Objective To investigate the expression level of endogenetic nitric oxide(NO) in the reproduction process of human anterior cruciate ligament cells. Methods Anterior cruciate ligament cells were isolated and subcultured from the human anterior cruciate ligament. LPS was used to induce the anterior cruciate ligament cell to express the inducible nitric oxide synthase(iNOS ), and N-monomethyl -L-Arginine (L-NMMA) was used as the interdiction of nitric oxide. They were alone or together added in the culture medium of anterior cruciate ligament cells in different groups. The level of NO was indirectly measured in the medium of HACL. Results LPS promoted significantly anterior cruciate ligament cells to produce endogenetic nitric oxide, compared with the control group(P

BACKGROUND: Type Ⅱ collagen has been used as the carrier for chondrocyte transplantation in animal models, but whether type Ⅱ collagen may cause arthritis or mediate cytotoxicity remains unknown.OBJECTIVE: To detect the cellular immune functions of the New Zealand rabbits immunized by porcine type Ⅱ collagen.DESIGN: An exploratory comparative study based on the observations.SETTING: An institute of trauma surgery of a municipal hospital.MATERIALS: The study was conducted in the Institute of Trauma Surgery,Guangzhou Red Cross Hospital from August 1999 to February 2000. Six New Zealand rabbits, whose body mass ranged from 2.0 kg to 3.0 kg, were chosen of either gender.METHODS: The rabbits were immunized by porcine type Ⅱ collagen for 60days, during which the plasma was regularly taken for detection of type Ⅱ collagen antibody. On the 60th day, the peripheral blood as well as the spleens and lymph nodes were taken to separate the lymphocytes, which were subjected to secondary stimulation with type Ⅱ collagen in vitro to observe the reactive cell proliferation. The lymphocytes were randomly divided into two groups, and the first group was treated with phytohemagglutinin(PHA) of different concentrations to serve as the positive control, in which non-specific immunity was examined; The second group was treated with type Ⅱ collagen of different concentrations for examining specific immunity.peripheral blood lymphocytes of normal and immunized rabbits.RESULTS: On the 21st day, the titer of the antibody presented the first peak, and 40 days after the re-injection of the antigen the second peak appeared, which maintained for 20 days and then gradually descended. The lymphocytes of the normal rabbits proliferated in response to PHA stimulation but not to the first stimulation with the type Ⅱ collagen. The lymphocytes of the immunized rabbits exhibited significant proliferation upon stimulations with both PHA and type Ⅱ collagen. At the concentration of 25 mg/L, type Ⅱ collagen stimulation was sufficient to induce lymphocyte proliferation, the peak of which occurred when the collagen concentration reached 50 mg/L.CONCLUSION: Xenogenic type Ⅱ collagen at an adequate concentration may induce the increase of the type Ⅱ collagen antibody in immunized rabbits and proliferation of lymphocytes of the spleens and peripheral blood to cause cellular immune reaction and even immunological arthritis in relation to the transplantation.

