A 65-year-old male was admitted to Peking Union Medical College Hospital. The patient had intermittent fever for 2 months with a maximum body temperature of 39.3 ℃ and elevated serum creatinine levels for 1 week. He had no other suggestive symptoms or positive signs. Laboratory test results suggested acute kidney injury and a sharp elevation in serum lactic dehydrogenase levels. Abdominal enhanced computed tomography (CT) revealed multiple low-density lesions, and further biopsy pathology demonstrated chronic inflammation. Thereafter, positron emission tomography (PET)/CT showed abnormally elevated uptake value for the bones throughout the entire body, in addition to the liver and brain. Repeated bone marrow biopsy finally confirmed metastatic bone cancer, which possibly originated from the kidney according to immunohistochemical staining. In this rare case of fever of unknown origin, the primary lesion was a renal tumor with bone, liver, and brain metastases. Enhanced CT and PET/CT provided negative results, and the diagnosis was eventually confirmed by repeated bone marrow pathology.

A 65-year-old male was admitted to Peking Union Medical College Hospital. The patient had intermittent fever for 2 months with a maximum body temperature of 39.3 ℃ and elevated serum creatinine levels for 1 week. He had no other suggestive symptoms or positive signs. Laboratory test results suggested acute kidney injury and a sharp elevation in serum lactic dehydrogenase levels. Abdominal enhanced computed tomography (CT) revealed multiple low-density lesions, and further biopsy pathology demonstrated chronic inflammation. Thereafter, positron emission tomography (PET)/CT showed abnormally elevated uptake value for the bones throughout the entire body, in addition to the liver and brain. Repeated bone marrow biopsy finally confirmed metastatic bone cancer, which possibly originated from the kidney according to immunohistochemical staining. In this rare case of fever of unknown origin, the primary lesion was a renal tumor with bone, liver, and brain metastases. Enhanced CT and PET/CT provided negative results, and the diagnosis was eventually confirmed by repeated bone marrow pathology.

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.

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*

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*

Objective:To investigate the impact of sexual abstinence (SA) on semen parameters in oligospermia and asthenozoospermia.Methods:This retrospective cohort study was conducted for the clinical data from oligospermia ( n=5127), asthenozoospermia ( n=4003) and normozoospermia ( n=4529) with different SAs (2 d,3 d,4 d,5 d,6 d,7 d) who were treated in Andrology Department of West China Second University Hospital during January 2018 to December 201 9. The correlation between semen parameters (sperm concentration, motility, total motile sperm count etc.) and SA were observed. Results:In normozoospermia, as the SA was getting longer, the semen volume was increased from (3.3±1.2) mL to (4.1±1.3) mL ( r=0.167, P<0.001), the sperm concentration was increased from (89.0±60.9)×10 6/mL to (125.2±82.3)×10 6/mL ( r=0.181, P<0.001), the total sperm count was increased from (273.2±169.8)×10 6/ejaculate to (473.5±193.7)×10 6/ejaculate ( r=0.310, P<0.001). The progressive motility was decreased from 62.1%±13.0% to 59.5%±13.3% ( r=-0.057, P<0.001). The viability was decreased from 80.6%±8.5% to 79.0%±9.1% ( r=-0.048, P<0.001). In oligospermia, the semen volume was increased from (3.1±1.4) mL to (3.9±1.6) mL ( r=0.171, P<0.001) and the sperm concentration was decreased from (10.3±5.5)×10 6/mL to (8.7±4.3)×10 6/mL ( r=-0.043, P<0.001), the total sperm count was increased from (29.0±17.1)×10 6/ejaculate to (38.6±19.8)×10 6/ejaculate ( r=0.285, P<0.001). The progressive motility was decreased from 41.1%±17.0% to 35.1%±17.3% ( r=-0.141, P<0.001). The viability was decreased from 71.1%±12.3% to 63.1%±16.6% ( r=-0.225, P<0.001). The peak value of sperm concentration and motility were observed on day 2. In asthenozoospermia, the semen volume was increased from (3.1±1.4) mL to (3.8±1.9) mL ( r=0.197, P<0.001) and the sperm concentration was increased from (35.1±30.5)×10 6/mL to (49.7±31.9)×10 6/mL ( r=0.071, P<0.001), the total sperm count was increased from (109.1±82.3)×10 6/ejaculate to (170.1±99.3)×10 6/ejaculate ( r=0.394, P<0.001). The viability was decreased from 59.6%±16.4% to 54.0%±16.4% ( r=-0.081, P<0.001). The SA was not related to motility and round cells ( P>0.05). TMSC was slightly increased with extending EAT ( r=0.119, P<0.001). Conclusion:The longer SA has positive influence on semen volume and total sperm count on different levels in males with normozoospermia, oligospermia and asthenozoospermia. The viability was decreased by extended SA. Shortening the SA was likely to help oligospermic patients conduct sperm with higher concentration, motility, viability and normal morphology. TMSC was not significantly increased by extended SA. Asthenozoospermia can conduct sperm with higher concentration and TMSC when applying longer SA. The progressive motility was not significantly influenced by SA.

