Metformin improves sheep sperm cryopreservation via vitalizing the AMPK pathway.

Adenosine 5'-monophosphate (AMP)-activated protein kinase (AMPK) is a key regulator of sperm function and physiological metabolism. Metformin, an inexpensive and effective antioxidant, is known to play an important role in the activation of AMPK. Therefore metformin has potential to improve sperm cryopreservation. The aim of this study was to investigate the effect of metformin during semen cryopreservation of sheep and to find the most effective concentration in freezing extender. Semen were cryopreserved with extender containing different concentrations of metformin (0, 0.25, 0.5, 1.0, 2.0 and 4.0 mmol/L). Sperm motility, acrosome integrity and plasma membrane integrity were measured after semen freezing and thawing. All results showed that sperm quality was significantly increased in the 1.0 mmol/L metformin-treated group compared with the control group (P < 0.05). In addition, the study showed that metformin effectively reduced the content of malondialdehyde (MDA) and reactive oxygen species (ROS), and increased the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), catalase (CAT) and total antioxidant capacity (T-AOC) of freeze-thawed sperm (P < 0.05). The optimal concentration of metformin was 1.0 mmol/L. Moreover, the results showed that AMPK was localized in the acrosome region, junction and midsection of sperm, and p-AMPK was distributed in the post-acrosomal region, junction and midsection. Western blot analysis indicated that 1.0 mmol/L metformin stimulated the phosphorylation of AMPK in sperm. Further results showed that 1.0 mmol/L metformin significantly increased the mitochondrial membrane potential (ΔΨm), ATP content, glucose uptake and lactate efflux of post-thawed sperm through the AMPK pathway, improved sperm quality, and increased the cleavage rate of in vitro fertilization (P < 0.05).

Dieckol, a natural polyphenolic drug, inhibits the proliferation and migration of colon cancer cells by inhibiting PI3K, AKT, and mTOR phosphorylation.

This study investigated that dieckol (DKL), a natural drug, inhibits colon cancer cell proliferation and migration by inhibiting phosphoinositide-3-kinase (PI3K), protein kinase B (AKT), and mammalian target of rapamycin (mTOR) phosphorylation in HCT-116 cells. The cells were treated with DKL in various concentrations (32 and 50 µM) for 24 h and then analyzed for various experiments. MTT (tetrazolium bromide) and crystal violet assay investigated DKL-mediated cytotoxicity. Dichlorodihydrofluorescein diacetate staining was used to assess the reactive oxygen species (ROS) measurement, and apoptotic changes were studied by dual acridine orange and ethidium bromide staining. Protein expression of cell survival, cell cycle, proliferation, and apoptosis protein was evaluated by western blot analysis. Results indicated that DKL produces significant cytotoxicity in HCT-116, and the half-maximal inhibitory concentration was found to be 32 µM for 24-h incubation. Moreover, effective production of ROS and enhanced apoptotic signs were observed upon DKL treatment in HCT-116. DKL induces the expression of phosphorylated PI3K, AKT, and mToR-associated enhanced expression of cyclin-D1, proliferating cell nuclear antigen, cyclin-dependent kinase (CDK)-4, CDK-6, and Bcl-2 in HCT-116. In addition, proapoptotic proteins such as Bax, caspase-9, and caspase-3 were significantly enhanced by DKL treatment in HCT-116. Hence, DKL has been considered a chemotherapeutic drug by impeding the expression of PI3K-, AKT-, and mTOR-mediated inhibition of proliferation and cell cycle-regulating proteins.

Heterogeneity of tumor microenvironment is associated with clinical prognosis of non-clear cell renal cell carcinoma: a single-cell genomics study.

Non-clear renal cell carcinomas (nccRCCs) are less frequent in kidney cancer with histopathological heterogeneity. A better understanding of the tumor biology of nccRCC can provide more effective treatment paradigms for different subtypes. To reveal the heterogeneity of tumor microenvironment (TME) in nccRCC, we performed 10x sing-cell genomics on tumor and normal tissues from patients with papillary renal cell carcinoma (pRCC), chromophobe RCC (chrRCC), collecting duct carcinoma (CDRCC) and sarcomatoid RCC (sarRCC). 15 tissue samples were finally included. 34561 cells were identified as 16 major cell clusters with 34 cell subtypes. Our study presented the sing-cell landscape for four types of nccRCC, and demonstrated that CD8+ T cells exhaustion, tumor-associated macrophages (TAMs) and sarcomatoid process were the pivotal factors in immunosuppression of nccRCC tissues and were closely correlated with poor prognosis. Abnormal metabolic patterns were present in both cancer cells and tumor-infiltrating stromal cells, such as fibroblasts and endothelial cells. Combined with CIBERSORTx tool, the expression data of bulk RNA-seq from TCGA were labeled with cell types of our sing-cell data. Calculation of the relative abundance of cell types revealed that greater proportion of exhausted CD8+ T cells, TAMs and sarRCC derived cells were correlated with poor prognosis in the cohort of 274 nccRCC patients. To the best of our knowledge, this is the first study that provides a more comprehensive sight about the heterogeneity and tumor biology of nccRCC, which may potentially facilitate the development of more effective therapies for nccRCC.

