Objective To investigate the early diagnostic value of cornel confocal microscopy for the screening of small neuropathy in elderly patients with type 2 diabetic mellitus.Methods In the prospective study,96 elderly patients with diabetes as study group and 46 patients with non-diabetes as the control group were continuously collected from our hospital endocrinology and ophthalmology out patients during May 2014 to February 2016.The 96 cases of type 2 diabetes were subdivided into 47 patients with diabetic peripheral neuropathy (DPN)and 47 patients with nowdiabetic peripheral neuropathy(non DPN).Results The diabetes course was shorter in non-DPN group than in DPN group(P=0.000).The levels of glycosylated hemoglobin and urine albumin were lower in the nonDPN than in the DPN(P =0.072,0.007,respectively).The corneal nerve fiber density was lower in the DPN group than in NDPN group (P =0.000).Corneal nerve fiber density was higher in control group than in DPN and NDPN group.The differences in number of corneal nerve fibers showed no statistical significance between DPN and NDPN group (x2 =2.391,P =0.314).But the number of corneal nerve fibers was significant less in DPN and NDPN group than in control group(x2 =16.014,P =0.000).The negative correlation was found between the course of disease and corneal fibrous density by using single factor linear regression analysis.The number of corneal nerve fibers was lower in smoking group than in non-smoking group(P=0.003).The multiple linear regression analysis showed that duration of diabetes was a risk factor for diabetic neuropathy.Conclusions In some elderly diabetic patients with non-neuropathy,corneal nerve fiber density and number have been significantly decreased before nerve conductive velocity is reduced.Therefore,corneal confocal microscopy can be used to detect and diagnose small diabetic neuropathy in elderly patients with diabetes mellitus.

Objective:To study the effects of Docetaxel o n the proliferation of adenoid cystic carcinoma SACC-83 cells of salivary gland. Methods:The inhibitory effects of Docetaxel on the proliferatio n of SACC-83 cells were investigated with cell counting, soft agar clonogenic a ssay, and flow cytometry. Results:With the exposure time of 24, 48 or 72 h the IC 30(nmol/L) of Docetaxel was 1.39,1.26 and 0.47, the I C 50(nmol/L) 13.02, 3.34 and 1.26 respectively; the relative antitumor acti vity (RAA) of the drug against SACC-83 cells was 330, 1 289 and 3 426 respectiv ely. After the cells had been treated for 72 h, the percentages of G 1, S and G 2 phase cells in the cell cycle in the control group were 73.8,19.8 and 6.4, in IC 30 group 65.0, 29.5 and 5.5, in IC 50 group 57.6,42.4 and 0, res pectively. The clonogenesity (%) in control, IC 30 and IC 50 groups we re 36.0?0.5,8.3?2.5 and 0.5?0.3 respectively. Conclusion:Doc etaxel may inhibit the proliferation of SACC-83 cells in a dose and time depend ent way.

OBJECTIVE: To explore application value and efficacy of personalized osteotomy in primary total knee arthroplasty (TKA) for severe varus knee osteoarthritis. METHODS: From June 2018 to January 2020, 36 patients (49 knees) with severe varus knee osteoarthritis were treated, including 15 males (21 knees) and 21 females (28 knees), aged from 59 to 82 years old with an average of (67.6 ± 6.5) years old;the course of disease ranged from 9.5 to 20.5 years with an average of (15.0 ± 5.0) years;11 patients (15 knees) with Kellgren-Lawrence grade Ⅲ and 25 patients (34 knees) with grade Ⅳ according to Kellgren-Lawrence grading. According to AORI clsssification of tibial bone defects, 8 patients (15 knees) were typeⅠTa and 16 patients (18 knees) were typeⅡ T2a. All patients' femor-tibial angle (FTA) was above 15°, and received primary TKA with personalized osteotomy. Thirty-three patients (45 knees) were treated with posterior-stabilized (PS) prostheses, 13 patients (15 knees) with PS prostheses combined with a metal pad and extension rod on the tibial side, and 3 patients (4 knees) with legacy constrained condylar knee (LCCK) prostheses. FTA, posterior condylar angle (PCA), medial proximal tibial angle (MPTA) and lateral distal femoral angle (LDFA) before and after operation at 1 month were measured and compared by using picture archiving and communication systems (PACS). Recovery of lower limbs before and after operation at 12 months was evaluated by American Knee Society Score(KSS), and complications were observed and recorded. RESULTS: All 36 patients (49 knees) were followed up from 15 to 40 months with an average of (23.46±7.65) months. FTA, MPTA were corrected from preoperative (18.65±4.28)° and (83.75±3.65)° to postoperative (2.35±1.46)° and (88.85±2.25)° at 1 month, respectively (P<0.001). PCA was decreased from (2.42±2.16)° before operation to (1.65±1.35)° at 1 month after operation, LDFA improved from (89.56 ± 3.55)° before operation to (91.63±3.38)° at 1 month after operation (P<0.05). KSS increased from (67.58±24.16) before opertion to(171.31±15.24) at 12 months after operation, 14 patients (19 knees) got excellent result, 19 (26 knees) good, and 3 (4 knees) fair. CONCLUSION Personalized osteotomy is helpful for recovery of axial alignment of lower limbs and correct placement of prosthesis, could effectively relieve pain of knee joint, recover knee joint function.
Male ; Female ; Humans ; Child ; Adolescent ; Young Adult ; Adult ; Arthroplasty, Replacement, Knee ; Osteoarthritis, Knee/surgery* ; Retrospective Studies ; Knee Joint/surgery* ; Osteotomy ; Tibia/surgery*

