Objective:To establish a rat model of diabetic microangiopathopathy and simulate the biochemical and pathological changes of diabetic retinal and renal microangiopathopathy.Methods:Forty healthy male Sprague-Dawley rats were randomly divided into blank group and model group (10 and 30 rats, respectively). After the rats in blank group and model group were fed ordinary diet and high-fat and high-sugar diet for 5 weeks, respectively, the rats in model group were injected with 1% streptozotocin (STZ) through the abdominal cavity at the dose of 35 mg/kg to establish a type 2 diabetes model. After modeling, the rats were continuously fed until the 10th week (4 weeks after modeling), the general conditions of the rats were observed, and samples were collected for follow-up experiments. Serum creatinine (CREA), triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL-C), low density lipoprotein (LDL-C), microalbuminuria, urinary creatinine (UCr) and urine sugar were detected. Calculate the kidney index and microalbumin/urinary creatinine ratio (UACR). Optical coherence tomography angiography (OCTA) was used to observe the vascular changes and non-perfusion area of retinal superficial capillary plexus. The morphological and structural changes of kidney and retina were observed by hematoxylin-eosin and periodate Scheff staining. The expression of nerve fibers and nucleus of Müller cells in rat retina was observed by immunofluorescence staining. Ultrastructural results of retina were observed by transmission electron microscope. Independent sample t test was used for comparison between groups. Results:Four weeks after modeling, compared with blank group, the body weight of rats in model group was significantly decreased, and random glucose was significantly increased, with statistical significance ( t=5.755, -51.291; P<0.05). Renal index, urinary glucose and UACR were significantly increased, while UCr was significantly decreased, with statistical significance ( t=10.878, 137.273, 3.482,-6.110; P<0.05). CREA decreased, TG, TC, HDL-C, LDL-C increased, and the differences were statistically significant ( t=-28.012, 33.018, 118.018, 13.585, 16.480; P<0.05). OCTA examination showed that there was no perfusion area of shallow retinal capillaries. The optical microscope showed that the inner boundary membrane of retina in model group was swollen and thickened, the surface was uneven, the inner and outer nuclear layer cells were disordered and the density decreased. Glomerular congestion was accompanied by cortical tubular epithelial swelling, widening of the mesangial area, and thickening of the basement membrane. The results of immunostaining showed that the inner and outer plexiform layers of the retina showed lamellar strong green fluorescence expression, and the inner and outer nuclear layers showed scattered dot green fluorescence expression. Transmission electron microscopy showed that the basal membrane of retinal microvessels in model group was slightly thickened, vascular endothelial cells edema, endothelial nucleus and perinucleus contraction, nuclear membrane contraction, mild mitochondrial swelling, vacuolation. Conclusion:High-glucose and high-fat feeding plus a single intraperitoneal injection of STZ 35 mg/kg can successfully establish a microangiopathic model of type 2 diabetes.

Objective:To investigate the feasibility, effectiveness and safety of indocyanine green (ICG) navigation in the surgical resection of abdominal wall endometriosis (AWE).Methods:Seven women undergoing surgery for AWE in First Affiliated Hospital of Sun Yat-sen University (from July 1, 2021 to October 1, 2021) were collected. After exposure of the focus, ICG were used intravenously (0.25 mg/kg) as fluorescent dye for the intraoperative evaluation of AWE vascularization. Resection of the AWE was guided by direct visualization of the focus under standard laparoscopy with a near-infrared (NIR) camera head. Surgical margin around the AWE (3, 6, 9 and 12 point) and the margin under the focus were obtained for postoperative pathological examination of endometriosis. Time from injection to fluorescence visualization, the proportion of fluorescence visualization, time of fully resection of AWE, side effects related to the use of ICG, perioperative complications as well as the pathological result of the surgical margins were recorded.Results:ICG fluorescence of the AWE were seen in 5 patients (5/7). The mean time from injection to fluorescence visualization was (46.7±9.8) s. The mean time of fully resection of AWE was (16.4±7.0) minutes. There were no side effects related to the use of ICG. The rate of class-A wound healing was 7/7. All of the surgical margins were confirmed endometriosis-negative by postoperative pathological examination.Conclusion:ICG fluorescence visualization could conduct accurate resection of AWE, which is clinically safe and effective.