Objective:To investigate the impact of sexual abstinence (SA) on semen parameters in oligospermia and asthenozoospermia.Methods:This retrospective cohort study was conducted for the clinical data from oligospermia ( n=5127), asthenozoospermia ( n=4003) and normozoospermia ( n=4529) with different SAs (2 d,3 d,4 d,5 d,6 d,7 d) who were treated in Andrology Department of West China Second University Hospital during January 2018 to December 201 9. The correlation between semen parameters (sperm concentration, motility, total motile sperm count etc.) and SA were observed. Results:In normozoospermia, as the SA was getting longer, the semen volume was increased from (3.3±1.2) mL to (4.1±1.3) mL ( r=0.167, P<0.001), the sperm concentration was increased from (89.0±60.9)×10 6/mL to (125.2±82.3)×10 6/mL ( r=0.181, P<0.001), the total sperm count was increased from (273.2±169.8)×10 6/ejaculate to (473.5±193.7)×10 6/ejaculate ( r=0.310, P<0.001). The progressive motility was decreased from 62.1%±13.0% to 59.5%±13.3% ( r=-0.057, P<0.001). The viability was decreased from 80.6%±8.5% to 79.0%±9.1% ( r=-0.048, P<0.001). In oligospermia, the semen volume was increased from (3.1±1.4) mL to (3.9±1.6) mL ( r=0.171, P<0.001) and the sperm concentration was decreased from (10.3±5.5)×10 6/mL to (8.7±4.3)×10 6/mL ( r=-0.043, P<0.001), the total sperm count was increased from (29.0±17.1)×10 6/ejaculate to (38.6±19.8)×10 6/ejaculate ( r=0.285, P<0.001). The progressive motility was decreased from 41.1%±17.0% to 35.1%±17.3% ( r=-0.141, P<0.001). The viability was decreased from 71.1%±12.3% to 63.1%±16.6% ( r=-0.225, P<0.001). The peak value of sperm concentration and motility were observed on day 2. In asthenozoospermia, the semen volume was increased from (3.1±1.4) mL to (3.8±1.9) mL ( r=0.197, P<0.001) and the sperm concentration was increased from (35.1±30.5)×10 6/mL to (49.7±31.9)×10 6/mL ( r=0.071, P<0.001), the total sperm count was increased from (109.1±82.3)×10 6/ejaculate to (170.1±99.3)×10 6/ejaculate ( r=0.394, P<0.001). The viability was decreased from 59.6%±16.4% to 54.0%±16.4% ( r=-0.081, P<0.001). The SA was not related to motility and round cells ( P>0.05). TMSC was slightly increased with extending EAT ( r=0.119, P<0.001). Conclusion:The longer SA has positive influence on semen volume and total sperm count on different levels in males with normozoospermia, oligospermia and asthenozoospermia. The viability was decreased by extended SA. Shortening the SA was likely to help oligospermic patients conduct sperm with higher concentration, motility, viability and normal morphology. TMSC was not significantly increased by extended SA. Asthenozoospermia can conduct sperm with higher concentration and TMSC when applying longer SA. The progressive motility was not significantly influenced by SA.

Globozoospermia is a rare (0.1%) but severe disorder in male infertility, with round-headed and acrosomeless sperm defects. The pathogenesis of globozoospermia most probably originates in spermiogenesis, more specifically in acrosome formation and sperm head elongation. It is confirmed with genetic origin. Five human genes and 17 mouse genes have been so far confirmed in presenting globozoospermia. Intracytoplasmic sperm injection (ICSI) combined with assisted oocyte activation (AOA) has been proved to be a very efficient fertilization technique when compared with conventional ICSI in globozoospermia. In this paper, the discovery, recognition and definition of round head sperm, morphological characteristics and ultrastructure, related genes and pathogenesis, and the outcome of assisted reproduction in globozoospermic patients were summarized and discussed.

Globozoospermia is a rare (0.1%) but severe disorder in male infertility, with round-headed and acrosomeless sperm defects. The pathogenesis of globozoospermia most probably originates in spermiogenesis, more specifically in acrosome formation and sperm head elongation. It is confirmed with genetic origin. Five human genes and 17 mouse genes have been so far confirmed in presenting globozoospermia. Intracytoplasmic sperm injection (ICSI) combined with assisted oocyte activation (AOA) has been proved to be a very efficient fertilization technique when compared with conventional ICSI in globozoospermia. In this paper, the discovery, recognition and definition of round head sperm, morphological characteristics and ultrastructure, related genes and pathogenesis, and the outcome of assisted reproduction in globozoospermic patients were summarized and discussed.