Objective To investigate the migration and survival of neural stem cells(NSCs)in vivo.Methods NSCs cultured in vitro were transfected by lentiviral vectors expressing green fluorescent protein(GFP)to construct GFP-NSCs,then trans-seeded into lactide-co-glycolide(PLGA)scaffold and implanted into the injured site of T9 spinal cord in rat.One month after transplantation,the migration of NSCs in spinal cord was examined by fluorescence microscope,and the survival rate of NSCs was counted out.Results NSCs labeled GFP had strong expression of green fluorescence.One month after transplanting,part of NSCs expressing GFP could be seen in PLGA scaffolds and rostral,caudal spinal core.The survival rate counted out was 1.4911±0.0313%.Conclusion NSCs marked by GFP and transplanted to rat injured spinal cord could migrate into the spinal cord tissues and the minority of them could survive.

Objective To explore the effect of transplanting poly (lactic-co-glycolic acid) (PLGA) scaffolds seeded with neural stemcells (NSCs) and Schwann cells (SCs) on spinal cord injured rats and the mechanism. Methods NSCs and SCs were cultured in vitro andthen seeded into the directional PLGA scaffolds. Then PLGA-cell complexes were implanted into the spinal cord hemisected rats, whichwere divided into PLGA group, PLGA+NSCs group and PLGA+ NSC+SCs group. The rats were tested with cortical motor evoked potentials(CMEPs) and Basso-Beattle-Bresnahan (BBB) score. Then, the rats were further ipsilaterally or contralateral hemisected at T6 and testedwith CMEPs and BBB score again. Results The incidence of recovery and the amplitudes of CMEPs were the highest in PLGA+NSCs+SCs group. The rats exhibited a gradual improvement in hindlimb locomotor function in score. The BBB score was the least in the PLGAgroup in the 2nd week or later. After retransected ipsilaterally, the CMEPs disappeared again and the BBB score improved quickly. But afterretransected contralaterally, the rats were completely paraplegia. Conclusion The directional PLGA scaffolds seeded with NSCs and SCs facilitatethe recovery in spinal cord injured rats, which may associate with axonal regeneration and functional connections, but play a limitedrole.

AIM: To investigate the effects of astragalus injection combined with puerarin injection on endo-plasmic reticulum stress through PERK pathway in diabetic nephropathy mice .METHODS: Male KKAy mice were ran-domly divided into model group ( injected with normal saline ) and treatment group ( injected with astragalus and puerarin ) . The male C57BL/6J mice served as normal group .The mice were sacrificed 4 weeks after treatments for observing morpho-logical changes under electron microscope .The renal tissues were collected to determine the expression of protein kinase R-like endoplasmic reticulum kinase ( PERK ) , eukaryotic initiation factor 2α( eIF2α) and glucose-regulated protein 78 (GRP78) at mRNA and protein levels by real-time PCR and Western blot.RESULTS: Under electron microscope, the renal tubular epithelial cells in model group and treatment group showed the swelling of the nucleus , endoplasmic reticulum and mitochondria .The results of real-time PCR and Western blot showed that the expression of PERK , eIF2αand GRP78 at mRNA and protein levels in model group was higher than that in normal group (P<0.05), while that in treatment group was lower than that in model group .CONCLUSION: Astragalus injection combined with puerarin injection reduces the mRNA and protein expression of PERK , eIF2αand GRP78, thus inhibiting the endoplasmic reticulum stress in type 2 dia-betic mice to protect the kidney function .

Objective:To explore vibration, position and motion proprioception of the ankle joints after a stroke.Methods:Twenty-eight stroke survivors with impaired ankle proprioception were divided into a right-side stroke group ( n=18) and a left-side stroke group ( n=8). Twenty-two healthy volunteers constituted a control group. Vibration perception thresholds, passive and active joint angle resetting, and motion minimum thresholds were quantified among the stroke survivors on both the healthy and the affected side. With the controls the dominant and non-dominant sides were used. The differences in proprioception between the healthy volunteers and the stroke patients, between the affected side and the healthy side of the stroke patients, and between left- and right-side stroke patients were analyzed and compared. Results:Among the stroke survivors the vibration perception threshold on the affected side averaged (28.91±22.53)μm. The absolute difference in the perception of passive positioning was (5.49±5.39)° for 15° of plantar flexion and (4.48±3.89)° for 5° of dorsal extension. In active positioning plantar flexion was (5.23±4.34)° and for 30° of plantar flexion it was (3.26±1.73)°. The 5° dorsal extension error was (4.97±3.48)°. The motion perception thresholds between 20° of plantar flexion, 10° of plantar flexion and the neutral position were significantly higher, on average, than among the control group. The stroke group also had significantly higher motion perception thresholds than the control group.Conclusion:The vibration, position, and motion sense of the ankle joint on a stroke survivor′s affected side tend to be impaired, with the impairment of vibration and motion sensing tend to be more substantial. After stroke, there is also mild impairment of vibration, position and motion sensing in the healthy ankle joint. The impairment of proprioception caused by right cerebral hemisphere injury may be more serious than that caused by injury on the left.

