Objective To establish the background information of physiological parameters for germ?free ( GF ) Taihu piglets. Methods In this study we selected 25 days old GF Taihu piglets and 4 conventional ( CV) littermates, the male and female ratio was 1∶3, to measure the normal clinical values of hematology and serum biochemistry, immunoglobu?lin concentration and main organ coefficients. The analysis of relative growths of main organ weight to body weight was con?ducted in the Taihu GF and CV pigs by allometric scaling model. Results (1) Twelve hematological parameters and 8 blood biochemical parameters in the GF piglets were significantly lower than those in CV pigs (P<0?05). (2) The aver?age body weight, IgM concentration of GF pigs and CV pigs had significant difference ( P <0?05 ) , and no mesenteric lymph nodes were found in the GF pigs. (3) The gut weight had the largest allometric association with body weight in the GF pigs, while spleen weight has the largest allometric association with body weight in the CV pigs. Both the weight of heart and stomach in CV and GF pigs had a negative allometric association with body weight (allometric coefficient b<1), respectively. Conclusions Different microbe control grades affect the body weight, hematology and serum biochemistry, expression of immunoglobulin and development of main organs in laboratory pigs.

Objective:To explore the genetic etiology for a premature ovarian insufficiency (POI) patient from a consanguineous Chinese family, and to provide basis for genetic counseling and fertility counseling.Methods:Whole-exome sequencing was performed using DNA extracted from the blood sample of POI patient. Suspected pathogenic mutation was analyzed by bioinformatics methods and verified by Sanger sequencing. The pathogenicity of the variation was assessed according to the ACMG genetic variation classification criteria and guidelines.Results:A homozygous variation, c. 32G>T (p.G11V), of PSMC3IP was identified in the patient. Bioinformatics analysis revealed that the variation was conserved in different animal species, and this variation was classified as possible pathogenic variation according to the ACMG genetic variation classification criteria and guidelines.Conclusions:The homozygous missense variation of PSMC3IP is the cause of the POI patient in this family. We are reporting for the first time the missense variation in PSMC3IP gene caused POI, which enriched the mutation spectrum of PSMC3IP and provided the basis for genetic counseling and fertility guidance of this family.

OBJECTIVETo investigate the incidence of microsatellite instability (MSI) in sporadic colorectal carcinoma (CRC) using BAT-25 and BAT-26 loci, and its association with clinicopathological features.

METHODSMicrosatellite analysis was performed on tissue samples from 73 primary and 53 metastatic tumors collected at the Department of Pathology, Fudan University Cancer Hospital in 2002. Genomic DNA was extracted from paraffin-embedded tissues. Microsatellite alterations of BAT-25 and BAT-26 were detected using fluorescent PCR followed by fragment analysis on automatic DNA sequencer with GeneScan 3.1 software. A case of hereditary nonpolyposis colorectal cancer syndrome (HNPCC) with known high-frequency MSI (MSI-H) was included as positive control.

RESULTSEleven out of 73 samples from primary tumors (15.1%) were MSI-positive and significantly associated with patient age, tumor site, differentiation and prognosis (P < 0.05). There was no significant difference between the positive rate of MSI in tissue samples from primary and metastatic sites among the 53 metastatic tumors, being 17.0% and 13.2%, respectively, P > 0.05. Two cases with negative MSI at the primary site but positive at the metastatic sites were observed.

CONCLUSIONMSI is a frequent molecular event that may serve as a useful parameter for studying tumor biological behavior. MSI plays a partial role in the metastasis of sporadic CRC, but the role of mismatch repair genes and its exact mechanism remains to be determined. The classification of sporadic CRC according to MSI may be of importance both theoretically and practically.

Adenocarcinoma ; genetics ; secondary ; Adult ; Aged ; Colonic Neoplasms ; genetics ; pathology ; Colorectal Neoplasms, Hereditary Nonpolyposis ; genetics ; Female ; Follow-Up Studies ; Humans ; Liver Neoplasms ; genetics ; secondary ; Male ; Microsatellite Repeats ; Middle Aged ; Prognosis ; Rectal Neoplasms ; genetics ; pathology

The industry of germ-free animals has been a hot spot in research along with the rapid development of studies on the relationship between microbiota and host diseases. Because it is pathogen?free, and the high degree of simi?larity in anatomy, physiology, pathogenesis to humans, germ?free pig is considered a clinical relevant model to be widely used in life science research. Based on the current state of research of germ?free pig cultivation at home and abroad and the experimental studies carried out in our laboratory as well, this article gives a simple discussion on germ?free technique of domestic pigs.

