AIM: To investigate whether grafting neural stem cells (NSCs) improves the impaired cognitive deficits and spatial recognition after ischemic-hypoxic brain damage (HIBD) in neonatal rats. METHODS: Non-immunosuppressed 7-day-old SD rats were used as research subject and randomly divided into 3 groups: (1) sham group (n=10); (2) HIBD group (n=11); (3) transplant group (n=13). (2) and (3) were anesthetized and subjected to a hypoxic/ischemic injury obtained by combination of left carotid ligation and exposure to 8% oxygen for 2 h. At 3 days post injury, hypoxic-ischemic brain damaged animals were re-anesthetized and randomized to receive stereotactic injection of NSCs prelabeling with BrdU or control media into the hippocampus in the ipsilateral hemisphere. Cognitive (i.e., learning) deficits were assessed at 2 to 4 weeks after transplantation. At the end of the behavioral tests, the animals were killed and evaluated for NSC survival and histopathological analysis. RESULTS: Transplant group showed significantly improved cognitive function in selected tests as compared with HIBD group during the 4-week observation period. They took less time than HIBD group in finding the 3 arms baited with water and had a decreased number of working and reference memory errors in radial maze acquisition tests. Histological analysis showed that transplanted NSCs attenuated CA1 cell loss after HIBD, and NSCs survived for as long as 4 weeks after transplantation and were detected in the hippocampus. CONCLUSION: These data suggest that transplanted NSCs attenuate brain damage and cognitive dysfunction after hypoxic-ischemic brain damage. This approach warrants continued investigation in light of potential therapeutic uses.

AIM: To discuss the mechanism of hyperbaric oxygen(HBO) therapy by assessing the changes of neural stem cells(NSCs),after hypoxic-ischemic brain damage(HIBD) in neonatal rats.METHODS: Seven-day-old SD rat pups were randomly divided into 4 groups: control group(CON,n=16),HIBD group(n=16),hyperbaric air group(HBA,n=16),and HBO group(n=16).The HIBD model was produced by permanent occlusion of left common carotid artery and was exposed to a mixture of 8% oxygen and 92% nitrogen for 2 h(at 37 ℃).HBA and HBO treatment was administered by placing pups in a chamber(2 ATA for 1 h) 1 h after hypoxia exposure and performed once daily for 7 days.BrdU immunohistochemistry was used to assess how the insult had affected NSCs in the SVZ of the lateral ventricle and DG of the hippocampus.The NSCs from the ipsilateral SVZs were isolated at 3 weeks recovery from hypoxia-ischemia(HI).The number of spheres was then counted as an index of the number of NSCs residing within the SVZ.RESULTS: At 3 week survival,the SVZ of HIBD group was smaller and markedly less cellular than control group.BrdU-positive cells were dramatically decreased in the SVZ and DG of the affected hemisphere(P

Objective To investigate the protective effects of hyperbaric oxygen(HBO)against brain dam- age from hypoxic ischemia(HIBD)in neonatal rats.Methods One hundred and seventeen 7-day-old Sprague- Dawley rats were randomly divided into 4 groups:a control group(n=32),a hypoxic ischemia brain damage group (HIBD group,n=30),a hyperbaric air group(HBA group,n=27),and a hyperbaric oxygen group(HBO group, n=28).The HIBD model was established by permanent occlusion of the left common carotid artery followed by expo- sure to a mixture of 8% oxygen/92% nitrogen for 2 h(at 37℃).HBO therapy was administered to the HBO group after the hypoxia exposure once a day for 7 d,as was HBA therapy to the HBA group.Apoptotic cells in the cortex and hippocampus(A_(CH)cells)were measured using TUNEL at 9 d after birth,and the ratios of left and right cerebral hemisphere weight(R_(L/R))and rate of weight gain(GRW)were recorded 14 d after birth.A radial arm maze acquisi- tion test(RAMAT)was administered at 30 to 35 days.Lastly,the neuron density in the CA_1 subfield of the rats' hip- pocampi(ND_(CAI)was measured with Nissl staining.Results R_(L/R)and GRW in the HIBD group were significantly lower than in the control group(P＜0.01),while R_(L/R)was increased in the HBO and HBA groups,especially in the HBO group(P＜0.01),although there was no significant difference in GRW between the groups.Compared with the control group,A_(CH)cells were increased and ND_(CAI)was decreased in the HIBD group(P＜0.01),while A_(CH)cells were decreased and ND_(CAI)was elevated in the HBO group in comparison with the HIBD group(P＜0.01).There was no change in A_(CH)cells or ND_(CAI)in the HBA group.The RAMAT results for the HIBD group,including the time to find the arms baited with water,average times of working errors and reference memory errors,were significantly high- er than those of the control group,while these values for the HBO group were obviously lower than for the HIBD group,and there was no change for the HBA group(P＞0.05).Conclusion HBO therapy might increase the re- covery of learning and memory function by attenuating HIBD in neonatal rats.

