Brown-Sequard Syndrome ; etiology ; Cervical Vertebrae ; Humans ; Intervertebral Disc Displacement ; complications ; surgery ; Male ; Middle Aged

Adult ; Autopsy ; Female ; Hair ; metabolism ; Humans ; Liver ; pathology ; Thallium ; poisoning

Adenocarcinoma, Clear Cell ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Female ; Humans ; Keratins ; metabolism ; Male ; Mucin-1 ; metabolism ; Palatal Neoplasms ; metabolism ; pathology ; surgery ; Palate, Hard ; surgery ; Tongue Neoplasms ; metabolism ; pathology ; surgery

Background Human retinal pigment epithelial (RPE) cell transplantation treating retinal degenerative diseases is a researching topic,and the source of human RPE cells is a key problem.Many biological carriers can be used for the preparation of RPE cell layer.However,some advantages,such as cytotoxicity,lack of stability and immunologic reaction etc.are still existed.To study an ideal biological carrier is very important.Objective This experimental was to determine the effects of amniotic membrane on the proliferation and differentiation of human RPE cells and the possibility as a scaffold for RPE cell transplantation.Methods ARPE19 cell line cells were cultured and passaged in DMEM/F12 medium with 10％ fetal bovine serum,and 8-12generation of cells were used.The cells were divided into two groups.One group of cells were incubated on the denuded amniotic membrane,and the other group of cells were cultured in the medium (control group).MTT was performed to detect the A492 value of RPE cells for the evaluation of cell proliferation ability 24,48,72,96 hours after culture.Cell morphology was compared by histopathological examination 3 weeks after culture.The mRNA expression of pigment epithelium-derived factor (PEDF),N-cadherin,β-catenin and cell connection related proteins in the cells of both groups were assayed using reverse transcription polymerase chain reaction (RT-PCR).Ultrastructure of the cells was observed under the transmission and scan electronic microscope 3 weeks after culture.Results The number of ARPE-19 cells cultured on denuded amniotic membrane was decreased significantly in comparison with the normal culture plate(F=41.760,P =0.000).Histopatholy also showed that the cell density on amniotic membrane was lower than of normal cells on plate surface.Moreover,the expression level of claudin 1 mRNA,N-cadherin mRNA and PEDF mRNA were significantly up-regulated in denuded amniotic membrane group in comparison with control group (t=15.828,P=0.000 ;t=6.839,P=0.002 ;t=14.667,P=0.000),but the expression of Connexin 43 mRNA was down-regulated in denuded amniotic membrane group compared with control group(t=3.358,P=0.024).Ultrastructural examination revealed that ARPE-19 cells cultured on amniotic membrane exhibited a polygonal epithelial phenotype with cilium on the apical side,however,the cells cultured on normal culture plate displayed fusiform shape and uneven thickness.Conclusions Amniotic membrane plays a promoting effect on the differentiation of ARPE-19 cells and a inhibitory effect on the proliferation of ARPE-19 cells,suggesting that amniotic membrane might be an useful scaffold for the preparation of functionally mature RPE cells for clinical transplantation.

The epidemic of COVID-19 has brought great challenges to the global public health prevention and control system combined with clinical diagnosis and treatment system, and it makes the development of effective antiviral drugs an important task in current pharmaceutical research. Traditional Chinese medicine (TCM) has played an important role in the prevention and control of COVID-19. Due to its numerous chemical components and various structural types, TCM becomes a natural library for searching for lead compounds against SARS-CoV-2. In this study, a novel dual-target surface plasmon resonance (SPR) biosensor was developed for S protein receptor binding domain (SRBD) and angiotensin converting enzyme 2 (ACE2) which are two key proteins in the process of SARS-CoV-2 invading cells according to characteristics of synergistic effects of multiple components and comprehensive regulation of multiple targets of TCM. The SPR biosensor was applied to screen and identify active components from six TCMs, and daidzin from Puerariae Lobatae Radix was identified to bind with SRBD and ACE2. The affinity constant (K_D) of daidzin and ACE2 was 5.18 μmol·L^-1 through the SPR affinity assay. Competitive ELISA assay showed that daidzin could inhibit the binding of SRBD and ACE2, and the inhibition rate of daidzin (20 μmol·L^-1) was 38.6%. Molecular docking experiments further confirmed that daidzin had the best binding near the binding region of SRBD-ACE2 complex. This study shows that the dual-target SPR screening system is accurate and efficient, and is particularly suitable for screening of complex drug systems and effective substances study of TCM. It provides a material basis for exploring the mechanism of TCM active constituents against SARS-CoV-2, and provides a source of lead compounds for the development of anti-SARS-CoV-2 drugs.