OBJECTIVETo analyze the reasons of intraoperative complications of microendoscopic disectomy (MED) and corresponding preventive measures.

METHODSFrom October 2001 to January 2012, the data of 851 patients with lumbar disc herniation underwent MED were retrospectively analyzed. There were 469 males and 382 females with an average age of 42.5 years ranging 16 to 75. Course of disease was from 1 to 18 months with an average of 3 months. The segments of herniated disc including L3,4 of 24 cases, L4,5 of 418 cases and L5S1 of 409 cases . Main symptoms included low back pain with lower extremity radial pain and numbness. Of them,unilateral lower extremity symptom was in 729 cases and bilateral symptom was in 122 cases. There were at least 2 abnormal signs in the four signs which including feeling anormaly, muscle strength anormaly,dysreflexia and muscle atrophy. Distraction test of nerve was positive. CT or MRI findings must coincide with the clinical symptoms and signs. No lumbar instability,spinal stenosis,the upper lumbar disc herniation or combined with cauda equina nerve syndrome were found in 851 patients. The intraoperative complications were recorded and analyzed for the reasons of the intraoperative complication and related prevention measures.

RESULTSAccording to the Macnab standard,424 cases obstained excellent results, 321 good,106 fair,with excellent and good rate of 87.5%. The result was similar to the traditional open operation. One cases transferred to open operation due to equipment breakdown, case died for myocardial infarction at 11 days after the operation, 2 cases occurred acute epidural hematoma in 1 hour after operation. Injury of dura mate of spinal cord occurred in 28 cases and incidence rate was 3.29%(28/851); traction injury of nerve root occurred in 38 cases and incidence rate was 4.46% (38/851). One case occurred in retroperitoneal hematoma, 2 cases in incomplete cauda equina injury and 2 cases in incomplete nerve root breakage.

CONCLUSIONSkilled endoscopic hemostasis techniques,careful and meticulous operation is very important for the prevention of intraoperative complications. Moreover,timely finding and treating the complications was effective measures to prevent the coniplications.

Adolescent ; Adult ; Aged ; Diskectomy ; adverse effects ; Dura Mater ; injuries ; Endoscopy ; Female ; Hematoma, Epidural, Spinal ; etiology ; Humans ; Intraoperative Complications ; etiology ; prevention & control ; Male ; Middle Aged ; Retrospective Studies ; Spinal Nerve Roots ; injuries

Objective To study the clinical characteristics, pathological features, diagnosis,therapy and prognosis of primary small cell carcinoma of the larynx (PSCCL). Methods Six cases of PSCCL collected from 1990 to 2009 was retrospectively analyzed. The diagnosis was confirmed by pathological examination. Among six patients, one case belonged to stage Ⅲ, and the others were in stage ⅣA. One case abandoned treatment; one case received chemotherapy; one case underwent supraglottic hemilaryngectomy and adjuvant chemoradiotherapy; one case underwent induction chemotherapy, radiotherapy and consolidation chemotherapy. Two cases received induction chemotherapy, concurrent chemoradiation and consolidation chemotherapy. The drug regimens included bleomycin, fluorouracil,cisplatin, etoposide and taxel for 3 -6 cycles. The radiotherapy technique included conventional radiotherapy, CT-Sim and three dimensional conformal radiation therapy with 60Co or 4 MV X-ray for 60 -66 Gy during 6 - 7 weeks. Results The time of follow-up was 3 - 24 months and the median was 13 months. Two patients applied with concurrent chemoradiation were alive without tumor. The patient abandoning therapy died of respiratory failure, and the others died of lung or liver metastasis after 8 -12 months. Conclusions PSCCL is a disseminated disease, so the pretreatment evaluation is nessesary.Concurrent chemoradiation is an ideal treatment model for this disease.