BACKGROUND: Intra-articular injection of hormone has been applied in the treatment of arthritis because it can alleviate arthralgia rapidly, which is accompanied commonly by progressive cartilage impairments. It is not clear if supplement of growth factor like insulin effect can play a protective role in articular chondrocytes.OBJECTIVE: To observe the effects of insulin or hydrocortisone alone and the combination on the proliferation of chondrocytes.DESIGN: Grouping comparative study, the effect of one medicine was analyzed by using one-factor analysis of variance, while the combined effect was analyzed with multi-factor analysis of variance.SETTING: Guangdong Institute of Trauma Sugery.MATERIALS: Articular cartilage from the knees of New Zealand white rabbits of 4 - 6 weeks old.METHODS: This study was carried out at Guangzhou Traumatic Research Institute from Feberary 2000 to May 2001. Chondrocytes were isolated from the knee joints of New Zealand white rabbits, digested with hyaluronidase,pancreatin and type Ⅱ collagenase and exposed to insulin, hydrocortisone or the combination of insulin and hydrocortisone of different dosage. They were divided into four groups:Control group ( without adding insulin and hydrocortisone), insulin group (0. 035,0. 35,3.5,35 mg/L subgroups), hydrocortisone group(1,5,10,50,100 mg/L subgroups) and insulin(0. 35 mg/L) combined with hydrocortisone(50 mg/L) group. Their influence on chondrocytes proliferation was observed with methyl thiazolyl tetrazolium(MTT) method.sulin.at the concentration of 0. 035 mg/L( P ＜ 0.01 ), reaching the maximum at could inhibit the proliferation of chondrocytes ( P ＜ 0.05 ), which became significant with increasing concentration and no viable chondrocytes could be exposed to 0 . 35 mg/L insulin combined with 50 mg/L hydrocortisone, the promoting effect of insulin was inhibited due to negative cooperation.CONCLUSION: Insulin at low concentration could enhance the proliferation of chondrocytes, but hydrocortisone displayed inhibiting effect on the growth of chondrocytes. The function of insulin was antagonized when combined with hydrocortisone.

The expensive production of bioethanol is because it has not yet reached the 'THREE-HIGH' (High-titer, high-conversion and high-productivity) technical levels of starchy ethanol production. To cope with it, it is necessary to implement a high-gravity mash bioethanol production (HMBP), in which sugar hydrolysates are thick and fermentation-inhibitive compounds are negligible. In this work, HMBP from an atmospheric glycerol autocatalytic organosolv pretreated wheat straw was carried out with different fermentation strategies. Under an optimized condition (15% substrate concentration, 10 g/L (NH4)2SO4, 30 FPU/g dry matter, 10% (V/V) inoculum ratio), HMBP was at 31.2 g/L with a shaking simultaneous saccharification and fermentation (SSF) at 37 degrees C for 72 h, and achieved with a conversion of 73% and a productivity of 0.43 g/(L x h). Further by a semi-SFF with pre-hydrolysis time of 24 h, HMBP reached 33.7 g/L, the conversion and productivity of which was 79% and 0.47 g/(L x h), respectively. During the SSF and semi-SSF, more than 90% of the cellulose in both substrates were hydrolyzed into fermentable sugars. Finally, a fed-batch semi-SFF was developed with an initial substrate concentration of 15%, in which dried substrate (= the weight of the initial substrate) was divided into three portions and added into the conical flask once each 8 h during the first 24 h. HMBP achieved at 51.2 g/L for 72 h with a high productivity of 0.71 g/(L x h) while a low cellulose conversion of 62%. Interestingly, the fermentation inhibitive compound was mainly acetic acid, less than 3.0 g/L, and there were no other inhibitors detected, commonly furfural and hydroxymethyl furfural existing in the slurry. The data indicate that the lignocellulosic substrate subjected to the atmospheric glycerol autocatalytic organosolv pretreatment is very applicable for HMBP. The fed-batch semi-SFF is effective and desirable to realize an HMBP.
Biofuels ; Carbohydrates ; chemistry ; Cellulose ; chemistry ; Ethanol ; metabolism ; Fermentation ; Furaldehyde ; chemistry ; Glycerol ; chemistry ; Hydrolysis ; Triticum

Objective To investigate the relationship between the degree of liver fibrosis in non-alcoholic fatty liver disease(NAFLD)and the left ventricular structure and function of the heart.Methods A total of 411 patients with NAFLD admitted to the Sec-ond Affiliated Hospital of Xinjiang Medical University from June 2022 to April 2023 were collected.NAFLD patients were stratified using the FIB-4 non-invasive liver fibrosis scoring system and divided into the progressive fibrosis group(n=138)and non-progressive fi-brosis group(n=273).A total of 151 patients with normal abdominal ultrasound examination during the same period were selected as the control group,the relationship between the severity of liver fibrosis in NAFLD patients and the left ventricular structure and function of the heart were analyzed.Results Liver fibrosis was a risk factor for cardiac diastolic function limitation in NAFLD patients(P＜0.05),and the more severe the degree of liver fibrosis,the more severe the cardiac diastolic function limitation.Conclusion The degree of liver fi-brosis in NAFLD is significantly correlated with limited left ventricular diastolic function;it may be related to changes in the left ventricu-lar structure of the heart.

