[Abstract] Objective To study the morphology, muscle architecture index and distribution pattern of intramuscular nerve dense area of elbow muscle, so as to provide anatomical location for poster-lateral approach of elbow joint and transplantation of elbow muscle flap. Methods Through gross anatomy, muscle architecture index and modified Sihler’s intramuscular nerve staining, 10 cases with an average age of 64. 2 years were selected. Results The elbow muscle was approximate triangle, the muscle wet weight was (6. 31±0. 85) g, the muscle length was (6. 24±0. 78) cm, the muscle fiber length was (4. 74±0. 88) cm, pennation angle(70. 60±6. 41)°and the muscle physiological cross-sectional area was (0. 41±0. 15) cm

OBJECTIVE: The leukemia cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model. METHODS: Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated, and the leukemia cells were inoculated into NCG mice via tail vein. The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry, and the infiltration of leukemia cells in bone marrow, liver, spleen and other organs of the mice was detected by pathology and immunohistochemistry. After the first generation mice model was successfully established, the spleen cells from the first generation mice were inoculated into the second generation mice, and after the second generation mice model was successfully established, the spleen cells from the second generation mice were further inoculated into the third generation mice, and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model. RESULTS: On the 10th day after inoculation, hCD45⁺ leukemia cells could be successfully detected in the peripheral blood of the first generation mice, and the proportion of these cells was gradually increased. On average, the mice appeared listless 6 or 7 weeks after inoculation, and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice. The spleen of the mice was obviously enlarged, and immunohistochemical examination showed that hCD3⁺ leukemia cells infiltrated into bone marrow, liver and spleen extensively. The second and third generation mice could stably develop leukemia, and the average survival time was 4-5 weeks. CONCLUSION Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts (PDTX) model.
Humans ; Animals ; Mice ; Precursor T-Cell Lymphoblastic Leukemia-Lymphoma ; Heterografts ; Bone Marrow ; Disease Models, Animal ; T-Lymphocytes ; Mice, SCID

OBJECTIVE: To investigate the inflammatory effects of Cinobufotalin on monocytes in resting state and macrophages in activated state and its molecular mechanism. METHODS: THP-1 cells were stimulated with Phorbol 12-myristate 13-acetate to induce differentiation into macrophages. Lipopolysaccharides was added to activate macrophages in order to establish macrophage activation model. Cinobufotalin was added to the inflammatory cell model for 24 h as a treatment. CCK-8 was used to detect cell proliferation, Annexin V /PI double staining flow cytometry was used to detect cell apoptosis, flow cytometry was used to detect macrophage activation, and cytometric bead array was used to detect cytokines. Transcriptome sequencing was used to explore the gene expression profile regulated by Cinobufotalin. Changes in the significantly regulated molecules were verified by real-time quantitative polymerase chain reaction and Western blot. RESULTS: 1∶25 concentration of Cinobufotalin significantly inhibited the proliferation of resting monocytes(P<0.01), and induced apoptosis(P<0.01), especially the activated macrophages(P<0.001, P<0.001). Cinobufotalin significantly inhibited the activation of macrophages, and significantly down-regulated the inflammatory cytokines(IL-6, TNF-α, IL-1β, IL-8) released by activated macrophages(P＜0.001). Its mechanism was achieved by inhibiting TLR4/MYD88/P-IκBa signaling pathway. CONCLUSION Cinobufotalin can inhibit the inflammatory factors produced by the over-activation of macrophages through TLR4/MYD88/P-IκBa pathway, which is expected to be applied to the treatment and research of diseases related to the over-release of inflammatory factors.
Humans ; Toll-Like Receptor 4/metabolism* ; Myeloid Differentiation Factor 88/genetics* ; Macrophages/metabolism* ; Cytokines/metabolism* ; Lipopolysaccharides/pharmacology* ; NF-kappa B

