Objective To find out the differences in gene expression of spleen tissue in septic rats by using DNA microarrays.Methods Thirty male Wistar rats were randomly (random number) and equally divided into control group and sepsis group,and septic rat model was induced by cecal ligation puncture (CLP).The rats of control group were only subjected to a simulated operation without CLP.Gene expression profiles were studied by using RatRef-12 gene chip.Rat gene expression profile was showed by using microarray to detect the changes in gene expression pattern of rat spleen tissue after CLP.And subsequently,by using relevant computer software to screen and analyze,the comparison of differences in gene expression between the sepsis group and control group was made.Results Of 22 523 genes,205 differential genes were found between sepsis group and control group,accounting for 0.910％.Among them 98 genes showed up-regulation,with 48 known functional genes,and 107 genes showed down-regulation,with 64 known functional genes.The function of such different genes were associated mainly with apoptosis,inflammation and energy metabolism of spleen cells.Conclusions Splenic dysfunction may be attibuted to the abnormal expression of relevant genes subjected to apoptosis,inflammation and alteration of energy metabolism.It may be the cause of immunosuppression in the later stage of sepsis.

Objective To explore the effect of UTI (Ulinastatin) preconditioning on gene expression profiles of spleen tissue in septic rats by DNA microarray technology. Methods Forty-five male Wistar rats were equally divided into sham group,sepsis group and UTI group by means of random number table.In UTI group the rats were treated with intramuscular injection of UTI( 105 U/kg) one hour before cecal ligation and puncture.In sepsis group and sham group intramuscular balanced solution (5 ml/kg) was given.Cecal ligation and puncture was used to reproduce septic rat model. Gene expression profile was studied by using RatRef-12 Rat gene expression profile microarray to detect the changes in gene expression pattern of rat spleen tissue after cecal ligation and puncture.Then using related computer software was used to screen and analyze the relationship between the Sepsis/UTI group and sham group. Results In 22 523 genes,205 differential genes were found between sepsis group and sham group,accounting for 0.910％.Among them 98 genes were up-regulated,with 48 known functional genes and 32 genes only showed in this group;107 genes were down-regulated,with 64 known functional genes and 34 genes only showed in it.197 differential genes were found in UTI group and sham group,accounting for 0.875％.Among them 114 genes were up-regulated,with 35 known functional genes and 19 genes only showed in this group; 83 genes were down-regulated,with 49 known functional genes and 19 genes only showed in it. Conclusions Abnormal expression of genes in the spleen tissue of rats with sepsis owing to excessive inflammation and immune suppression were partly relieved by UTI preconditioning.UTI pretects spleen at genetic level.

Objective:To explore the molecular mechanism for bone mass loss caused by staphylococcus aureus infection.Methods:Thirty 8-week-old male C57BL/6 mice were randomly divided into 3 groups ( n=10): control, infection and infection+JAK inhibitor (JAKi) ones. The mice were killed 2 weeks later for sampling from the femur and tibia. Micro-CT reconstruction was performed for analyses of BV/TV, Tb.N, Tb.Th and Tb.Sp to detect changes in bone mass; OCN immunohistochemistry and Goldner's trichrome staining were used to quantify osteoblasts; TRAP staining was used to quantify osteoclasts; the GSE166522 data set was downloaded and analyzed to explore the relationships between staphylococcus aureus infection and bone cell senescence and JAK/STAT pathway. Senescence β-Galactosidase staining, Osterix and P16 immunofluorescence colocalization were used to observe the changes in number of senescent cells. Results:MicroCT results showed a statistically significant difference in the loss of cancellous bone in the target area in the infection group compared with the control group ( P<0.05). The results of osteocalcin immunohistochemistry and Goldner's trichrome staining indicated that the number of osteoblasts in the infection group was significantly reduced ( P<0.05). TRAP staining indicated no significant difference in the number of osteoclasts between the infection and control groups ( P>0.05). Bioinformatics analysis found that staphylococcus aureus infection caused bone cell senescence and the JAK/STAT pathway was activated after the infection. Senescence β-Galactosidase staining suggested that senescent cells increased in the infection group compared with the control group. The number of Osterix and P16 positive senescent osteoprogenitor cells in the infection group was increased significantly compared with the control group. The number of senescent osteoprogenitor cells in the infection+JAKi group was significantly reduced and the bone loss was partially reversed after treatment of JAK inhibitor, compared with the infection group. Conclusion:Staphylococcus aureus may induce osteoprogenitor cell senescence through the JAK/STAT pathway and eventually lead to bone mass loss.