Objective To investigate the clinical and biological features of patients with mixed﹣phenotype acute leukemia (MPAL). Methods The clinical data of 24 de novo adult patients with MPAL who were admitted to Fujian Medical University Union Hospital from January 2012 to October 2018 were retrospectively analyzed. These patients were diagnosed according to the World Health Organization (WHO) 2016 criteria. The clinical and biological characteristics of the patients were analyzed by morphological and cytochemical staining, immunophenotyping, cytogenetics and molecular biology. Results Of the 24 patients, 16 were male and 8 were female, and the median age of the patients at diagnosis was 27 years old (5-66 years old). The average blasts of bone marrow were (57.41 ±23.20)% . Thirteen cases (54.2% ) were diagnosed as MPAL morphologically, while 5 cases (20.8% ) were diagnosed as acute myeloid leukemia (AML), 5 cases (20.8%) were diagnosed as acute lymphoblastic leukemia (ALL) and 1 case (4.2%) was inconclusive. Eighteen patients (75.0%) co﹣expressed B﹣lymphoid and myeloid markers, while 5 patients (20.8%) with T﹣lymphoid and myeloid markers and 1 patient (4.2%) with B﹣lymphoid and T﹣lymphoid markers, respectively. The positive rate [median (range)] of CD38, HLA﹣DR and CD34 was 90.5% (0.1%-99.7%), 90.1% (1.1%-98.8% ) and 81.3% (0.1%-97.8%), respectively. Eighteen cases underwent chromosome examination, of which 5 cases carried with t(9;22)(q34;q11), 3 cases with t(v;11q23.3), 2 cases with complex karyotypes, and 2 cases with t(9;22)(q34;q11) and complex karyotypes, respectively. Twenty﹣one cases underwent genetic examination, of which 6 cases were positive for BCR﹣ABL, 3 cases were positive for MLL, 1 case was positive for MLL and BCR﹣ABL, 1 case was positive for BCR﹣ABL and TP53, and 1 case was positive for PHF6 and ASXL1 respectively. Of the 24 patients, 7 refused chemotherapy and 17 received induction chemotherapy. Of the patients receiving chemotherapy, 9 cases achieved complete remission (CR), 1 case was partial remission (PR), and 7 cases were not relieved (NR). In 11 patients treated by ALL﹣type induction regimen and 6 patients treated by ALL and AML﹣type induction regimen, 8 cases and 1 case achieved CR, the difference in CR rate was statistically significant (P<0.05). In 6 patients with Philadelphia chromosome (Ph) positive and 11 patients with Ph negative, 1 case and 8 cases achieved CR, the difference in CR rate was statistically significant (P<0.05). The median follow﹣up time was 5.5 months (0-36 months). The 3﹣year overall survival (OS) rate was 17.5% and the median OS time was 6 months. The 3﹣year OS rates in the allogeneic hematopoietic stem cell transplantation and non﹣transplanted groups were 75.2% and 0, respectively, and the median OS time was not reached and 4 months (P< 0.05). Conclusions MPAL is rare, it mostly co﹣expresses lymphoid and myeloid antigens and shows a much higher incidence of CD34, CD38 and HLA﹣DR. MPAL is often associated with Ph positive and complex karyotypes. MPAL has a low remission rate and poor prognosis, and a reasonable and effective treatment plan should be further explored.