Aim To compare the effects of BoneCeramic and Bio-Oss on the proliferation, adhesion and osteogenic differentiation of dog bone marrow mesenchymal stem cells (DBMSCs) in vitro. Methods DBM-SCs were isolated and cultured in vitro and identified for osteogenic and adipogenic differentiation. CCK-8 was used to detect the effects of BoneCeramic and Bio-Oss bone meal extracts on the vitality of DBMSCs. SEM was used to observe the adhesion of DBMSCs on the surface of bone meal. ALP was used to quantitatively detect the osteogenic differentiation of DBMSCs on the surface of bone meal, and the relative expressions of OPN, OCN, RUNX-2, and ALP genes were detected by RT-PCR. Results The cells isolated and cultured from dog bone marrow conformed to the characteristics of mesenchymal stem cells. CCK-8 results showed that the stock solutions of Bio-Oss and Bone Ceramic had no obvious cytotoxic effect on DBMSCs. SEM showed that DBMSCs were distributed more widely on the surface of BoneCeramic than that of Bio-Oss. ALP quantitative detection showed that DBMSCs on the BoneCeramic surface were more conducive to its osteogenic differentiation than Bio-Oss, and it also increased the expression of OPN, OCN, RUNX-2 and ALP genes. Conclusions In vitro studies show that both Bio-Oss and BoneCeramic have good biocompatibility. Compared with Bio-Oss bone meal, BoneCeramic is more conducive to the osteogenic differentiation of DBMSCs.

Objective:To observe the effect of acupuncture on the expression of mitochondrial proteome in hippocampus of senescence-accelerated mouse prone g (SAMPg) mice models with Alzheimer disease (AD),and to explore the possible protective mechanism of acupuncture on mitochondria.Methods:Sixty 6-month-old male SAMP8 mice were randomly divided into an acupuncture at acupoint group,an acupuncture at non-acupoint group and a model group,20 mice in each group.The 20 male senescence-accelerated mouse/resistance 1 (SAMR1) mice of the same age were used as a normal control group.Shenshu (BL 23),Baihui (GV 20),Xuehai (SP 10) and Geshu (BL 17) were selected for acupuncture intervention in acupuncture at acupoint group.After an 8-week intervention,mitochondrial tissues were extracted from the hippocampus.Differentially expressed proteins were identified by subcellular organelle proteomics.Western blot was used to verify the expressions of some related proteins in hippocampal mitochondria.Results:Compared with the model group,there were 13 differentially expressed protein spots in the acupuncture at acupoint group,of which,9 were up-regulated,including neurofilament light polypeptide (NFL),actin (cytoplasmic 1,database ID:ACTB),tubulin beta-2A chain (TBB2A),tropomodulin-2 (TMOD2),pyruvate dehydrogenase E1 component subunit beta (PDHE1-β),NADH-ubiquinone oxidoreductase 75 kDa subunit (database ID:NDUS1),heat shock cognate 71 kDa protein (HSC71),pyruvate dehydrogenase E1 component subunit alpha (PDHE1-α) and ATP synthase beta subunit (ATP-β);4 were down-regulated,including glial fibrillary acidic protein (GFAP),pyruvate dehydrogenase phosphatase 1 (PDP1),mitochondrial-processing peptidase subunit alpha (MMP-α) and adenosine kinase (ADK).According to the information provided in the protein database,most of the differentially expressed proteins involve the regulation of mitochondrial function and structure.The expression levels of NFL and TBB2A in the normal control group and the acupuncture at acupoint group were significantly higher than those in the acupuncture at non-acupoint group (P＜0.05).ATP-β and NDUS1 expression levels were significantly higher in the acupuncture at acupoint group than those in the acupuncture at non-acupoint group (P＜0.05);there was no significant difference between the acupuncture at non-acupoint group and the model group (P＞0.05).Conclusion:Acupuncture may achieve the potential therapeutic effect on AD by regulating the structure and functional proteins of hippocampal mitochondria.