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj14-Sj26 that contains fatty binding protein (Sj14), GST (Sj26) and paramyocin (Sj97) that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was trans-fected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and pIRES-Sj97-Sj14-Sj26 plasmid DNA, pIRES-Sj14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA,plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh.Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the pIRES-Sj97-Sj14-Sj26 group as compared with the plRES blank vector, normal saline and pIRES-Sj26 groups (P<0.01) and the pIRES-Sj14-Sj26(P<0.05). Single splenocyte suspension was prepared to detected the level of IFN-γ by ELISA and the lymphocyte stimulating index (SI) by MTT SI was significantly higher of in the pIRES-Sj97-Sj14-Sj26 group than in other groups (P<0.01), while the IFN-γ, level was significantly higher the pIRES-Sj97-Sj14-Sj26 group than in pIRES blank vector and normal saline groups (P<0.01), but no significant differences were found when compared with pIRES-Sj14-Sj26 and pIRES-Sj26 groups. Flow cytometery showed that the percent-ages of CD4+ and CD8+ T cells were much higher in the pIRES-Sj97-Sj14-Sj26 group (P<0.01,P<0.05). It was concluded that pIRES-Sj97-Sj14-Sj26 vaccine may induce stronger immune response in BALB/c mice.

When the coverage of the vaccinated people reaches a certain percentage of the population, the herd protection will protect the unvaccinated persons. However, the traditional clinical evaluation of vaccines performing individual randomized design fails to evaluate the herd protection of vaccines. Compared with the individual randomized design, the cluster-randomized design can determine the overall protection by the vaccine more comprehensively. It has become increasingly common to perform a cluster-randomized design in clinical trials of vaccines in Phase Ⅲ and Ⅳ clinical trials. However, little is known about the application of cluster randomized design in vaccine clinical trials in China. We, at this moment, do a review of the application of random cluster design in vaccine clinical trials and provide references for future research in China.

OBJECTIVE: To investigate the effect of short-term rehabilitation therapy based on exercise on lung function in coal workers' with pneumoconiosis(CWP). METHODS: A total of 74 CWP patients were divided into control group(32) and treatment group(42) by random number table method. The control group received routine treatment only. The treatment group underwent 6 months of exercise-based rehabilitation treatment on the basis of routine treatment. The lung function was assessed in two groups to evaluate the treatment efficacy. RESULTS: Before rehabilitation treatment, the vital capacity(VC) and forced vital capacity(FVC) of patients in the treatment group were lower than that in the control group(P<0.05). But there was no significant difference in forced expiratory volume in first second(FEV_(1.0)) and FEV_(1.0)% between the two groups(P>0.05). After treatment, VC and FVC in the treatment group were higher than that before treatment in the same group(P<0.05). However, there was no significant difference in the four lung function indexes before and after treatment in the control group(P>0.05). The difference of VC and FVC before and after treatment in the treatment group was higher than those in the control group(P<0.01). However, there was no significant difference in FEV_(1.0 )and FEV_(1.0)% between the two groups before and after treatment(P>0.05). CONCLUSION: Exercise-based short-term rehabilitation therapy can improve lung ventilation of CWP patients.

