levels of magnesium were not influenced in all volunteers during two interventions.Conclusions Independent of race and gender,the invention of citrate results in comparable changes of serum magnesium,inorganic phosphate and albumin.The effect of citrate on ionized calcium levels between genders implicates a higher risk for hypocalcemic reactions in females compared to males undergoing automatic apheresis procedures.

In order to evaluate the efficacy and side effects of the non-insulin antidiabetes medications as an adjunct treatment in type 1 diabetes mellitus (T1DM), we conducted systematic searches in MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials for randomized controlled trials published between the date of inception and March 2020 to produce a systematic review and meta-analysis. Overall, 57 studies were included. Compared with placebo, antidiabetes agents in adjunct to insulin treatment resulted in significant reduction in glycosylated hemoglobin (weighted mean difference [WMD], –0.30%; 95% confidence interval [CI], –0.34 to –0.25%; P<0.01) and body weight (WMD, –2.15 kg; 95% CI, –2.77 to –1.53 kg; P<0.01), and required a significantly lower dosage of insulin (WMD, –5.17 unit/day; 95% CI, –6.77 to –3.57 unit/day; P<0.01). Compared with placebo, antidiabetes agents in adjunct to insulin treatment increased the risk of hypoglycemia (relative risk [RR], 1.04; 95% CI, 1.01 to 1.08; P=0.02) and gastrointestinal side effects (RR, 1.99; 95% CI, 1.61 to 2.46; P<0.01) in patients with T1DM. Compared with placebo, the use of non-insulin antidiabetes agents in addition to insulin could lead to glycemic improvement, weight control and lower insulin dosage, while they might be associated with increased risks of hypoglycemia and gastrointestinal side effects in patients with T1DM.

In order to evaluate the efficacy and side effects of the non-insulin antidiabetes medications as an adjunct treatment in type 1 diabetes mellitus (T1DM), we conducted systematic searches in MEDLINE, Embase, and the Cochrane Central Register of Controlled Trials for randomized controlled trials published between the date of inception and March 2020 to produce a systematic review and meta-analysis. Overall, 57 studies were included. Compared with placebo, antidiabetes agents in adjunct to insulin treatment resulted in significant reduction in glycosylated hemoglobin (weighted mean difference [WMD], –0.30%; 95% confidence interval [CI], –0.34 to –0.25%; P<0.01) and body weight (WMD, –2.15 kg; 95% CI, –2.77 to –1.53 kg; P<0.01), and required a significantly lower dosage of insulin (WMD, –5.17 unit/day; 95% CI, –6.77 to –3.57 unit/day; P<0.01). Compared with placebo, antidiabetes agents in adjunct to insulin treatment increased the risk of hypoglycemia (relative risk [RR], 1.04; 95% CI, 1.01 to 1.08; P=0.02) and gastrointestinal side effects (RR, 1.99; 95% CI, 1.61 to 2.46; P<0.01) in patients with T1DM. Compared with placebo, the use of non-insulin antidiabetes agents in addition to insulin could lead to glycemic improvement, weight control and lower insulin dosage, while they might be associated with increased risks of hypoglycemia and gastrointestinal side effects in patients with T1DM.