[Economic burden and related factors on inpatients with HBV-related diseases in Shandong province].

OBJECTIVE: To investigate the economic burden of patients with acute and chronic hepatitis B, cirrhosis and liver cancer caused by hepatitis B virus (HBV). METHODS: Cluster sampling was used on cases consecutively collected during the study period. Questionnaire survey was conducted and information on the expenses during hospitalization was collected from the hospital records and through interviewing those patients. RESULTS: Yearly costs related to patients with acute hepatitis B, severe hepatitis B, chronic hepatitis B, cirrhosis, hepatocellular carcinoma were 66.7, 138.1, 127.4, 151.7 and 377.2 thousand Yuan, respectively. RESULTS: from multiple linear regression model showed that the type of medical insurance scheme, annual days of hospitalization, classifications of HBV-related diseases and personal income were major influencing factors on the cost. CONCLUSION: HBV infection caused considerable burden to families and the society, indicating that HBV infection control programs would bring huge potential benefits. The reform of insurance scheme should be administrated to promote social fairness.

Butanol production from hemicellulosic hydrolysate of corn fiber by a Clostridium beijerinckii mutant with high inhibitor-tolerance.

A Clostridium beijerinckii mutant RT66 with considerable inhibitor-tolerance obtained by continuous culture was used for butanol production from non-detoxified hemicellulosic hydrolysate of corn fiber treated with dilute sulfuric acid (SAHHC). In fed-batch fermentation, 1.8L of diluted SAHHC containing 10 g/L of reducing sugar was provided during the acidogenic phase and 0.2L of concentrated SAHHC containing 300 g/L of reducing sugar was provided during the solventogenic phase. The mutant produced a total amount of solvents of 12.9 g/L, which consisted of 3.1 g/L of acetone, 9.3 g/L of butanol and 0.5 g/L of ethanol. A solvent yield of 0.35 g/g sugar and a productivity of 0.18 g/L h in 72 h were achieved. The remarkable inhibitor-tolerance of C. beijerinckii RT66 demonstrates that this may be an excellent strain for butanol production from ligocellulosic materials.

Acetone-butanol-ethanol (ABE) fermentation using Clostridium acetobutylicum XY16 and in situ recovery by PDMS/ceramic composite membrane.

PDMS/ceramic composite membrane was directly integrated with acetone-butanol-ethanol (ABE) fermentation using Clostridium acetobutylicum XY16 at 37 °C and in situ removing ABE from fermentation broth. The membrane was integrated with batch fermentation, and approximately 46 % solvent was extracted. The solvent in permeates was 118 g/L, and solvent productivity was 0.303 g/(L/h), which was approximately 33 % higher compared with the batch fermentation without in situ recovery. The fed-batch fermentation with in situ recovery by pervaporation continued for more than 200 h, 61 % solvent was extracted, and the solvent in penetration was 96.2 g/L. The total flux ranged from 0.338 to 0.847 kg/(m(2)/h) and the separation factor of butanol ranged from 5.1 to 27.1 in this process. The membrane was fouled by the active fermentation broth, nevertheless the separation performances were partially recovered by offline membrane cleaning, and the solvent productivity was increased to 0.252 g/(L/h), which was 19 % higher compared with that in situ recovery process without membrane cleaning.

[Effects of management regime on soil respiration from agroecosystems].

