Bioprocess equipment is of great importance in application of modern industrial biotechnology.With the rapid development of industrial biotechnology,demands for talents capable of understanding the theory,design and manipulation of modern bioprocess equipment increased.The experiences in aspects such as the building of teachers' contingent,construction of teaching materials,innovation of teaching method from the top-quality course construction of Bioprocess Equipment was discussed in this paper.

The drinking behavior and the c-fos expression in rat brain induced by electrical stimulation of the subfornical organ (SFO) were examined. SFO stimulation induced stable and significant drinking behavior and Fos protein expression in 8 areas of the forebrain (organum vasculosum of the lamina terminalis, median preoptic nucleus, paraventricular nucleus, supraoptic nucleus, lateral hypothalamic area, perifornical dorsal area, substantia innominata and thalamic reuniens nucleus), and in 3 areas of the hindbrain (area postrema, solitary tract nucleus and lateral parabrachial nucleus). In certain neurons of paraventricular and supraoptic nuclei, co-expression of Fos protein and vasopressin was induced by SFO stimulation. Intracerebroventricular injection of atropine partly blocked the SFO stimulation-induced drinking behavior and the Fos protein expression in the brain, suggesting that an M-cholinergic mechanism may be involved.

NMDA-induced excitotoxicity cause severe neuronal damage including apoptosis and necrosis. The present study was aimed to evaluate the proportion of NMDA-induced apoptosis of rat cortical neurons and discover signal transduction mechanism. Caspase inhibitor and lactate dehydrogenase (LDH) assay were used to study the NMDA-induced apoptosis. To explore the involved signal pathways, the primary culture of rat cortical neurons were pretreated by the inhibitors of three MAPK pathways, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. With 2 h of NMDA treatment, cellular apoptosis was measured by caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) and Annexin V staining. The results showed that: (1) Caspase-dependent apoptosis accounted for 22.49% in NMDA-induced neuronal death; (2) Pretreatment with p38 MAPK inhibitor SB203580 (10 μmol/L) significantly decreased NMDA-mediated caspase-3 activity by 30.43% (P < 0.05). However, ERK inhibitor PD98059 (20 μmol/L) or JNK inhibitor SP600125 (20 μmol/L) did not influence caspase-3 activity; (3) Pretreatment with SB203580 significantly reduced the number of NMDA-induced TUNEL-positive cells by 33.10% (P < 0.05). PD98059 (20 μmol/L) or SP600125 (20 μmol/L) did not show obvious effect; (4) Pretreatment with SB203580 (10 μmol/L) significantly reduced the number of NMDA-induced early apoptotic neurons by 55.56% (P < 0.05). Also, SP600125 (20 μmol/L) significantly decreased the amount of late apoptotic/dead cells by 67.59% (P < 0.05). There was no effect of PD98059 (20 μmol/L). These results indicate that: (1) NMDA induces neuronal apoptosis besides necrosis; (2) p38 MAPK, but not JNK and ERK, is involved in NMDA-induced neuronal apoptosis, and inhibition of the apoptotic signaling pathway contributes to neuroprotection; (3) JNK activation might contribute to NMDA-induced neuronal necrosis rather than apoptosis.
Animals ; Anthracenes ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; N-Methylaspartate ; pharmacology ; Neurons ; cytology ; Primary Cell Culture ; Pyridines ; pharmacology ; Rats ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

OBJECTIVETo study mechanism of the apoptosis of rat pancreas islet β cell strain (INS-1 cells) induced by sodium arsenite.

METHODSINS-1 cells were exposed to sodium arsenite at the different concentrations. MTT assay was used to detect the viability of INS-1 cells. The potentials on mitochondrial membrane and lysosome membrane of INS-1 cells were determined with the fluorescence spectrophotometer. The apoptotic levels of INS-1 cells exposed to sodium arsenite were observed by a fluorescence microscope and flow cytometry.

RESULTSAfter exposure to sodium arsenite, the viability of INS-1 cells significantly decreased with the doses of sodium arsenite. At 24 h after exposure, the OD values of the mitochondrial membrane potentials declined observably with the doses of sodium arsenite (P < 0.01). At 48 h after exposure, the OD values of the lysosome membrane potentials significantly increased with the doses of sodium arsenite (P < 0.01). At 72 h after exposure, the apoptotic cells were observed under a fluorescence microscope and enhanced with the doses of sodium arsenite. The apoptosis cells with light blue, karyopyknosis, karyorrhexis, apoptotic body and chromatin concentration appeared. The results detected with flow cytometry indicated that after exposure, the apoptotic INS-1E cells significantly increased with the doses of sodium arsenite.