Objective:To investigate the gene mutation and expression profiles in patients with acute lymphoblastic leukemia (ALL) and the effect of gene mutations on the prognosis of patients.Methods:DNA samples from 128 newly diagnosed ALL patients in the First Affiliated Hospital of Soochow University from June 2016 to June 2017 were collected. The targeted specific next-generation sequencing technology was used to analyze 51 gene mutations related to hematological malignancies, and the occurrence spectrum was described. Because the gene mutation spectrum varied with the subtype of disease, the gene mutations involved 8 types of pathways including DNA methylation, chromosomal modification, transcriptional regulation, tumor suppression, signal transduction, RNA splicing, adhesive complexes and other pathways. The effects of clinical factors and gene mutations on overall survival (OS) and relapse-free survival (RFS) were analyzed by Kaplan-Meier method and Cox regression model.Results:Of the 128 patients, the results of next-generation sequencing showed that 86 patients (67.2%) harbored at least one mutation, and 27 patients (21.1%) harbored ≥3 mutations based on the next-generation sequencing. In all ALL patients, the genes with high mutation rates were JAK (10.9%, 14/128), NOTCH1 (10.1%, 13/128), KRAS (8.6%, 11/128), SETD2 (7.0%, 9/128), CSMD1 (7.0%, 9/128), ETV6 (7.0%, 9/128), and RUNX1 (7.0%, 9/128). In B-cell acute lymphoblastic leukemia (B-ALL) patients, the genes with high mutation rates were KRAS (9.4%, 10/106), CSMD1 (7.5%, 8/106), JAK (7.5%, 8/106), PTPN11 (6.6%, 7/106), SETD2 (5.7%, 6/106), TET2 (5.7%, 6/106), TP53 (5.7%, 6/106), and PAX5 (5.7%, 6/106). While in T-cell acute lymphoblastic leukemia (T-ALL) patients, the genes with high mutation rates were NOTCH1 (54.5%, 12/22), PHF6 (27.3%, 6/22), JAK (27.3%, 6/22), RUNX1 (22.7%, 5/22), and ETV6 (18.2%, 4/22). In 128 ALL patients, the total frequency of gene mutations was 181 times. Among them, signal transduction, transcriptional regulation, tumor suppression and chromosomal modification-related gene mutations occurred more frequently, and similar phenomena were found in T-ALL and B-ALL. In terms of clinical features, male patients were more likely to present gene co-mutations( P=0.002), and mutations involved in tumor suppressor pathways were also more common in male patients ( P=0.054). The older the patient, the greater the possibility of mutations involved in transcriptional regulation and DNA methylation regulatory pathway-related genes ( P=0.067, P=0.009). T-ALL was more susceptible to have gene mutations than B-ALL ( P=0.002), and easily had gene co-mutations ( P < 0.01), and mutations mainly involved in signal transduction, transcriptional regulation, tumor suppression and chromosome modification were dominant (all P < 0.05). Cox regression univariate analysis showed that younger age of onset and allogeneic hematopoietic stem cell transplantation could significantly prolong the OS time of ALL patients ( P=0.005, P=0.003), but the difference was not statistically significant on RFS (both P > 0.05). However, 8 types of regulatory pathways were irrelevant to OS and RFS in ALL patients (all P > 0.05). The ALL patients with JAK gene mutation had short OS time ( P=0.024). Conclusions:Gene mutations are prevalent in ALL patients, the frequency spectrum varies with the subtype of disease and involves a variety of signaling pathways. Among them, signal transduction, transcriptional regulation, tumor suppression and chromosomal modification pathway-related genes have high mutation rates. The co-occurrence of gene mutations is a frequently phenomenon in ALL patients and it indicates genetic complexity and instability of ALL patients. JAK family gene mutations usually indicate poor prognosis, but the effects of other gene mutations on the prognosis of ALL need to be further explored.

Objective:To explore the correlation of serum neurogenic exosome MicroRNA-211-5p(miR-211-5p)levels and brain derived neurotrophic factor(BDNF)levels with cognitive impairment in patients with Parkinson's disease and their diagnostic value.Methods:A total of 80 patients with Parkinson's disease(PD)admitted to the Second Hospital of Jiaxing City from January 2017 to April 2018 were enrolled.According to the Montreal cognitive assessment scale, patients were divided into the cognitive impairment group(n=36)and the non-cognitive impairment group(n=44). Meanwhile, 30 healthy people who took health check-ups during the same period were selected as the control group.Exosomes were extracted from peripheral blood of subjects by using the ExoQuick kit, and the neurogenic exosomes were separated by an L1 cell adhesion molecule(L1CAM)biotinylated antibody.BDNF levels in the exosomes were measured with an enzyme-linked immunosorbent assay(ELISA), and the expression level of miR-211-5p in the exosome was determined by real-time fluorescence quantitative PCR(RT-QPCR).Results:There was a correlation between BDNF and miR-211-5p( r=-0.805, P<0.001)in serum neurogenic exosomes( r=-0.805, P<0.001). BDNF was correlated with miR-211-5p in both the PD and control groups( r=-0.785 and-0.867, P=0.002 and 0.001). The miR-211-5p level was higher and the BDNF level was lower in the PD group than in the control group(0.30±0.08 vs. 0.17±0.04, 0.55±0.06 mg/L vs. 0.75±0.06 mg/L, t=7.125 and 6.368, P=0.000 and 0.000). The BDNF level was lower(0.45±0.07 mg/L vs.0.63±0.07 6.368 and 0.75±0.08 mg/L, t=8.999 and 7.608, P=0.000 and 0.000)and the MiR-211-5p level was higher(0.36±0.07 vs. 0.24±0.05 and 0.17±0.04, t=10.923 and 7.520, P=0.000 and 0.000)in the cognitive impairment group than in the non-cognitive impairment and control groups.The receiver-operating characteristics(ROC)curve showed that the area under the curve of miR-211-5p as a measure for cognitive impairment in Parkinson's disease was 0.860(95% CI: 0.770-0.950)with a threshold of 0.32.The area under the curve of BDNF as a measure for cognitive impairment in Parkinson's disease was 0.891(95% CI: 0.822-0.961)with a threshold of 0.67.BDNF seemed to be the target gene of miR-211-5p, since the latter could inhibit BDNF expression by reducing BDNF mRNA levels. Conclusions:Human serum neurogenic exosome miR-211-5p is highly expressed in PD patients with cognitive impairment and has the potential to be used as one of diagnostic parameters for cognitive impairment in PD patients.The high expression of serum neurogenic exosome miR-211-5p may be related to the inhibition of BDNF by reducing its mRNA levels.