Objective To explore the clinical features of chronic myelogenous leukemia (CML) combined with solid malignant neoplasms. Methods The clinical data of 8 CML patients with solid malignant neoplasms who were admitted to the Affiliated Tumor Hospital of Zhengzhou University, the Central Hospital of Nanyang City, the First Affiliated Hospital of Science and Technology University of Henan, and the Central Hospital of Xinxiang City from August 2006 to August 2018 were analyzed retrospectively. The clinical features, treatment and prognosis of the patients were summarized with the review of literature. Results Among the 8 patients, 3 were male and 5 female, aged 40-76 years, with a median of 50 years old. Seven cases were in CML chronic phase, and 1 was in accelerated phase. Seven patients were treated with tyrosine kinase inhibitor (TKI), and only 1 patient was treated with hydroxyurea. In 8 patients, two cases presented with synchronous multiple primary cancer (SMPC), 6 cases presented with heterochrony multiple primary cancer (HMPC). two patients received the operation, 1 patient received the operation and chemotherapy, 4 patients received chemotherapy, and 1 patient received the isotope treatment. One SMPC patient died and another one was under treatment, and 6 HMPC patients were under treatment. ConclusionsThe relationship between CML and solid malignant neoplasm is under discussion, but patients with CML and solid malignant neoplasm are not unusual. Clinicians should raise awareness to avoid misdiagnosis. The treatment should follow the two main lines that are comprehensive treatment and individualized treatment.

Objective: To investigate the differences in frequency and function of natural killer cells (NK) between chronic hepatitis B (CHB) and acute hepatitis B (AHB). Methods: Patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of NK, CD56^dimNK, CD56^brightNK and the expression of functional molecules IFNAR2 and NKp46 on the surface of NK cells were detected respectively among patients with CHB in IA phase, patients with AHB, and those recovered from AHB. At the same time, their correlations with ALT, HBV DNA and HBV markers were analyzed. Results: Between IA and AHB, the frequencies of NK cells and NKp46^dim NK cells in AHB cases were significantly lower than those in IA cases, but the frequency of NKp46^high NK cells in AHB was higher than that in IA. For patients who recovered from AHB, the frequency of NK cells and NKp46^dim NK cells increased; the varied ranges of frequencies of CD56^dimNK, IFNAR2⁺ NK and NKp46⁺ NK cells were on the rise, while the frequency of NKp46^high NK cells decreased after the recovery from AHB, and the varied ranges of CD56^brightNK and IFNAR2MFI, NKp46MFI decreased. In AHB, HBVDNA loads were positively correlated with ALT levels. Before and after the recovery of AHB: ΔHBV DNA and ΔALT, Δ NK/LY (%) were positively correlated; ΔALT and ΔNKp46^highNK/NK(%), ΔNKp46MFI, ΔIFNAR2MFI were positively correlated. Conclusions In CHB immune active phase, the activity of peripheral blood NK cells was too weak to remove the virus, but NK cells play an important role in eliminating the viruses and mediating liver tissue inflammation in AHB.

Objective: To explore the persistent viral response rate (SVR) in patients with refractory chronic hepatitis C after interferon (IFN) (peginterferon 360 μg qw) and ribavirin (PR) therapy failure. The SVR of patients with refractory chronic hepatitis C was improved by PR combined with direct antiviral agents (DAA) and proper extension of the course of therapy was applied. Methods: Seventeen cases of refractory chronic hepatitis C after IFN(peginterferon 360 μg qw) and ribavirin therapy failure were given PR combined with DAA treatment. The side effects were observed and corresponding adjustments were made on drug dosage, and SVR was recorded. Results: The 17 cases completed the whole course of treatment with PR combined with DAA for 24 weeks. All the 17 patients obtained rapid viralogical response (RVR) and SVR. After treatment, the SVR rate was 100% in patients including those with virologic relapse, retreated or previously non-responsive patients with refractory chronic hepatitis C. The adverse reaction of PR combined with DAA 24 weeks was generally mild. Conclusions The use of PR combined with DAA re-treatment in patients with refractory chronic hepatitis C can achieve SVR and shorten the treatment time. PR combined with DAA re-therapy is one of effective treatments to improve the rate of sustained viral response in patients with refractory chronic hepatitis C.