CONCLUSIONSThe sodium arsenite can induce the apoptosis of INS-1 cells through the mitochondria-lysosome pathway.

Animals ; Apoptosis ; drug effects ; Arsenites ; toxicity ; Cells, Cultured ; Insulin-Secreting Cells ; drug effects ; Lysosomes ; metabolism ; Membrane Potentials ; drug effects ; Mitochondria ; metabolism ; Rats ; Sodium Compounds ; toxicity

OBJECTIVETo construct mouse enhanced green fluorecence protein (EGFP) -peroxisome proliferator-activated receptor (PPAR)gamma2, and to detect EGFP-PPARgamma2 expression in infected mouse bone marrow mesenchymal stem cells (BMSC).

METHODSCut the fragment of PPARgamma2 from the expression plasmid pcDNA flag PPARgamma2, then cloned the gene fragment into pEGFP-C1 and pEGFP-N1 vector. Subsequently, subclone the fragment EGFP-PPARgamma2 from pEGFP-C1-PPARgamma2 into the shuttle plasmid DC315. HEK293 cells were co-transfected with the constructed recombinant shuttle plasmid DC315-EGFP-PPARgamma2 and large adenovirus helper plasmid pBHGlox deltaE1, 3Cre in mediation of liposome. The obtained replication-defective recombinant adenovirus Ad-EGFP-PPARgamma2 was confirmed. Then it was propagated in HEK293 cells. After the BMSC were transfected for 72 h, adipogenic differentiation was demonstrated.

RESULTSHEK293 cells were transfected with the pEGFP-C1-PPARgamma2 or pEGFP-N1-PPARgamma2 in mediation of liposome. The former green fluorescence protein was better than the latter by fluorescence microscope. The recombinant plasmids were digested and identified. Western blot analysis showed the expression of EGFP-PPARgamma2 in vitro. EGFP-PPARgamma2 protein was detectable in the nucleus of BMSC.

CONCLUSIONThe recombinant adenovirus encoding EGFP-PPARgamma2 fusion protein was successfully constructed, which provided a basis for application of EGFP-PPARgamma2 gene to adenovirus-mediated gene therapy.

Adenoviridae ; Animals ; Bone Marrow Cells ; metabolism ; Genetic Vectors ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Mesenchymal Stromal Cells ; metabolism ; Mice ; PPAR gamma ; metabolism ; Recombinant Proteins ; metabolism ; Transfection

measured by OCT 1 and 3mo after surgery. Chi-square test was used to compare preoperative and postoperative BCVA. Pearsion’s correlation was used to evaluate relationship between postoperative BCVA and central foveal thickness.RESULTS:The ratio of BCVA<0. 05 was 30%,14%, 11%and 7% respectively for per-operation, 1wk, 1 and 3mo post - operation. After surgery, the central foveal thickness was significantly increased in group with BCVA<0. 3 comparing to group with BCVA≥0. 3. Three month post-operation, central foveal thickness was significantly decreased in both groups comparing to that in 1mo post-operation (P<0. 01). There has significant correlation between 3mo postoperative BCVA and central foveal thickness (r=-0.716, P<0.05).CONCLUSlON: ln this study, BCVA is improved after 3mo follow up. There has significant correlation between postoperative BCVA and central foveal thickness.

Objective:To investigate the indications and effects of arthroscopic all-inside reconstruction in the treatment of isolated posterior cruciate ligament (PCL) injury.Methods:A retrospective analysis was performed on 47 patients with isolated PCL injury, who underwent arthroscopic all-inside reconstruction in the Third Medical Center of the PLA General Hospital from January 2016 to January 2020. There were 39 males and 8 females, aged 27.14±7.70 years old (range 16-40 years old). The preoperative kneeling-position stress X-ray showed that the degree of tibial posterior displacement was 8-10 mm, which was a complete and isolated Grade II PCL injury. The tibial and femoral tunnels were created through posterior-medial, anteromedial, and anterolateral portals, while the lateral portal to the medial femoral condyle was enlarged to position the tibial tunnel and protect the anterior cruciate ligament. The autologous graft tendon was pulled through the femoral and tibial tunnels secured with an adjustable loop plate. The efficacy was evaluated by evaluating and comparing preoperative and postoperative Lachman test, posterior drawer test, knee range of motion and relaxation, pain visual analogue scale (VAS) and Lysholm score.Results:43 patients were followed up for 35.21±3.88 months (range 12-40 months). The symptoms of knee instability all improved after surgery. At the follow-up of 1 year after surgery, 41 (95%) and 40 (93%) patients showed normal or I-degree laxity in Lachman test and posterior drawer test, respectively. The active range of motion and passive flexion of the knee joint were increased to 90°-110° and 110°-130°, respectively. The Lysholm score was 86.44±4.08 at the first year of follow-up and 90.12±3.33 at the last follow-up with significant difference compared with pre-operations ( P<0.05). The VAS score was 2.07±0.94 at the first year of follow-up and 1.28±0.83 at the last follow-up with significant difference compared with pre-operations ( P<0.05). The Lysholm score and VAS were 90.12±3.33 and 1.28±0.83, which were significantly improved compared to 1-year-follow-up ( P<0.05). Conclusion:Routine kneeling stress X-rays can evaluate the degree of tibial posterior displacement in isolated PCL injuries. With tibial posterior displacement equal to or greater than 10 mm, surgical reconstruction was required. All-inside reconstruction of isolated PCL injury was a safe and minimally invasive surgery to improve symptoms and restore knee functions.