AIM: To detect the differentiation effects of retinal cells or extracts on bone marrow-derived mesenchymal stem cells (BMSC).METHODS: Human fetal BMSC were previously labeled by carboxyfluorescein succinimidyl ester (CFSE), and co-cultured with retinal pigment epithelial （RPE） cells which were pre-treated with ultraviolet irradiation at a ratio of 1∶1 to induce the differentiation of BMSC for up to 14 days. In some assays, a retinal extract of bovine retinal extract (BRE) was added to detect the potential effects of retinal component on the differentiation of BMSC. In addition, neuron-specific enolase (NSE), Nestin and Glial fibrillary acidic protein (GFAP) immunostaining were performed to determine the characteristics of BMSC.RESULTS: The results indicated that by co-cultured with RPE cells, fetal BMSC were differentiated into neural-like cells expressing special neuronal markers Nestin, GFAP and NSE. And the expression of these markers was obviously increased by BRE.CONCLUSION: Retina derived cells and extracts can induce the differentiation of BMSC into neural-like cells.

Adolescent ; Adult ; Branchial Region ; abnormalities ; Child ; Female ; Fistula ; congenital ; surgery ; Humans ; Male ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Retrospective Studies ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the clinical effects of PLIF surgery for elderly patients with lumbar degenerative disease.

METHODSFrom March 2010 to May 2013, 28 patients with lumbar degenerative disease, aged more than 80 years were treated with PLIF surgery. There were 10 males and 18 females, aged from 80 to 93 years old with an average of (85.44±3.66) years. Course of disease was from 3 to 20 years. The operation time, intra-operative blood loss, operation complications were recorded and JOA scores and Macnab criteria were used to evaluate the clinical outcomes.

RESULTSAll patients were followed up from 12 to 40 months with an average of 26.5 months. The average operation time was (150.00±26.42) min and the average intra-operative blood loss was (373.33±99.88) ml. The pre-operative JOA score was 12.30±2.43, and the corresponding postoperative JOA score at the final follow-up was 24.81±2.09 which was much higher than the preoperative one (P<0.01). According to the modified Macnab criteria to evaluate at the final follow-up, 16 patients got an excellent result, 10 good, 2 fair. In the weeks postoperatively, injuries of nerve root happened in 3 cases, superficial wound infection with delayed healing in 3 cases, and tear of the dural sac accompanied with cerebrospinal fluid leakage in 1 case. After long term follow-up, adjacent segment degeneration and the corresponding spinal canal stenosis occurred in 1 case at 34 months after operation. All cases got successful fusion without any displacement of internal fixation and pseudoarthrosis formation.

CONCLUSIONWith proper cases, fully preoperative preparation, perfect intra-operative manipulation and active treatment after operation, even advanced ages older than 80 years with lumbar degenerative disease could get satisfactory outcomes after PLIF surgery.

Aged ; Aged, 80 and over ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Operative Time ; Spinal Diseases ; surgery ; Spinal Fusion ; methods

OBJECTIVETo explore the clinical effects of transforaminal endoscopic spine system in surgical revision of lumbar vertebrae.

METHODSFrom January 2012 to October 2013,14 patients who needed reoperations of lumbar vertebrae were treated using transforaminal endoscopic spine system (TESSYS). There were 8 males and 6 males, aged from 27 to 84 years old with an average of (50.4 ± 18.9) years. Visual analogue scale (VAS) and Japanese Orthopaedic Association Scores (JOA) were compared before and after surgical revision. Macnab standard was used to assess the clinical effect.

RESULTSAll the patients were followed up from 6 to 27 months with the mean of 18 months. Preoperative VAS score was 6.79 ± 1.31, and in a week,3 months and 6 months after operation were 2.50 ± 1.29, 2.21 ± 1.53, 1.64 ± 1.08, respectively, which were all much lower (P < 0.01) than preoperative score. Preoperative JOA score was 12.43 ± 1.95, and the above corresponding postoperative JOA scores were 21.50 ± 3.78, 21.93 ± 4.55, 23.36 ± 4.33, respectively, which were all much higher than preoperative score (P < 0.01). According to the modified Macnab criteria, 5 patients got an excellent results, 7 good, 1 fair and 1 poor. The nerve root injury of L5 occurred in 1 case during paracentesis and no other complications were found.

CONCLUSIONSelecting the appropriate indications using TESSYS in surgical revision of lumbar vertebrae can successfully avoid the operation scar, reduce the surgical complications and obtain satisfactory clinical outcomes.

Adult ; Aged ; Aged, 80 and over ; Endoscopy ; methods ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Reoperation ; Spinal Fusion ; methods

Objective To construct a lentiviral vector carrying rat sirt1 gene and observe the expression of sirt1 in retinal ganglion cell (RGC) of rat.Methods Rat sirt1 cDNA was inserted into pLV5 vector.After identification by sequencing analysis and PCR,the recombinant sirt1expressinglentivirus vector was packaged by cotransfecting 293T cells with packaged plasmid.Then pLV5-sirt1 was used to infect the cultured Sprague-Dawley rat RGC cell in vitro.The expressions of sirt1 protein and mRNA in infected rat RGC were detected by quantitative real-time PCR and Western blot.Results The sirt1 expression vector pLV5 was successful constructed and sequence was proved to be correct.The expression of sirt1 protein and mRNA in RGC was significantly increased than that in cells infected with control lentiviruses (P ＜ 0.05).Conclusion We have successful constructed a sirt1 expression lentivirus vector pLV5-sirt1 and it can increase the expression of sirt1 protein and mRNA in the rat retinal ganglion cells.