Objective:To evaluate the clinical outcome of botulinum A toxin(BTX-A)injection into external sphincter combined with oral baclofen in treatment of detrusor-external sphincter dyssynergia(DESD)after spinal cord injury(SCI). Methods:A total of 38 urodynamic examination-confirmed DESD patients,male 31 and female 7,with an average age of (36.5?17.8)years old,were included in this study.200 U of BTX-A toxin was dissolved in 8 ml of normal saline and the solution was injected at 8 different sites(1 ml per site)of the external sphincter via a 5F flexible cystoscopic needle.On the second day,9 patients(BTX-A+baclofen group)were randomly selected for baclofen oral administration,3/d for 3 months; the other 26 patients were taken as control.Urodynamic examination was repeated in all patients 4 weeks later;the voiding diary and urodynamic outcomes were compared before and after treatment.The adverse and toxic effects were observed in the patients who were followed up for 2-9 months.Results:One month after treatment the voiding and storing functions of bladder were improved to different degrees,with the mean maximum uroflow rate(Qmax),the mean urine volume,the mean maximal cystometric capacity and the bladder compliance increased significantly and the mean postvoid residual urine volume and the mean maximal voiding pressure decreased significantly(all P

Amino Acid Metabolism, Inborn Errors ; diagnosis ; metabolism ; therapy ; Diagnosis, Differential ; Diet ; Glutarates ; metabolism ; urine ; Humans ; Infant ; Male ; Riboflavin ; therapeutic use ; Treatment Outcome

Aim Neural injury in the central nervous system following is chemic insult is believed to result from oxygen and glucose deprivation.In this study,baicalin was investigated for its neuroprotective effects against oxygen and glucose deprivation(OGD) in Neuro2A cells.Methods Mitochondrial activity was assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide(MTT) reduction activity assay.Apoptosis was monitored with flow cytometry.It was found that baicalin increased MTT reduction activity and decreased percentage of apoptosis of the cultured Neuro2A cells.Results Baicalin showed significant protective effects on the OGD-induced apoptosis in cultured Neuro2A cells.Conclusion These results suggest that baicalin is an effective compound in preventing neurotoxicity induced by OGD and therefore deserves further scrutiny.

AIM: To investigate the influence and mechanism of blue light on the proliferation of human retinal pigment epithelial cells.METHODS: Cells were divided into two groups,including blue light group and control group.The 35W white light lamp with blue filter was used to establish damaged RPE cell model in vitro.Blue ray wavelength ranged between 470nm and 520nm.And the light intensity was about 2000Lx.After exposure to blue light,we tested the proliferation of human retinal pigment epithelial cells by CCK-8 kit.And then expression of miR-103 was measured by the real-time PCR.RESULTS: Exposure to blue light inhibited the proliferation of human retinal pigment epithelial cells and increased the expression of miR-103.Moreover,up-regulation of miR-103 inhibited the proliferation of human retinal pigment epithelial cells,and down-regulates miR-103 promoted the proliferation of human retinal pigment epithelial cells.CONCLUSION: Blue light inhibits the proliferation of human retinal pigment epithelial cells by the up-regulation of miR-103.