AIMTo explore the effects of periphery injection of L-SOP on the activation of p38MAPK in spinal cord in formalin pain model in rats.

METHODSFourty-eight male Wistar rats were divided randomly into four groups (n=12): NS group and three different dose of L-SOP groups. For each group, 6 rats used to observe flinching and licking time every as nociception behavior 3 minutes in 1 hour after formalin injected and the other 6 rats used to observe the activation of p38(P-p38) by Western blotting.

RESULTSAll the three different groups of L-SOP could inhibit nociception behavior in the tonic phase,and 250 nmoVl/L and 500 nmol/L groups could suppress not only in the tonic phase but also in the acute phase. 250 nmol/L and 500 nmol/L groups could reduce activated or phosphorylated p38MAPK in spinal cord.

CONCLUSIONPeriphery injection of L-SOP can reduce nociceptive behavior and phosphorylated p38MAPK in the spinal cord in formalin-induced hyperalgia, it is suggested that there is functional expression of mGluRs III in the periphery and is involved in the processing of peripheral noxious informations.

Animals ; Formaldehyde ; Male ; Nociception ; physiology ; Pain ; chemically induced ; metabolism ; physiopathology ; Phosphoserine ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Metabotropic Glutamate ; physiology ; Spinal Cord ; metabolism ; physiopathology ; p38 Mitogen-Activated Protein Kinases ; metabolism

Silent information regulator 1 (SIRT1), an NAD(+)-dependent deacetylase, is involved in the regulation of gene transcription, energy metabolism and cell aging. Recent studies have showed that SIRT1 possesses neuroprotective effects, however, it is not very clear how SIRT1 exerts the neuroprotection in Alzheimer's disease (AD). In this review, we summarized the neuroprotective role of SIRT1 in AD and its possible molecular mechanisms, proposing a novel strategy for preventing and treating neurodegeneration.
Alzheimer Disease ; genetics ; physiopathology ; Animals ; Energy Metabolism ; physiology ; Humans ; Neuroprotective Agents ; Sirtuin 1 ; physiology ; Transcription, Genetic ; physiology

OBJECTIVETo evaluate the accuracy of sentinel lymph node mapping(SLM) in patients with rectal cancer by single-photon emission computed tomography (SPECT-CT) lymphoscintigraphy and carbon nanoparticles suspension injection.

METHODSTwelve patients with clinical T(1-2)N(0)M(0) rectal cancer were selected and locally injected with technetium-(99m)sulfur-colloid and carbon nanoparticles suspension by endoscope one day before surgery, followed by SPECT-CT scanning 1, 3 and 5 hours later. Radioactive isotope(RI) uptake of each sentinel node(SN) basin with location preoperatively determined by SPECT-CT was postoperatively calculated using gamma probe. Nodes with the highest RI uptake, the number of which was also pre-determined by SPECT-CT, was defined as SNs. Immunohistochemical cytokeratin staining was performed for all the SNs and non-SNs.

RESULTSThe rate of sentinel node detection was 91.7%(11/12) with at least one SN(1-3) per patient. Ten cases showed metastasis-negative in SNs as well as all the resected regional nodes by immunohistochemical cytokeratin staining. Only one patient had positive nodes in both SN and non-SNs. The accuracy of SLM was 100%.

CONCLUSIONSPECT-CT lymphoscintigraphy and carbon nanoparticles suspension injection can effectively detect the anatomic location and number of sentinel nodes, and improve the accuracy of SLM for rectal cancer.

Adult ; Aged ; Carbon ; Female ; Humans ; Male ; Middle Aged ; Nanostructures ; Rectal Neoplasms ; diagnosis ; diagnostic imaging ; pathology ; Sentinel Lymph Node Biopsy ; methods ; Tomography, Emission-Computed, Single-Photon ; methods ; Tomography, X-Ray Computed ; methods