Objective:To explore the significance of next-generation sequencing to detect gene mutations for diagnosis of patients with unexplained pancytopenia.Methods:The next-generation sequencing was used to detect 32 common hematological tumor gene mutations in 113 patients with unexplained pancytopenia in Huazhong University of Science and Technology Union Shenzhen Hospital from February 2016 to February 2019.Results:The next-generation sequencing results showed that of the 32 genes tested in 113 patients, 69 mutations occurred in 25 genes. Among them, 105 patients (92.9%) carried at least 1 gene mutation, and the median number of mutations was 2 (1-5). Fifty-one patients were diagnosed with myelodysplastic syndrome (MDS), and the related mutant genes were detected; 62 patients were diagnosed with atypical aplastic anemia or cytopenia of unknown significance. The total effective rate of treatment for 113 patients was 65.5% (74/113).Conclusion:The next-generation sequencing helps to diagnose patients with unexplained pancytopenia, and provides targeted strategies for finding new treatments and prolonging the overall survival of patients.

Objective:To explore the correlation between the level of serum free light chain and renal function indexes in patients with diabetic kidney disease (DKD).Methods:The clinical data of 120 patients with DKD (study group) and 80 patients with simple diabetes (diabetes group) in the Beijing Sixth Hospital from October 2017 to October 2020 were retrospectively analyzed. Five ml of fasting peripheral venous blood were collected, the levels of serum uric acid, creatinine, homocysteine (Hcy) and free light chain (including free light chain κ, free light chain λ, ratio of free light chain κ and free light chain λ and total free light chain) were detected, and the estimated glomerular filtration rate (eGFR) was calculated. The correlation between total free light chain and renal function indexes in patients with DKD was analysis by Pearson method.Results:The free light chain κ, free light chain λ, ratio of free light chain κ and free light chain λ, total free light chain, uric acid, creatinine and Hcy in DKD group were significantly higher than those in diabetes group: (33.92 ± 9.06) mg/L vs. (17.65 ± 4.72) mg/L, (17.52 ± 2.83) mg/L vs. (9.81 ± 3.20) mg/L, 1.93 ± 0.23 vs. 1.80 ± 0.25, (51.44 ± 12.31) mg/L vs. (27.46 ± 7.92) mg/L, (383.69 ± 96.11) μmol/L vs. (345.93 ± 93.94) μmol/L, (117.57 ± 22.39) μmol/L vs. (75.06 ± 14.73) μmol/L and (17.64 ± 5.76) μmol/L vs. (11.66 ± 5.46) μmol/L, the eGFR was significantly lower than that in diabetes group: (103.95 ± 22.58) ml/(min·1.73 m 2) vs. (142.65 ± 26.50) ml/(min·1.73 m 2), and there were statistical differences ( P<0.01). Pearson correlation analysis results showed that serum total free light chain in patients with DKD was positively correlated with uric acid, creatinine and Hcy ( r = 0.707, 0.709 and 0.820; P<0.01), and negatively correlated with eGFR ( r = -0.730, P<0.01). Conclusions:The expression level of serum free light chain is obviously increased in patients with DKD, it has a certain correlation with renal function indexes. It can be used as one of the evaluation indicators for disease monitoring.

Objective:To observe the therapeutic effect of microwave ablation combined with anhydrous ethanol for cystic thyroid nodules.Methods:From Jan. 2019 to Dec. 2019, 56 patients with thyroid cystic nodules (≥2cm) underwent ultrasound guided thyroid cystic nodule ablation in Department of Thyroid and Breast Surgery, Nanjing Hospital of Traditional Chinese Medicine. According to different ablation methods, the patients were divided into microwave ablation combined with anhydrous ethanol group and microwave ablation group. There were 36 cases in microwave ablation combined with anhydrous ethanol group and 20 cases in microwave ablation group. The volume reduction rate of thyroid nodules, the incidence of postoperative complications and the changes of thyroid function were compared between the two groups after treatment. Statistical analysis were performed using SPSS, version 21.0, the mean±SD deviation ( ± s) was used to describe the statistics, t-test was performed, and the adoption rate of counting data (%) was expressed by χ 2 test. The difference was statistically significant with P<0.05. Results:The nodule volume reduction rates of the microwave ablation combined with anhydrous ethanol group and microwave ablation group were (49.86±6.78) % vs (22.84±1.88) %, (67.57±5.84) % vs (47.25±7.09) % and (75.70±4.51) % vs (71.14±4.65) % at 3 months, 6 months and 12 months after operation, respectively. There was significant difference between the two groups ( P<0.001) . The incidence of postoperative complications in the two groups was 38.89% and 45.00% respectively, and there was no significant difference between the two groups ( P>0.05) , and all complications were cured within 2 months. There was no significant difference in thyroid function (T3, T4, FT3, FT4, TSH) between the two groups before and 12 months after operation ( P>0.05) . Conclusions:Microwave ablation combined with anhydrous ethanol is more effective in treatment of cystic thyroid nodules (≥2cm) than microwave ablation alone. It can significantly improve patients’symptoms and nodule volume reduction, and does not affect thyroid function. It can be used as a recommended option for treatment of cysticthyroid nodules.