OBJECTIVETo detect potential mutation of EXT1 gene in a pedigree affected with multiple osteochondroma and explore its pathogenic mechanism.

METHODSThe coding regions and their flanking sequences of the EXT1/EXT2 genes were subjected to PCR amplification and Sanger sequencing. Suspected mutations were verified by excluding possible single nucleotide polymorphisms and bioinformatics analysis. Transcripts of the EXT1 gene in the proband were analyzed by TA clone-sequencing, with its abundance compared with that of healthy controls.

RESULTSDNA sequencing has identified in the proband a novel heterozygous point mutation (c.1164+1G to A) at the 5'splice sites of intron 3 of the EXT1 gene. The same mutation was not found in the healthy controls. Bioinformatics analysis indicated that the mutation is highly conserved and can lead to skipping of exon 3 or aberrant splicing. TA clone-sequencing indicated that the numbers of transcripts with skipping of exon 3 has significantly increased in the proband (< 0.05) compared with the controls.

CONCLUSIONThe c.1164+1G to A mutation has resulted in skipping of exon 3 in a proportion of EXT1 gene transcripts. As the result, the number of transcripts with tumor suppressing function is relatively reduced and has ultimately led to the tumors.

Adult ; Base Sequence ; Child ; Exostoses, Multiple Hereditary ; genetics ; Female ; Humans ; Male ; Molecular Sequence Data ; N-Acetylglucosaminyltransferases ; genetics ; Point Mutation ; RNA Splice Sites ; RNA Splicing

OBJECTIVE: To explore the genetic basis for a pedigree affected with hereditary multiple osteochondroma (HMO). METHODS: Peripheral blood samples were collected from the proband and members of his pedigree with informed consent. Following extraction of genomic DNA, all coding exons and flanking intronic sequences (-10 bp) of the EXT1 and EXT2 genes were subjected to targeted capture and next generation sequencing (NGS). Suspected variant was verified by Sanger sequencing. RESULTS: A heterozygous nonsense variant (c.1911C>A) was found in exon 10 of the EXT1 gene in the proband and his affected father but not in a healthy sister and normal controls. The variant was classified as a pathogenic based on the guidelines of the American College of Medical Genetics and Genomics (PVS1+PM2+PP1). Bioinformatic analysis predicted that the c.1911C>A variant may be disease-causing via nonsense-mediated mRNA decay and anomalous splicing. CONCLUSION The c.1911C>A variant probably underlay the disease in this pedigree. Discovery of this variant enriched the variant spectrum of HMO.
Codon, Nonsense ; Exons/genetics* ; Exostoses, Multiple Hereditary/genetics* ; Heterozygote ; Humans ; Pedigree