To detect the cellular immunity state of New Zealand white rabbit immunized by pig type II collagen. The New Zealand white rabbit was immunized by type II collagen for sixty days. The plasma was collected at a regular interval and the anti-type II collagen antibodies were examined. At the sixtieth day, the peripheral circular lymphocytes and the lymphocytes separated from spleen cells of rabbit and lymph nodes were collected and were stimulated by type II collagen in vivo again. The regulation of reactive cellular proliferation caused by the stimulation was detected. The experiment samples were divided into two groups. The first group was the positive control group by adding different concentrations of PHA and the non-specific immunity was assayed. The different concentrations of type II collagen were added to the second group and the specific immunity was assayed. The lymphocytes of normal rabbits showed proliferation by PHA stimulation but no proliferation by the first stimulation of type II collagen. Obvious proliferation due to the stimulation of both PHA and type II collagen in the immunized rabbit were observed. It shows that certain concentration of heterogeneous collagen may cause an increase of anti-type II collagen antibody in immunized rabbit and may cause a proliferation of lymphocytes in rabbit spleen and peripheral blood. The heterogeneous type II collagen causes cellular immunity in vivo.
Animals ; Cell Division ; drug effects ; Collagen Type II ; administration & dosage ; immunology ; Cytotoxicity, Immunologic ; Female ; Histocompatibility Antigens ; Lymphocytes ; cytology ; immunology ; Male ; Rabbits ; Spleen ; cytology ; Swine ; Transplantation, Heterologous

Pancreatic cancer has a very poor prognosis.Early diagnosis and treatment are especially critical for improving its prognosis.Nanotechnology has been widely used in the diagnosis and treatment of pancreatic cancer.Relying on the unique physicochemical properties of nanoparticles and their rich surface modifications,effective enrichment of tumor sites can be achieved.Magnetic iron oxide nanoparticles(MIONPs)is one of the commonly used nanomaterials in the diagnosis and treatment of pancreatic cancer,and has good biocompatibility.Through special surface modification,it can be used in targeted diagnosis and treatment of pancreatic cancer.MIONPs can be used as a contrast agent for MRI,and by modifying the surface,they also can be used in targeted imaging of pancreatic cancer.And they can also be modified as a drug delivery system to achieve targeted delivery of drugs and improve therapeutic effects.However,the application of MIONPs in pancreatic cancer diagnosis and treatment still faces some challenges,such as nanotoxicity and cost issues.With the development of technology,MIONPs are expected to play an important role in the personalized diagnosis and treatment of pancreatic cancer.