Objective To evaluate the value and short-term efficacy of Miccoli for cervical lymph node dissection in patients with thyroid papillary carcinoma.Methods From Mar.2017 to Jan.2018,15 patients with thyroid papillary carcinoma received cervical lymph node dissection in Department of Thyroid and Breast Surgery of Nanjing Chinese Traditional Medicine Hospital,and they were divided into open surgery group (8 cases) and Miccoli surgery group (7 cases).The operation time,incision length,intraoperative blood loss,total number of dissection lymph nodes,incidence of postoperative complications and length of hospital stay between the two groups were analyzed.These groups were followed up for 6 months after surgery,in terms of neck comfort,incidence of enlarged lymph nodes in the cervical region and blood thyroglobulin level.Results There was no obvious difference between the two groups in intraoperative blood loss(P=0.651),total number of dissection lymph nodes(P=0.887),the incidence of postoperative complications(P=0.52),incidence of intumescent lymph node(P=1.000) and blood thyroglobulin level(P=0.826) after 6 months.The operation time of Miccoli group was longer than that of the open group(P=0.001),but the incision length(P=0.001),length of hospital stay(P=0.001)and postoperative neck comfort(P=0.001)were superior to those of the open surgery group.Conclusion Miccoli operation can be used to perform cervical lymph node dissection for PTC patients,which can achieve the curative effect of open surgery,without increasing the risk of postoperative complications,and the postoperative neck comfort was better.

Objective:To investigate the clinical effect of modified arrow flap for nipple reconstruction.Methods:From Jan. 2018 to Oct. 2019, 10 patients in Nanjing Hospital of Traditional Chinese Medicine who received the modified arrow flap for nipple reconstruction were collected. The rate of nipple retraction, patient satisfaction and the incidence of local flap complications were evaluated.Results:The operation time of 10 patients was 13-18 minutes, and the average operation time was (15.10±1.52) minutes. All patients were followed up for 3-24 months, with an average follow-up time of (14.60±1.07) months. The retraction rate of nipple height was 23.5%-33.2%, the average retraction rate of nipple height was (28.53±3.02) %, the diameter retraction rate of nipple was 7.1%-10.5%, and the average diameter retraction rate of nipple was (8.92±1.05) %. The patients’ satisfaction was 96%-100%, with an average of (97.9±1.60) %. None of the 10 patients had complications such as bad wound healing, incision dehiscence or flap necrosis.Conclusion:The design of the improved arrow flap is simple, the operation is simple, the curative effect is satisfactory, and the incidence of postoperative complications is low.

Objective:To study the role of miR-155 in the differentiation of papillary thyroid carcinoma (PTC) cells, and to explore the possible mechanism.Methods:Human miR-155 analogues were constructed and transfected into PTC BCPAP cells in vitro. CCK8 test and Transwell test were used to observe the changes of cell proliferation and invasiveness. The miR-155 was transfected into BCPAP cells in vitro and the protein background and phosphorylation expression of MAPK pathway were detected by Western blot. ERK pathway inhibitor U0126 was given to observe whether it could reverse the abnormal proliferation and invasion of thyroid cancer cells caused by over-expression of miR-155.Results:The proliferation of BCPAP cells was detected by CCK8 test 48 hours after overexpression of miR-155, and the invasiveness of thyroid cancer cells was significantly enhanced by Transwell test 48 hours after overexpression of miR-155 ( P<0.05) ; Western blot method found that the expression of JNK, ERK and p38 in MAPK signal pathway was significantly up-regulated ( P<0.05) . At the same time, the expression of p-ERK protein in the cells was increased significantly ( P<0.05) . The expression of p-ERK in the cells treated with ERK pathway inhibitor U0126 and miR-155 was significantly lower than that in the miR-155 group ( P<0.05) . By detecting the proliferation and invasion of cells in each group, we found that the U0126 could reverse the proliferation and invasion promoting effect caused by miR-155. Conclusion:miR-155 can promote the proliferation and invasion of PTC BCPAP cells by activating the ERK pathway of MAPK pathway, which provides a potential target for the treatment of thyroid cancer.