Non-human primates (NHPs) are confirmed as reservoirs of Cryptosporidium spp., Giardia intestinalis, and Enterocytozoon bieneusi. In this study, 197 fresh fecal samples from 8 NHP species in Qinling Mountains, northwestern China, were collected and examined using multilocus sequence typing (MLST) method. The results showed that 35 (17.8%) samples were positive for tested parasites, including Cryptosporidium spp. (3.0%), G. intestinalis (2.0%), and E. bieneusi (12.7%). Cryptosporidium spp. were detected in 6 fecal samples of Macaca mulatta, and were identified as C. parvum (n=1) and C. andersoni (n=5). Subtyping analysis showed Cryptosporidium spp. belonged to the C. andersoni MLST subtype (A4, A4, A4, and A1) and C. parvum 60 kDa glycoprotein (gp60) subtype IId A15G2R1. G. intestinalis assemblage E was detected in 3 M. mulatta and 1 Saimiri sciureus. Intra-variations were observed at the triose phosphate isomerase (tpi), beta giardin (bg), and glutamate dehydrogenase (gdh) loci, with 3, 1, and 2 new subtypes found in respective locus. E. bieneusi was found in Cercopithecus neglectus (25.0%), Papio hamadrayas (16.7%), M. mulatta (16.3%), S. sciureus (10%), and Rhinopithecus roxellana (9.5%), with 5 ribosomal internal transcribed spacer (ITS) genotypes: 2 known genotypes (D and BEB6) and 3 novel genotypes (MH, XH, and BSH). These findings indicated the presence of zoonotic potential of Cryptosporidium spp. and E. bieneusi in NHPs in Qinling Mountains. This is the first report of C. andersoni in NHPs. The present study provided basic information for control of cryptosporidiosis, giardiasis, and microsporidiosis in human and animals in this area.
Animals ; China ; Cryptosporidiosis/*parasitology ; Cryptosporidium/classification/genetics/*isolation & purification ; Enterocytozoon/classification/genetics/*isolation & purification ; Feces/parasitology ; Female ; Genotype ; Giardia lamblia/classification/genetics/*isolation & purification ; Giardiasis/parasitology/*veterinary ; Male ; Microsporidiosis/parasitology/*veterinary ; Molecular Sequence Data ; Phylogeny ; Primate Diseases/*parasitology ; Primates/classification/parasitology

Base on the improvement of compound FF16, compatibility of Rhodiola crenulata, Cordyceps militaris, and Rheum palmatum, on both insulin resistance and obesity, its effects on type 2 diabetes (T2DM ) was investigated here. The results showed that the levels of fasting and no-fasting blood glucose were controlled in the spontaneous type 2 diabetes KKAy mice; the impaired glucose tolerance (IGT)was improved by decreasing significantly the values of the glucose peaks and the area under the blood glucose-time curve (AUC ) after glucose-loading in glucose tolerance test (OGTT) in both high-fat-diet-induced pre-diabetes IRF mice and KKAy mice, respectively. The pancreatic histopathological analysis showed that the increased islet amount, the enlarged islet area, and the lipid accumulation in the pancreas were reversed by FF16 treatment in both IRF mice and KKAy mice. In the palmitate-induced RINm5f cell model, FF16 could effectively reduce the apoptosis and enhance the glucose-stimulated insulin secretion, respectively. In conclusion, FF16 could improve the T2DM by protecting the pancreatic beta-cells.
Animals ; Blood Glucose ; metabolism ; Cordyceps ; chemistry ; Diabetes Mellitus, Type 2 ; drug therapy ; metabolism ; Drug Compounding ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Insulin Resistance ; Lipid Metabolism ; Metabolic Syndrome ; drug therapy ; metabolism ; Mice ; Mice, Inbred C57BL ; Rheum ; chemistry ; Rhodiola ; chemistry

Aim To investigate the anti-inflammatory effect of L-Shikonin ( SK ) on lipopolysaccharide ( LPS)-induced RAW 264. 7 macrophages in vitro and its protective effect on LPS/D-GalN-induced acute liver injury. Methods The mouse model of acute liver in¬jury was established in vivo experiments by LPS/D- GalN. The survival rate of the mice and the changes of liver and spleen indices in each group were examined. The levels of AST, ALT and AKP in serum and NO, superoxide dismutase ( SOD ) and malondialdehyde (MDA) in liver tissue homogenate were measured, and the histopathological sections of the liver of each group were observed by H&E staining. M I T colorimet- ric assay was used for cell viability in vitro experi¬ments, Griess method for the detection of NO content, RT-PCR assay and Western blot assay for examining the effect of levulinic acid on the expression levels of mRNA and related pathway proteins of pro-inflammato¬ry factors in LPS-induced RAW264. 7 cells. Results The results of in vivo experiments showed that L-SK significantly improved the liver and spleen indices, de¬creased AST, ALT and AKP levels in serum, de¬creased NO and MDA in liver homogenate, and in¬creased SOD activity in mice with acute liver injury. The results of in vitro experiments showed that L-SK significantly inhibited the mRNA expression of INOS, COX2, I FN-(3 and pro-inflammatory factors 1L-6, TNF-a and IL-10 in LPS-induced RAW264. 7 cells, and significantly inhibited the protein expression of IN¬OS, COX2 and the NF-kB signaling pathway. Conclu¬sions L-SK has good anti-inflammatory effects in LPS-induced inflammation in RAW 264. 7 cells in vitro. Il inhibits the protein expression of phosphorylated P65 and IKKaαβ in the NF-kB signaling pathway, thereby suppressing the anti-inflammatory effects in vitro and L- Shikonin has protective effects against acute liver injury in mice.