Objective: To investigate the differences in function of plasmacytoid dendritic cells (pDC) and CD4⁺ T helper cells (CD4⁺ Th cells) between acute hepatitis B (AHB) and chronic hepatitis B (CHB). Methods: In this study, patients with AHB and those with CHB in immune active (IA) phase were enrolled. The frequencies of pDC, CD86⁺ pDC, CD4⁺ T cells and their subsets, surface functional molecules were detected respectively among patients with chronic HBV infection in IA phase, patients with AHB, those recovered from AHB. Meanwhile, their correlations with ALT, HBV DNA and HBV markers were analyzed. Results: The ALT level in AHB was significantly higher than that in IA, and inflammation was more obvious in AHB. Between IA and AHB, CD86⁺ pDC frequency and the mean fluorescence intensity of functional molecule CD86 (CD86MFI) were higher in IA than those in AHB, but the frequency of CD4⁺ T cells in AHB was higher than that in IA. For patients who got over AHB, the frequency of CD86⁺ pDC increased; Th1 were on the rise, while the frequencies of CD4⁺ T and Th2 decreased after the recovery of AHB, and Th2 / Th1 ratio decreased..In AHB, HBVDNA loads were positively correlated with ALT levels and Th2 frequencies. Conclusions In CHB immune active phase, CD86⁺ pDC with stimulating function played an important role, but the cellular immune response of CD4⁺ T cells decreased. In AHB inflammatory stage, CD4⁺ T cells played a strong cellular immune response, which result ed in viral clearance. Th2 cells regulation of CD4⁺ T cells played a dominant role, which was involved in the inflammatory response, and the cytotoxic role of Th1 cells during the recovery period was dominant, playing a strong cellular immune response, then the virus were completely eliminated.

Objective To compare diagnostic values of the 2015 American Thyroid Association (ATA) Management Guidelines for Adult Patients with Thyroid Nodules and Differentiated Thyroid Cancer with the thyroid imaging reporting and data system (TI-RADS) for sonographic malignancy risk stratification of thyroid nodules.Methods From November 2011 to December 2015,485 thyroid nodules in 331 patients (mean age,42.9 years± 10.4)were included in this study.Characteristics includingsize,composition,shape(nonparallel or parallel),margin,echogenicity,calcifications and extrathyroidal extension of thyroid nodules were evaluated.Every nodule was stratificated by criteria set by TI-RADS and ATA guidelines,and malignant rate of each risk stratification were calculated and analysed.With pathology as the gold standard,different cutoff were taken to diagnose malignant nodules,and the sensitivity,specifity,positive predictive value,negativepredictive value and accuracy of the two methodologies were calculated at each cutoff.And the two methodologies were evaluated and measured by ROC curve.Finally their Kappa value were calculated at the best cutoff.Results Of the 485 thyroid nodules,96 were benign and 389 were malignant.The malignancy rates under TI-RADS category 2,3,4a,4b,4c,and 5 nodules were 0,12.0％ (3/25),22.2％ (10/45),29.8％ (14/47),99.2％ (261/363) and 100％ (101/101).Malignancy rates under ATA guidelines of benign,very low,low,intermediate,and high suspicion for malignancy were 0,12.5％ (1/8),16.1％ (10/62),27.7％ (13/47),and 99.2％ (365/368).There were significant differences inside each patterns (P ＜ 0.01) respectively and high correlation between risk stratification with TI-RADS (r=0.70) and ATA guidelines (r=0.83).Areas under the ROC curve of the TI-RADS and ATA guidelines classifications were 0.966 and 0.959.Best cut-off point for diagnosing malignant by TI-RADS and ATA guideline classifications were ≥ 4c and ≥ high suspicion,and at that point,diagnostic value of TI-RADS and ATA guidelines were nearly the same(sensitivity,93.1％vs 93.8％;specificity,97.9％ vs 96.9％;PPV,99.5％ vs 99.2％;NPV,75.7％vs 79.5％;and accuracy,94.0％vs94.4％),and there was no significant differences (P=0.50,P=0.50,P=0.50,P=0.53,P=0.55),Kappa=0.97.Conclusions Both TI-RADS and the ATA guidelinesprovide effective malignancy risk stratification for thyroid nodules.The diagnosticvalue of TI-RADS when considering ≥ 4c and ATA guidelines when considering ≥ high-suspicion nodules as malignant were nearly the same and both high.