To construct a recombinant expression system for a single-domain antibody targeting the urease of Helicobacter pylori (Hp), this study employed several strategies. First, using artificial intelligence (AI) auxiliary tools such as Pymol, I-TASSER, and ClussPro2, the molecular interactions between different antibodies and Hp urease subunit B (UreB) were analyzed. The fully humanized single-domain antibody UreBAb was identified as the primary research target. Next, the UreBAb gene sequence was optimized based on Escherichia coli codon preferences, and was inserted into expression vectors such as pET28a and pE-SUMO. The resulting recombinant expression strains were obtained by transforming Escherichia coli Rosetta(DE3). Recombinant antibody proteins were prepared through IPTG induction, and its activity was detected using extracted Hp urease as the antigen. SDS-PAGE analysis confirmed the correct expression of both UreBAb and SUMO-UreBAb, with protein yields of 0.34 mg/mL and 0.41 mg/mL, respectively. Unidirectional immunodiffusion experiments further confirmed that both recombinant antibodies exhibited strong affinity for Hp UreB antigen, with inhibition rates of 51.27% and 74.07%, respectively. Additionally, leveraging artificial intelligence tools such as AlphaFold2, cluspro2, mCSM-AB, OSPREY, and FoldX, the study evaluated and analyzed key binding sites and mutational strategies affecting the stability of the antigen-antibody complex. Subsequently, nine UreBAb evolution mutants were constructed, and their binding activities with the antigen were enhanced. Among these, the I107W mutant showed the most significant improvement, achieving a 24.95% increase compared to the wild-type UreBAb. This research lays a solid foundation for the development of fully humanized single-domain antibodies against Hp.

OBJECTIVE: To explore the effect of apoB polymorphism on plasma lipid levels and cerebral hemorrhage in (CH) Changsha Han Chinese. METHODS: One hundred thirty CH patients and 100 normal people were involved. C7673T polymorphism of apoprotein B was analyzed by PCR-restriction fragment length polymorphism (PCR-PFLP); and the triglyceride(TG), total cholesterol (TC), high density lipoprotein-cholesterol (HDL), and low density lipoprotein-cholesterol(LDL) levels were examined by oxidase method. The serum level of lipoprotein(a) was determined by immunology method. RESULTS: (1) Allele T frequencies of apoB C7673T in CH patients and the control group were 0.108 and 0.040, respectively. Allele T frequencies of apoB C7673T in the CH patients were significantly higher than those in the control group (P< 0.01). (2) In the CH patients, the levels of TC and LDLjC of the T/C gene type were significantly higher than those of the C/C gene type, while the levels of HDLjC of the T/C gene type were significantly lower than those of the C/C gene type (P< 0.05). CONCLUSION ApoB C7673T polymorphism may be related to cerebral hemorrhage, and the changing blood lipid level may increase the susceptibility of CH.
Aged ; Apolipoproteins B ; genetics ; Cerebral Hemorrhage ; blood ; genetics ; China ; ethnology ; Female ; Genetic Predisposition to Disease ; Humans ; Lipoproteins ; blood ; Male ; Middle Aged ; Polymorphism, Genetic ; genetics

OBJECTIVETo investigate the impact of Staphylococcus aureus from infertile men on sperm motility and the relationship between virulence genes and the activity of spermatozoal immobilization.

METHODSWe collected 60 strains of non-repeated Staphylococcus aureus from the semen of 589 infertile males and analyzed the influence of Staphylococcus aureus on sperm motility using the computer-aided sperm analysis system. We selected the strains that apparently decreased sperm motility and detected their virulence genes by PCR.

RESULTSSperm motility was significantly decreased in 17 of the 60 strains of Staphylococcus aureus (P < 0.05). The main virulence genes in these strains were hlg (33.3%), scn (23.3%), cna (20%), hlb (20%), and clfA (18.3%), others including icaA, fnbA, tst, seb, hld, eta and sea. The scn gene carriers accounted for 47.1% in the spermatozal immobilization positive group, significantly higher than 14% in the negative group (P < 0.05). No statistically significant differences were found in the percentages of the carriers of the other virulence genes between the two groups (P > 0.05).

CONCLUSIONInfections of Staphylococcus aureus in male reproductive system can lead to the decrease of sperm motility, which may be associated with the Staphylococcus complement inhibitor encoding gene scn.

Humans ; Infertility, Male ; microbiology ; Male ; Polymerase Chain Reaction ; Semen ; microbiology ; Species Specificity ; Sperm Motility ; Staphylococcal Infections ; Staphylococcus aureus ; pathogenicity ; Virulence ; genetics