Objective:To establish the urodynamic classification of middle-aged and elderly men with benign prostatic obstruction(BPO), and to analyze the efficacy of transurethral resection of the prostate(TURP) on various types of patients.Methods:A retrospective analysis of middle-aged and elderly male patients with non-neurogenic lower urinary tract symptoms(LUTS) who underwent urodynamic tests from January 2010 to December 2018, including 793 patients with BPO. Urodynamics examination of detrusor without contraction needs to complete cystoscopy to diagnose BPO. During urodynamic examination, the detrusor uninhibited contraction induced by spontaneous or stimulation during the bladder filling period is diagnosed as overactivity of the bladder detrusor(DO), and the LinPURR chart indicates the detrusor underactivity(DU). Based on the persistence of BPO leading to DO, DU, and decreased bladder compliance, 793 male patients with BPO with LUTS were divided into four types, including type Ⅰ(BPO: n=164, 20.7%), type Ⅱ(BPO combined with DO: n=333, 42.00%), type Ⅲ(BPO combined with DU: n=267, 33.7%), type Ⅳ(BPO combined with decreased bladder compliance: n=29, 3.7%). The preoperative comparison between groups showed that the age of type Ⅰ-Ⅳ gradually increased, and the age of type Ⅰ was significantly smaller than other types [(67.3±8.2)years, (69.7±7.7)years, (71.5±7.9)years, (72.4±7.1)years, P<0.05]. Compared with other types, the type Ⅰ’s IPSS-S[(9.1±3.6)points vs.(10.4±3.1) points, (9.2±3.3) points, (10.4±3.1)points, P<0.05], IPSS-V[(13.5±3.4) points vs. (14.2±3.5)points, (14.0±3.5)points, (14.2±2.9)points, P<0.05], IPSS scores[(22.6±5.4)points, (24.7±4.9)points, (23.1±5.3)points, (24.6±4.7)points, P<0.05] were significantly lower than other groups, the maximum bladder capacity [(332.6±83.2)ml vs.(221.4±80.8)ml, (286.7±108.2)ml, (242.3±103.4)ml, P<0.05], the functional bladder capacity was significantly higher than other types[(215.2±90.0)ml, (148.5±76.0)ml, (154.9±87.2)ml, (121.2±72.9)ml, P<0.05]. Type Ⅱ’s IPSS-S[(10.4±3.1)points vs.(9.1±3.6)points, (9.2±3.3)points, P<0.05], nocturia frequency[(3.7±1.8)times vs.(3.2±1.8)times, (3.2±1.6)times, P<0.05], IPSS score[(24.7±4.9)points vs.(22.6±5.4)points, (23.1±5.3)points, P<0.05], quality of life scores [(4.9±0.9) points, (4.6±0.9)points, (4.6±0.9)points, P<0.05] was significantly higher than type Ⅰ and type Ⅲ ( P<0.05). Type Ⅲ and Ⅳ had higher residual urine than type Ⅱ[(121.3±96.4)ml, (121.3±96.4)ml vs.(71.2±73.5)ml, P<0.05]. Type Ⅳ’s IPSS-S[(10.4±3.1)points vs. (9.1±3.6)points, (9.2±3.3)points, P<0.05], IPSS-V[(14.2±2.9) points vs.(13.5±3.4)points, (14.0±3.5)points, P<0.05], the frequency of nocturia[(3.8±1.9)times vs.(3.2±1.8)times, (3.2±1.6)times, P<0.05] was significantly higher than that of type Ⅰ and type Ⅲ, and the quality of life score was higher than type Ⅰ and type Ⅲ[(4.3±0.8)points vs.(4.7±0.9)points, (4.6±0.9)points, P<0.05]. type Ⅱ and type Ⅳ’s bladder compliance[(21.4±24.2)ml/cmH 2O, (11.0±11.4)ml/cmH 2O vs.(33.9±23.7)ml/cmH 2O, (33.1±32.7)ml/cmH 2O, P<0.05], maximum bladder capacity[(221.4±80.8)ml, (242.3±103.4)ml vs.(332.6±83.2)ml, (286.7±108.2)ml, P<0.05], functional bladder capacity[(148.5±76.0)ml, (121.2±72.9)ml vs.(215.2±90.0)ml, (154.9±87.2)ml, P<0.05] were significantly less than type Ⅰ and type Ⅲ( P<0.05). From November 2016 to November 2018, 60 middle-aged and elderly male patients with confirmed BPO and TURP were selected, including type Ⅰ( n=17, 28.3%), type Ⅱ ( n=23, 38.3%), and Ⅲ type ( n=11, 18.3%), Ⅳ type( n=9, 15.1%). Type IV patients are significantly older than other types ( P<0.05), bladder compliance is significantly worse than other types( P<0.05), the maximum bladder capacity is smaller than other types( P<0.05). The follow-up started 3 months after the operation. The content of the follow-up included IPSS, IPSS-S, IPSS-V, nocturia frequency, undisturbed sleep time, nocturia quality of life score, and life quality score. Results:The IPSS scores of type Ⅰ, type Ⅱ, and type Ⅲ after TURP were significantly improved compared with preoperative(19.8±6.2 vs.3.4±1.8; 21.9±5.2 vs.4.6±2.6; 21.5±6.2 vs.5.7±4.6, P<0.05), type Ⅳ urine storage symptom score (9.1±4.1 vs.4.3±3.7), nocturia frequency(3.6±1.5vs.2.3±1.6), nocturia quality of life score (25.3±6.9 vs.31.4±13.7) Compared with preoperatively, there was no significant improvement( P>0.05). The quality of life score improvement of type Ⅳ patients was significantly lower than that of type Ⅰ, type Ⅱ, and type Ⅲ (10.9±9.1 vs.12.2±9.0, 14.4±5.7, 12.7±5.8, P<0.05). The IPSS score of type Ⅳ patients was significantly higher than that of type Ⅰ(7.0±5.8 vs.3.4±1.8), and the nocturia quality of life score was significantly lower than that of each group (31.4±13.7 vs.37.5±4.2, 38.7±3.5, 37.8±3.8, P<0.05). Conclusions:For middle-aged and elderly men with BPO, we divide them into four types based on the results of urodynamic examinations, type Ⅰ(simple BPO), type Ⅱ(BPO combined with DO), type Ⅲ(BPO combined with DU), type Ⅳ(BPO combined with bladder compliance decline). Type Ⅰ patients have the best bladder function, and TURP has the best effect; type Ⅱ has a high symptom score and poor quality of life, and can benefit after TURP; type Ⅲ bladder function is poor, and surgery should be performed as soon as possible to prevent further deterioration of bladder function; type Ⅳ bladder function is the best poor, IPSS score and quality of life score are high, TURP surgery is not effective.