Objective To explore the effect of inhibition of miR-214 expression on the proliferation of hepatocellular carcinoma cells via regulation of E2F3 expression.Methods The expression of miR-214 in SMMC-7721, HepG2, SK-Hep-1 and Huh 7 cells was examined by RT-PCR.Hepatocellular carcinoma cells were transfected with miR-214 NC and miR-214 mimics with liposomes.The expression of miR-214 was detected by RT-PCR.The cell viability was detected by MTT assay.Cell apoptosis was detected by Hoechst staining.Cell cycle was detected by flow cytometry.Western blot, RT-PCR and dual luciferase reporter gene assay were used to detect whether E2F3 was a downstream target gene of miR-214.Results The expression of miR-214 in SMMC-7721, HepG2, SK-Hep-1 and Huh 7 cells was 0.83±0.08, 0.32±0.03, 0.33±0.03, and 0.08±0.01, respectively.The expression of miR-214 in the HepG2 cells was the lowest, so HepG2 cells were selected as the subsequent experimental cell line.Compared with the miR-214 NC group, the expression of miR-214 (0.65±0.06 vs.0.14±0.01) was up-regulated, the cell viability (0.35±0.03 vs.0.69±0.06) was decreased, cell apoptosis rate [(36.37±3.43)% vs.(3.74±0.34)%] was increased, the G1 phase of the cell cycle (57.79±5.78 vs.45.319±4.53) was prolonged, the expression of E2F3 protein (0.23±0.02 vs.0.98±0.09) and mRNA (0.24±0.02 vs.0.99±0.10) was significantly down-regulated in the miR-214 mimics group (P<0.01).Conclusion miR-214 mimics suppress the HepG2 cell proliferation via targeted down-regulation of E2F3 expression.

Objective To compare the efficacy and safety of standard or reduced doses of bortezomib combined with adriamycin and dexamethasone in patients with multiple myeloma (MM).Methods Fifty-two newly diagnosed,refractory and relapsed patients received bortezomib [1.3 mg/m2 (standard dose group,26 patients) or 1.0-1.1 mg/m2 (reduced dose group,26 patients) on day 1st,4th,8th and 11 th],and adriamycin (10 mg/m2) plus dexamethasone (40 mg/d) on day 1 st-4th,and were treated for 1-6 courses.Adverse events were graded and compared.Results After treatment,the overall response rate of standard dose group [80.8％(21/26)] and reduced dose group [88.5％(23/26)] had no significant difference (P =0.739).The rate of neutropenia and thrombocytopenia in two groups had no significant difference[23.1％(6/26) vs.15.4％(4/26),P=0.281 ; 11.5％(3/26) vs.7.7％(2/26),P=0.620].The rate of Ⅲ-Ⅳ grade peripheral nerve disease,herpes zoster,lack of power and abdominal distension in standard dose group were significantly higher than those in reduced dose group [15.4％(4/26) vs.3.8％(1/26),P =0.038;26.9％(7/26) vs.7.7％(2/26),P =0.029;38.5％(10/26) vs.15.4％(4/26),P=0.045; 19.2％(5/26)vs.3.8％ (1/26),P =0.028].Conclusion Reduced dose of bortezomib appears to result in similar overall response rate,but better tolerance and safety compared with standard dose.