Objective To investigate the characteristics of amygdala neural circuitry in comorbidity of late-life depression (LLD) and cognitive impairment. Methods Twenty-four LLD,eighteen amnestic mild cognitive impairments (aMCI),thirteen aMCI with depression (dMCI) and thirty cognitive normal (CN) subjects completed resting-state functional magnetic resonance imaging scan. Main effects of depression and MCI and their interactions on the intrinsic amygdala functional connectivity network ( AFCN) connectivity were examined. Behavioral significance of AFCN that voxel-wised amygdala connectivity correlating with de-pression severity and memory scores were also tested after controlling the effects of covariates,including age, gender,education, gray matter atrophy, and group. Results The immediate memory and delayed memory function in the aMCI group (-0. 75 ± 0. 77 and -1. 13 ± 0. 56) and the dMCI group (-1. 07 ± 0. 79 and-1. 00±0. 52) were significantly lower than those of the CN group (0. 46±0. 73 and 0. 60±0. 61),and the difference was statistically significant (P<0. 01). Depression and anxiety in the LLD group (1. 00±0. 53 and 0. 93±0. 98) and the dMCI group (0. 86±0. 80 and 0. 78±0. 82) were significantly higher than those of the CN group (-0. 92±0. 25 and -0. 74±0. 22),and the difference was statistically significant (P<0. 01). Brain network analysis showed that separated neural circuits were implicated in the depression and cognitive im-pairment. Importantly,interactive effects of depression and MCI on the AFCN were also identified,especially in the bilateral somatomotor area,inferior parietal cortex/precuneus,posterior cingulate cortex,right medial prefrontal cortex/dorsolateral prefrontal cortex and hippocampus. Behavioral significance of AFCN also re-vealed the distinctive neural circuits involved in the depression severity and memory deficits,respectively. Conjunction analysis further identified the overlapped neural circuits associated with depression and memory deficits were primarily in the left DLPFC,insula,hippocampus,right inferior prefrontal cortex and dorsomedi-al prefrontal cortex. Conclusions Depression and cognitive impairment synergistically facilitate functional decoupling of AFCN and thus compromise the integrity of amygdala networks. Distinct depression-related or MCI-related neural constructs represent the characteristics of clinical phenotype of depression or MCI alone, while overlapped circuits probably reveal the neural basis of comorbidity of LLD and MCI.

【Objective】 To investigate the effects of resveratrol on gemcitabine chemotherapy in pancreatic cancer and the possible molecular mechanism. 【Methods】 Gemcitabine resistant cell lines were screened by continuous low concentration increasing induction. High-throughput RNA-seq was used to analyze the differential expression enrichment pathway, COMET assay was used to detect DNA damage, Western blotting was used to detect related pathway indicators, and Chou-Talalay was used to calculate drug combination synergistic index. AutoDock predicts docking targets for small molecules and proteins. 【Results】 DNA damage repair related pathways were activated in drug-resistant cell lines compared with their parents. Resveratrol enhanced the DNA damage effects induced by gemcitabine (P<0.01). Resveratrol inhibited the expression of PARP1, a key molecule of DNA damage repair, and played a synergic effect with gemcitabine (CI<1). Resveratrol has docking targets with the CAT domain of PARP1. 【Conclusion】 Resveratrol can inhibit PARP1, a key molecule of chemotherapy resistance, and has a synergistic effect with gemcitabine in pancreatic cancer chemotherapy.

As one of the hallmarks of cancer, metabolic reprogramming leads to cancer progression, and targeting glycolytic enzymes could be useful strategies for cancer therapy. By screening a small molecule library consisting of 1320 FDA-approved drugs, we found that penfluridol, an antipsychotic drug used to treat schizophrenia, could inhibit glycolysis and induce apoptosis in esophageal squamous cell carcinoma (ESCC). Gene profiling and Ingenuity Pathway Analysis suggested the important role of AMPK in action mechanism of penfluridol. By using drug affinity responsive target stability (DARTS) technology and proteomics, we identified phosphofructokinase, liver type (PFKL), a key enzyme in glycolysis, as a direct target of penfluridol. Penfluridol could not exhibit its anticancer property in PFKL-deficient cancer cells, illustrating that PFKL is essential for the bioactivity of penfluridol. High PFKL expression is correlated with advanced stages and poor survival of ESCC patients, and silencing of PFKL significantly suppressed tumor growth. Mechanistically, direct binding of penfluridol and PFKL inhibits glucose consumption, lactate and ATP production, leads to nuclear translocation of FOXO3a and subsequent transcriptional activation of BIM in an AMPK-dependent manner. Taken together, PFKL is a potential prognostic biomarker and therapeutic target in ESCC, and penfluridol may be a new therapeutic option for management of this lethal disease.