Objective:To explore the mechanism of Xijiao Dihuang Ddecoction (XJDHT) against sepsis-induced liver injury based on transcriptomics.Methods:Sixty C57BL/6 mice were randomly (random number) divided into the sepsis group, sepsis treatment with XJDHT and control group, with 20 mice in each group. The sepsis mouse model was established by intraperitoneal (i.p.) injection of lipopolysaccharide (LPS). The control group was intraperitoneally injected with the same amount of normal saline. The sepsis treatment with XJDHT group was injected with XJDHT (crude drug 187.5 mg) twice a day 2 days before modeling. After modeling, gastric feeding was continued twice a day, while the control group and sepsis group were gavaged with the same amount of normal saline. At 72 h after LPS intervention, 9 mice in each group were randomly selected. After anesthesia, part of the liver were taken for small RNA and RNA sequencing and analysis, and part of the liver were taken for pathological examination.Results:XJDHT could improve the histopathological changes of liver in septic mice, and alleviate some abnormally expressed microRNAs (mmu-mir-292a-5p, mmu-mir-871-3p, mmu-mir-653-5p, mmu-mir-293-5p, mmu-mir-155-3p, mmu-mir-346-5p, mmu-mir-187-5p, mmu-mir-3090-3p) and their target genes.Conclusions:XJDHT can reduce the liver histopathological changes in septic mice, and its mechanism may be related to XJDHT regulating the expression of important key genes of liver of sepsis like mmu-mir-187-5p and its target genes such as ADAM8, irak3 and PFKFB3

【Objective】 To explore the surgical treatments and therapeutic outcomes for benign testicular tumor. 【Methods】 Clinical data of 53 patients with benign testicular tumor treated with surgery during May 2004 and Jul.2021 were retrospectively analyzed. 【Results】 The postoperative pathological diagnosis of 53 patients included 33 patients with epidermal cysts, 12 with mature teratomas, 2 with bilateral testicular tumors (one of them was epidermal cysts in the left and mature teratoma in the right, and the other was bilateral leiomyomas), and 6 benign cases. Testis sparing surgery (TSS) group had 23 patients and radical orchiectomy (RO) group had 30 patients. There were no significant differences in patients’ age, tumor location, disease course, and ultrasound examination results between the two groups (P>0.05). The tumor size of the RO group was (2.60±0.94) cm, which was larger than that of the TSS group (1.55±0.52) cm (P<0.001). There were no statistically significant differences in surgical time and postoperative hospital stay between the two groups (P>0.05). A total of 15 patients (13 with TSS and 2 with RO) underwent intraoperative frozen rapid pathological examination (FSA), which was consistent with post-operative paraffin pathological results. Durign the follow up of 2-219 months,median 38 months, there was no recurrence in either groups. 【Conclusion】 Testis sparing surgery is a reliable treatment modality for benign testicular tumor, which may also decrease the level of androgen and incidence of asthenozoospermia. It can be considered for tumors less than 2 cm with benign tendency or uncertain nature.