In order to examine the effects of management regime, such as nitrogen application and plowing method, on soil respiration from farmland, the static opaque chamber-gas chromatograph method was used to measure soil CO2 fluxes in situ. The field measurement was carried out for 5 growing seasons, which were the 2002-2003 wheat, 2003 maize and soybean, 2003-2004 wheat, 2004 maize and 2004-2005 wheat seasons. Our results showed that soil respiration increased in fertilizer-applied treatments compared with no fertilizer treatment after 3 times of fertilizer application on 9 November 2002, 14 February and 26 March 2003. And the most obvious increase appeared following the third fertilizer application. No significant difference in soil respiration was found among several fertilizer application treatments. The effect of plowing depth on soil respiration was contingent on preceding cropping practice. Over the 2003-2004 wheat-growing seasons (its preceding cropping practice was rice paddy), mean soil respiration rates were not significant different (p > 0.05) between no plowing treatment and shallow plowing treatment. The shallow plowing treatment CT2 led to higher soil CO2 losses compared with no plowing treatment of NT2 in the 2004 maize-growing season, however, the significant higher (p < 0.05) soil respiration rates occurred with no plowing treatment of NT3 in the following 2004-2005 wheat-growing season. Intensive plowing (25 cm depth), compared with no plowing practice (NT4), increased soil respiration significantly during the 2004-2005 wheat-growing season. Regression analysis showed that the exponential function could be employed to fit the relationship between soil respiration and temperature. The exponential relationship yielded the Q10 values which were varied from 1.26 to 3.60, with a mean value of 2.08. To evaluate the effect of temperature on soil respiration, the CO2 emission fluxes were normalized for each treatment and each crop growing season. Plotting the normalized soil respiration against the temperature, the exponential relationship between these two parameters becomes obvious. The temperature coefficient Q10 was then evaluated as 1.66 according to the exponential relationship. Further investigation indicated that soil respiration could be well simulated by an empirical model in which the effects of both soil temperature and moisture on soil respiration were considered. This model described 54% variances of the measured 463 soil respiration rates, with a R2 of 0.54 and a p value less than 0.0001.

[Single-lung transplantation and contralateral lung volume reduction for emphysema].

OBJECTIVE: To report the operation technique, patient selection and curative effect of single-lung transplantation with contralateral lung volume reduction for a patient with end-stage emphysema. METHODS: A 47-year-old male patient who was dependent on mechanical ventilation because of end-stage emphysema received left-lung transplantation on 20 June, 2003. The surgical technique used was similar to that mentioned in the literature. The donor lung was perfused by low-potassium dextran solutions (LPD) with a cold ischemic time of 390 minutes. The patient received contralateral lung volume reduction because of subacute native lung hyperinflation on the 7th day after operation. He was received tracheostomy on 15th day after operation and was weaned from ventilator on the 26th day post-operation. Immunosuppression included cyclosporine, mycophenolate mofetil and corticosteroid. The acute rejection occurred on the 9th and 15th days after operation and was cured successfully. RESULTS: The lung function was improved significantly and the patient was discharged from hospital 71 days after operation. CONCLUSIONS: Single-lung transplantation combined with contralateral lung volume reduction for end-stage emphysema is an effective measure for subacute native lung hyperinflation. Further follow-up is required to assess the long term results of this procedure.

[Single-lung transplantation for end-stage emphysema].

OBJECTIVE: To evaluate operative technique, patient selection and perioperative management of single-lung transplantation for a patients with end-stage emphysema. METHODS: A 56-year-old patient with end-stage emphysema underwent left-lung transplantation on September 28, 2002. The surgical technique used was similar to that mentioned in the literature. The donor lung was perfused by LPD solution with a cold ischemic time of 260 minutes. Cardiopulmonary bypass was not performed. RESULTS: The patient weaned from a ventilator at the 93th hour after operation. Immunosuppressants included cyclosporine, mycophenolate mofetil and corticosteroid. Acute rejection occurred on the ninth day after operation and was cured by bolus methylprednisolone given intravenously. Lung function was improved significantly and the patient was discharged from the hospital on the 47th day after operation. CONCLUSION: Single-lung transplantation for patients with end-stage emphysema is effective for long-term improvement of pulmonary function.

[Video-assisted thoracoscopic surgery for spontaneous pneumothorax].

OBJECTIVE: To explore the operational procedures, indications, and the long-term results of video-assisted thoracoscopic surgery (VATS) in the treatment of spontaneous pneumothorax. METHODS: One hundred seventy three cases of spontaneous pneumothorax treated with VATS in our hospital between 1995 and 2001 were analyzed. Primary spontaneous pneumothorax (PSP) was found in 157 cases and secondary spontaneous pneumothorax (SSP) in 16 cases. In patients with PSP, pneumothorax occurred for the first time in 65 (41.4%) cases, and recurred at least once in 92 (58.6%) cases. Among them three patients underwent simultaneous bilateral VATS for simultaneous bilateral pneumothorax, and 6 underwent two-stage bilateral VATS. In patients with SSP, 6 had recurrent pneumothorax. RESULTS: According to the Vanderschueren's classification, no abnormality was found in 34 (19.7%) cases, pleuro-pulmonary adhesions in 41 (23.7%) cases and bullae in 98 (56.7%) cases. Pleurodesis and stapling under Endo-cutter were the major operational procedures used in these cases. The median follow-up time was 53 months. The long-term recurrence rate was 1.8%. Complications related to surgery occurred in 3.0% of the cases, which included subcutaneous emphysema, localized pleural effusion and prolonged air leakage. CONCLUSION: VATS is a safe and effective treatment for both initial and recurrent spontaneous pneumothorax, and therefore it is becoming the primary treatment modality for this disease.