Objective To study the absorption features of isoimperatorin in intestine of rat.Methods Establish a single-pass intestinal perfusion model of rat,take phenolsulfonphthalein as a marker for the detection of isoimperatorin concentration from crude extracts of notopterygium,and observe the absorption features of isoimperatorin and its reference substance in intestine of rat.Results The content of isoimperatorin in crude extracts was (0.43±0.02)% (n=5).The absorption parameters of duodenum,jejunum,ileum,colon were (5.28±0.82),(4.47±0.56),(4.17±0.94),(4.32±0.68)×10-5 cm·s-1,respectively.There were no significant differences among them (P>0.05).Compared with the reference substance of isoimperatorin,crude extracts showed better absorption features.Conclusion Isoimperatorin from crude extracts have better characteristics of absorption.This study can provide theoretical basis for design of notopterygium oral formulation.

Objective:To explore whether antioxidant coenzyme Q10 (CoQ10) is involved in the regulation of renal injury induced by diquat poisoning (DQ) in rats through anti-oxidative stress and inhibition of interleukin (IL)-17 and nuclear factor kappa-B (NF-κB) signaling pathway, and whether this mechanism is related to alleviating mitochondrial dysfunction.Methods:The expressions of NF-κB inhibitory protein α (IKB-α), phosphorylated nuclear factor κB (P-NF-κB), JNK-related leucine zipper protein (JLP) and neuroprotective protein PTEN-induced putative kinase 1(PINK1) pathway proteins were detected in vivo and in vitro. Biochemical detection of renal injury markers and inflammatory cytokines: serum urea nitrogen (BUN), serum creatinine (Cr), Cystatin C (CysC), renal injury molecule 1, Malondialdehyde, Supemxidedismutase (SOD), neutrophil gelatinase-associated lipocalin (NGAL), etc. Renal pathology HE staining was used to observe the degree of renal injury and pathological score under light microscope. The expression of reactive oxygen species (ROS) was detected by immunofluorescence. CCK-8 experiment was used to observe the level of cell proliferation after administration.Results:In vivo experiment, the indexes of renal function injury (Cr, BUN, CysC, NAGL, KIM-1) in plasma and kidney samples were significantly increased after 72 h of exposure in DQ group, and there were significant histopathological changes and pathological scores increased. In vitro experiment HK-2 cells were exposed to DQ for 48 h, and the cell viability decreased by half. After exposure to DQ, serum SOD decreased, MDA increased, and the immunofluorescence value of ROS in renal tissue increased. Intervention with CoQ10 can alleviate the pathological damage induced by DQ in rats, enhance the vitality of HK-2 cells, alleviate renal injury and reduce the level of oxidative stress. In addition, the expression of pro-inflammatory cytokines (IL-6, TNF-α and IL-17) increased in DQ group in vivo, the expression of P-NF-κBp65 protein in DQ group in vivo and HK-2 cell DQ group in vitro increased significantly, the expression of mitochondrial dysfunction index PINK1 protein increased significantly, and the expression of JLP protein and IκB-α protein decreased significantly. After intervention with CoQ10, the expression of P-NF-κBp65 protein and PINK1 can be decreased, while the expression of IκB-α protein can be increased and the degradation of JLP could be alleviated, and CoQ10 could improve the mitochondrial dysfunction after DQ poisoning.Conclusions:CoQ10 can alleviate the kidney injury induced by DQ poisoning in rats, and its mechanism may be related to the fact that CoQ10 regulates the expression of IL-17 and NF-κB signaling pathway through anti-oxidative stress, and further improves mitochondrial dysfunction.

Aspergillus niger is an important industrial strain which has been widely used for production of enzymes and organic acids. Genome modification of A. niger is required to further improve its potential for industrial production. CRISPR/Cas9 is a widely used genome editing technique for A. niger, but its application in industrial strains modification is hampered by the need for integration of a selection marker into the genome or low gene editing efficiency. Here we report a highly efficient marker-free genome editing method for A. niger based on CRISPR/Cas9 technique. Firstly, we constructed a co-expression plasmid of sgRNA and Cas9 with a replication initiation region fragment AMA1 (autonomously maintained in Aspergillus) by using 5S rRNA promoter which improved sgRNA expression. Meanwhile, a strain deficient in non-homologous end-joining (NHEJ) was developed by knocking out the kusA gene. Finally, we took advantage of the instability of plasmid containing AMA1 fragment to cure the co-expression plasmid containing sgRNA and Cas9 through passaging on non-selective plate. With this method, the efficiency of gene editing reached 100% when using maker-free donor DNA with a short homologous arm of 20 bp. This method may facilitate investigation of gene functions and construction of cell factories for A. niger.
Gene Editing ; Aspergillus niger/genetics* ; CRISPR-Cas Systems/genetics* ; Plasmids/genetics*