Objective To evaluate the clinical efficacy of drospirenone and ethinylestradiol tablets combined with gonadotropin-releasing hormone agonistin(GnRH-a) in treatment of patients with uterine muscular glands and its effect on serum ovarian cancer antigen (CA125) and human epididymis protein (HE4).Methods A total of 96 patients with adenomyosis were randomly divided into control group and treatment group,each group 48 cases.Control group was given levonorgestrel intrauterine birth control system,placed levonorgestrel intrauterine system,given drospirenone ethinyl estradiol tablets (ethinyl estradiol 0.03 mg and drospirenone 3 mg),once a day,orally given for 21 d,and stopped for 7 d,then continue for 3 months after discontinuation of oral,according to this method for 3 consecutive months medication.Treatment group was given GnRH-a 3.75 mg every 4 weeks a time before treatment,subcutaneous injection for 3 times.The clinical effect,cancer antigen-125 (CA125),human epididymis protein 4 (HE4),B lymphocyte tumor-2 (Bcl-2),Bcl-2 relative X protein (Bax),caspase-3 and adverse drug reactions in two groups were observed.Results After treatment,the clinical efficacy in control group was 81.25％ (39/48 cases),had significant difference with that in treatment group,which was 95.83％ (46/48 cases,P ＜0.05).The levels of serum CA125,HE4 and Bcl-2 and uterine volume in treatment group were (27.22 ±0.35) U · L-1,(20.67 ±0.52) pmol · L-1,(3.62 ±0.43) μg · mL-1,(95.11 ± 10.29)cm2,had significant difference with those in control group,which were (41.42 ± 0.43) U · L-1,(36.67 ± 0.38) pmol · L-1,(4.76 ± 0.52) μg · mL-1,(120.02 ± 13.92) cm2 (P ＜0.05).The levels of Bax and caspase-3 in treatment group had significant difference with control group (P ＜ 0.05).The adverse drug reactions in treatment group were agitation,nausea,intermenstrual bleeding,total incidence rate of adverse drug reactions was 6.25％ (3/48 cases).The adverse drug reactions in control group were nausea,intermenstrual bleeding,skin rash,total incidence rate of adverse drug reactions was 14.58％ (7/48 cases,P ＞ 0.05).Conclusion Drospirenone and ethinylestradiol tablets in the treatment of adenomyosis was effective with high safety.

OBJECTIVETo investigate the risk factors for the development of congenital anal atresia in neonates.

METHODSA total of 70 neonates who were admitted to 17 hospitals in Foshan, China from January 2011 to December 2014 were enrolled as case group, and another 70 neonates who were hospitalized during the same period and had no anal atresia or other severe deformities were enrolled as control group. Univariate and multivariate logistic regression analyses were used to investigate the risk factors for the development of congenital anal atresia.

RESULTSThe univariate analysis revealed that the age of mothers, presence of oral administration of folic acid, infection during early pregnancy, and polyhydramnios, and sex of neonates showed significant differences between the case and control groups (P<0.05). The multivariate logistic regression analysis revealed that infection during early pregnancy (OR=18.776) and male neonates (OR=9.304) were risk factors for congenital anal atresia, and oral administration of folic acid during early pregnancy was the protective factor (OR=0.086).

CONCLUSIONSInfection during early pregnancy is the risk factor for congenital anal atresia, and male neonates are more likely to develop congenital anal atresia than female neonates. Supplementation of folic acid during early pregnancy can reduce the risk of congenital anal atresia.