OBJECTIVE: To explore the potential mechanism of resistance to axitinib in clear cell renal cell carcinoma (ccRCC), with a view to expanding the understanding of axitinib resistance, facilitating the design of more specific treatment options, and improving the treatment effectiveness and survival prognosis of patients. METHODS: By exploring the half maximum inhibitory concentration (IC₅₀) of axitinib on ccRCC cell lines 786-O and Caki-1, cell lines resistant to axitinib were constructed by repeatedly stimulated with axitinib at this concentration for 30 cycles in vitro. Cell lines that were not treated by axitinib were sensitive cell lines. The phenotypic differences of cell proliferation and apoptosis levels between drug resistant and sensitive lines were tested. Genes that might be involved in the drug resistance process were screened from the differentially expressed genes that were co-upregulated in the two drug resistant lines by transcriptome sequencing. The expression level of the target gene in the drug resistant lines was verified by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB). The expression differences of the target gene in ccRCC tumor tissues and adjacent tissues were analyzed in the Gene Expression Profiling Interactive Analysis (GEPIA) public database, and the impact of the target gene on the prognosis of ccRCC patients was analyzed in the Kaplan-Meier Plotter (K-M Plotter) database. After knocking down the target gene in the drug resistant lines using RNA interference by lentivirus vector, the phenotypic differences of the cell lines were tested again. WB was used to detect the levels of apoptosis-related proteins in the different treated cell lines to find molecular pathways that might lead to drug resistance. RESULTS: Cell lines 786-O-R and Caki-1-R resistant to axitinib were successfully constructed in vitro, and their IC₅₀ were significantly higher than those of the sensitive cell lines (10.99 μmol/L, P < 0.01; 11.96 μmol/L, P < 0.01, respectively). Cell counting kit-8 (CCK-8) assay, colony formation, and 5-ethynyl-2 '-deoxyuridine (EdU) assay showed that compared with the sensitive lines, the proliferative ability of the resistant lines decreased, but apoptosis staining showed a significant decrease in the level of cell apoptosis of the resistant lines (P < 0.01). Although resistant to axitinib, the resistant lines had no obvious new replicated cells in the environment of 20 μmol/L axitinib. Nuclear protein 1 (NUPR1) gene was screened by transcriptome sequencing, and its RNA (P < 0.0001) and protein expression levels significantly increased in the resistant lines. Database analysis showed that NUPR1 was significantly overexpressed in ccRCC tumor tissue (P < 0.05); the ccRCC patients with higher expression ofNUPR1had a worse survival prognosis (P < 0.001). Apoptosis staining results showed that knockdown ofNUPR1inhibited the anti-apoptotic ability of the resistant lines to axitinib (786-O, P < 0.01; Caki-1, P < 0.05). WB results showed that knocking downNUPR1decreased the protein level of B-cell lymphoma-2 (BCL2), increased the protein level of BCL2-associated X protein (BAX), decreased the protein level of pro-caspase3, and increased the level of cleaved-caspase3 in the resistant lines after being treated with axitinib. CONCLUSION ccRCC cell lines reduce apoptosis through theNUPR1 -BAX/ BCL2 -caspase3 pathway, which is involved in the process of resistance to axitinib.
Humans ; Carcinoma, Renal Cell/metabolism* ; Axitinib/pharmacology* ; Kidney Neoplasms/metabolism* ; bcl-2-Associated X Protein ; Nuclear Proteins ; Cell Line, Tumor ; Apoptosis ; Cell Proliferation