Objective:To investigate the impacts and mechanisms of the GCH1-S81A mutants of the rate-limiting enzyme GTP cyclohydrolase 1 (GCH1) at key phosphorylation sites on the radiosensitivity of esophageal cancers during the de novo synthesis of tetrahydrobiopterin (BH4). Methods:The KYSE-150 cell lines of esophageal squamous cell carcinoma with stable GCH1 overexpression at different phosphorylation levels were constructed in this study. Based on GCH1′s phosphorylation levels and radiation doses, the experimental groups were divided into the blank control group, the empty virus group, the empty virus + irradiation group, the wild-type GCH1 group, the GCH1 phosphorylation group, the GCH1 dephosphorylation group, the GCH1 dephosphorylation + irradiation group, the validation group, and the validation + irradiation group. The Western blot and the CCK-8 assay were employed to detect the infection efficiency and the survival rates of tumor cells in various groups, respectively. The flow cytometry was applied to detect the changes in the apoptosis rate, reactive oxygen species (ROS) level, and lipid peroxide level of tumor cells in various groups. The colony formation assay was used to detect the changes in the radiosensitivity of tumor cells. Besides, the Western blot was performed to detect the changes in the expression of ferroptosis-related proteins.Results:The GCH1 dephosphorylation group showed a significantly decreased expression level of phosphorylated GCH1-S81 protein in the cells at 48 h after infection ( t=9.35, 16.57, P<0.05). Compared to the empty virus + irradiation group, the GCH1 dephosphorylation + irradiation group exhibited a significantly decreased cell survival rate 24 h after 10 Gy X-ray irradiation ( t=26.97, P<0.05). The ROS levels in KYSE-150 cells at 8 h after 10 Gy X-ray irradiation, and the apoptosis rates and lipid peroxide levels at 48 h after irradiation, all showed a significant increase ( t=17.89-131.1, P<0.05), which was further aggravated following the addition of GCH1-S81A mutants ( t=157.06-97.81, P<0.05). This phenomenon could be inhibited by complementing wild-type GCH1 ( t=66.38-23.08, P<0.05). Compared to the empty virus + irradiation group, the GCH1 dephosphorylation + irradiation group manifested decreased colony formation capacity under various X-ray doses (2, 4, 6 and 8 Gy, t=7.31-8.16, P<0.05). The GCH1-S81A mutation reduced the expression level of the ferroptosis-related protein GPX4 ( t=4.55, P<0.05), which was further decreased after 10 Gy X-ray irradiation ( t=12.98, P<0.05). Conclusions:The GCH1-S81A dephosphorylated mutants can inhibit the growth of esophageal carcinoma cells KYSE-150 and enhance their radiosensitivity, which may hold promise as a novel approach to improve the efficacy of radiotherapy for esophageal cancers.

Chronic myeloid leukemia (CML) is a slowly progressing hematopoietic cell disorder. Sphingosine kinase 1 (SPHK1) plays established roles in tumor initiation, progression, and chemotherapy resistance in a wide range of cancers, including leukemia.However, small-molecule inhibitors targeting SPHK1 in CML still need to be developed. This study revealed the role of SPHK1 in CML and investigated the potential anti-leukemic activity of hirsuteine (HST), an indole alkaloid obtained from the oriental plant Uncaria rhynchophylla, in CML cells. These results suggest that SPHK1 is highly expressed in CML cells and that overexpression of SPHK1 represents poor clinical outcomes in CML patients. HST exposure led to G2/M phase arrest, cellular apoptosis, and downregulation of Cyclin B1 and CDC2 and cleavage of Caspase 3 and PARP in CML cells. HST shifted sphingolipid rheostat from sphingosine 1-phosphate (S1P) towards the ceramide coupled with a marked inhibition of SPHK1. Mechanistically, HST significantly blocked SPHK1/S1P/S1PR1 and BCR-ABL/PI3K/Akt pathways. In addition, HST can be docked with residues of SPHK1 and shifts the SPHK1 melting curve, indicating the potential protein-ligand interactions between SPHK1 and HST in both CML cells. SPHK1 overexpression impaired apoptosis and proliferation of CML cells induced by HST alone. These results suggest that HST, which may serve as a novel and specific SPHK1 inhibitor, exerts anti-leukemic activity by inhibiting the SPHK1/S1P/ S1PR1 and BCR-ABL/PI3K/Akt pathways in CML cells, thus conferring HST as a promising anti-leukemic drug for CML therapy in the future.

Objective:To analyze the accuracy of lung ultrasound and chest X-ray in the diagnosis of neonatal pulmonary disease.Methods:We prospectively collected newborns that needed chest X-ray examination to diagnose pulmonary disease from twelve neonatal intensive care units across the country between June 2019 and April 2020.Each newborn was examined by lung ultrasound within two hours after chest X-ray examination.All chest X-ray and lung ultrasound images were independently read by a radiologist and a sonographer.When there was a disagreement, a panel of two experienced physicians made a final diagnosis based on the clinical history, chest X-ray and lung ultrasound images.Results:A total of 1 100 newborns were enrolled in our study.The diagnostic agreement between chest X-ray and lung ultrasound(Cohen′s kappa coefficient=0.347) was fair.Lung ultrasound(area under the curve=0.778; 95% CI 0.753-0.803) performed significantly better than chest X-ray(area under the curve=0.513; 95% CI 0.483-0.543) in the diagnosis of transient tachypnea of the newborn( P<0.001). The accuracy of lung ultrasound in diagnosing neonatal respiratory distress syndrome, meconium aspiration syndrome, pneumonia and neonatal pulmonary atelectasis was similar to that of chest X-ray. Conclusion:Lung ultrasound, as a low-cost, simple and radiation-free auxiliary examination method, has a diagnostic accuracy close to or even better than that of chest X-ray, which may replace chest X-ray in the diagnosis of some neonatal lung diseases.It should be noted that both chest X-ray and lung ultrasound can only be used as auxiliary means for the diagnosis of lung diseases, and it is necessary to combine imaging with the clinical history and presentation.