Anus, Imperforate ; etiology ; Female ; Humans ; Infant, Newborn ; Logistic Models ; Male ; Pregnancy ; Risk Factors

Objective To investigate mutational spectrum and frequency of the mitochondrial 12S rRNA gene in Chinese subjects with aminoglycoside-induced and non-syndromic hearing loss.Methods Total of 456 subjects with non-syndromic hearing loss were recruited from seven schools for deaf-mutes in Zhejiang province.Genomic DNA was extracted from the whole blood,and then the DNA fragment was amplified spanning the 12S rRNA gene,followed by sequencing and analyzed.Results Thirty-one variants were identified by mutation analysis of 12S rRNA gene in these subjects.The frequency of the known 1555A ＞ G mutation was 4.4％ (20/456).Prevalence of other putative deafness-associated mutation at positions 961 and 1095 were 2.0％ (9/456) and 0.7％ (3/456) respectively.Furthermore,the 1027A ＞ G,1109T ＞ C and 1431G＞A variants conferred increased sensitivity to ototoxic drugs or non-syndromic deafness as they were absent in 449 Chinese controls and localized at highly conserved nucleotides of this 12S rRNA gene.Moreover,clinical data showed a wide range of age-of-onset,variety of severity and various audiometric configurations in subjects carrying the 1555A＞G mutation.Conclusions Our data demonstrated that the mitochondrial 12S rRNA gene is the hot spot for mutations associated with aminoglycoside ototoxicity and non-syndromic hearing loss.Nuclear modifier genes,mitochondrial haplotypes and environmental factors might play a role in the phenotypic manifestation of these mutations.

WU polyomavirus (WUPyV), a new member of the genus Polyomavirus in the family Polyomaviridae, is recently found in patients with respiratory tract infections. In our study, the complete genome of the two WUPyV isolates (FZ18, FZTF) were sequenced and deposited in GenBank (accession nos. FJ890981, FJ890982). The two sequences of the WUPyV isolates in this study varied little from each other. Compared with other complete genome sequences of WUPyV in GenBank (strain B0, S1-S4, CLFF, accession nos. EF444549, EF444550, EF444551, EF444552, EF444553, EU296475 respectively), the sequence length in nucleotides is 5228bp, 1bp shorter than the known sequences. The deleted base pair was at nucleotide position 4536 in the non-coding region of large T antigen (LTAg). The genome of the WUPyV encoded for five proteins. They were three capsid proteins: VP2, VP1, VP3 and LTAg, small T antigen (STAg), respectively. To investigate whether these nucleotide sequences had any unique features, we compared the genome sequence of the 2 WUPyV isolates in Fuzhou, China to those documented in the GenBank database by using PHYLIP software version 3.65 and the neighbor-joining method. The 2 WUPyV strains in our study were clustered together. Strain FZTF was more closed to the reference strain B0 of Australian than strain FZ18.
Adult ; Child, Preschool ; China ; Evolution, Molecular ; Genome, Viral ; genetics ; Genomics ; Humans ; Male ; Molecular Sequence Data ; Phylogeny ; Polyomaviridae ; genetics ; isolation & purification ; Sequence Analysis, DNA ; methods

OBJECTIVETo study the clinical outcomes of stage I testis teratoma, including pure teratoma, and to provide information on the treatment options for this disease.

METHODSWe retrospectively analyzed 27 cases of orchiectomy for stage I testis teratoma, excluding epidermoid cyst, and investigated its recurrence associated with treatment methods and clinicopathological factors.

RESULTSFour of the 27 cases relapsed, all in the orchiectomy group and confined to the retroperitoneal region, 3 with and the other 1 without risk factors, but with no death. No recurrence was found in those treated by orchiectomy followed by chemotherapy with bleomycin, etoposide and platinum (BEP). The total rate of recurrence was 15.8%. No severe side effects were observed in the 9 patients undergoing adjuvant BEP chemotherapy.

CONCLUSIONRisk factors may increase the recurrence rate of stage I testis teratoma, while postoperative adjuvant chemotherapy can reduce it, including that of pure teratoma, though surveillance policy remains the most popular option after orchiectomy.

Adolescent ; Adult ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; pathology ; Neoplasm Staging ; Retrospective Studies ; Teratoma ; pathology ; therapy ; Testicular Neoplasms ; pathology ; therapy ; Young Adult