ObjectiveTo investigate the promotional effect of astragaloside on the repair and healing of chronic non-healing wounds and its mechanism. MethodA total of 60 male SD rats were constructed with full-layer skin defect wounds on the back， and except for the control （Con） group， the rest were constructed with non-healing wounds， which were then randomly divided into the sham-operation （sham） group， the low-dose astragaloside group， the high-dose astragaloside group， the astragaloside + LY294002 ［phosphatidylinositol 3-kinase （PI3K） inhibitor］ group， and the astragaloside + EX527 ［silencing regulatory protein 1 （SIRT1） inhibitor］ group. The percentage of wound area in each group was observed on the 2nd， 4th， 6th， and 8th days after wound molding. Collagen type Ⅰ alpha 1 （COL1A1） and alpha smooth muscle actin （α-SMA） expressions in the wound tissue were detected by immunofluorescence. Hematoxylin and eosin （HE） staining was performed to determine the pathological structure of the wound. The mRNA expression of inflammatory factors in the wound was measured by real-time polymerase chain reaction （Real-time PCR）， and the expression of proteins related to the SIRT1/ nuclear factor （NF）-κB and PI3K/protein kinase B （Akt） signaling pathways in the wound was tested by Western blot. ResultCompared with the sham group， the percentage of postoperative wound area of rats in both low-dose and high-dose astragaloside groups gradually decreased with time， and the efficacy of the high-dose astragaloside group was better. Compared with the Con group， the fluorescence intensity of COL1A1 in wound tissue of the sham group decreased， while the expression of α-SMA increased. The epithelial tissue was severely damaged， with an increase in the thickness， and a large number of inflammatory cells were seen in the infiltration. The mRNA expression of tumor necrosis factor （TNF）-α， interleukin （IL）-1β， IL-6， and inducible nitric oxide synthase （iNOS） was elevated. The protein expression of NF-κB p65， p-PI3K/PI3K， and p-Akt/Akt was elevated， while SIRT1 expression was decreased （P<0.05）. Compared with the sham group， the fluorescence intensity of COL1A1 and α-SMA increased after astragaloside treatment. The number of epithelial cells increased， and the thickness decreased. The inflammatory cells decreased， and the amount of collagen increased. The mRNA expression of TNF-α， IL-1β， IL-6， and iNOS was decreased， and the protein expression of NF-κB p65， p-PI3K/PI3K， and p-Akt/Akt was decreased. SIRT1 was elevated， and the effect was better in the high-dose astragaloside group （P<0.05）. Compared with the high-dose astragaloside group， inhibition of the PI3K/Akt and SIRT1 pathways by LY294002 and EX527 prevented the therapeutic efficacy of astragaloside on chronic non-healing wounds. ConclusionThe topical application of astragaloside significantly promotes the healing of chronic non-healing wounds in rats， and the mechanism may be related to the activation of the PI3K/Akt pathway and the SIRT1/NF-κB pathway.

An increased level of reactive oxygen species is a key factor in neuronal apoptosis and epileptic seizures. Irisin reportedly attenuates the apoptosis and injury induced by oxidative stress. Therefore, we evaluated the effects of exogenous irisin in a kainic acid (KA)-induced chronic spontaneous epilepsy rat model. The results indicated that exogenous irisin significantly attenuated the KA-induced neuronal injury, learning and memory defects, and seizures. Irisin treatment also increased the levels of brain-derived neurotrophic factor (BDNF) and uncoupling protein 2 (UCP2), which were initially reduced following KA administration. Furthermore, the specific inhibitor of UCP2 (genipin) was administered to evaluate the possible protective mechanism of irisin. The reduced apoptosis, neurodegeneration, and spontaneous seizures in rats treated with irisin were significantly reversed by genipin administration. Our findings indicated that neuronal injury in KA-induced chronic epilepsy might be related to reduced levels of BDNF and UCP2. Moreover, our results confirmed the inhibition of neuronal injury and epileptic seizures by exogenous irisin. The protective effects of irisin may be mediated through the BDNF-mediated UCP2 level. Our results thus highlight irisin as a valuable therapeutic strategy against neuronal injury and epileptic seizures.
Rats ; Animals ; Kainic Acid/toxicity* ; Brain-Derived Neurotrophic Factor/metabolism* ; Fibronectins/metabolism* ; Hippocampus/metabolism* ; Rats, Sprague-Dawley ; Epilepsy/metabolism* ; Seizures/prevention & control*

Fibroblast growth factor receptors (FGFRs) have emerged as promising targets for anticancer therapy. In this study, we synthesized and evaluated the biological activity of 66 pyrazolo[3,4-