Construction, screening and immunogenicity of the recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 / 细胞与分子免疫学杂志

Objective To construct a recombinant poxvirus vector vaccine, rVTTδTK-RBD, and to evaluate its safety and immunogenicity. Methods The receptor-binding domain (RBD) gene was synthesized with reference to the gene sequence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and was inserted into the polyclonal site of the self-constructed recombinant plasmid pSTKE, to construct the recombinant poxvirus shuttle vector pSTKE-RBD. This was then transfected into BHK-21 cells pre-infected with the vaccinia virus Tiantan strain (VTT). The recombinant poxvirus rVTTδTK-RBD was successfully obtained after several rounds of fluorescence phage screening. The effect of rVTTδTK-RBD on the body mass of BALB/c mice was detected after immunizing mice by intra-nasal vaccination. The levels of specific and neutralizing antibodies produced by rVTTδTK-RBD on BALB/c mice were analyzed after immunizing mice intramuscularly. The effect of rVTTδTK-RBD on T cell subsets in BALB/c mice was detected by flow cytometry. Results Through homologous recombination, enhanced green fluorescent protein (EGFP) screening marker, and multiple rounds of fluorescent phosphorescence phage screening, a recombinant poxvirus rVTTδTK-RBD, expressing RBD with deletions in the thymidine kinase (TK) gene, was successfully obtained, which was validated by PCR. The in vivo experiments on BALB/c mice showed that rVTTδTK-RBD was highly immunogenic against SARS-CoV-2 and significantly reduced toxicity to the body compared to the parental strain VTT. Conclusion The recombinant poxvirus vaccine rVTTδTK-RBD against SARS-CoV-2 is successfully constructed and obtained, with its safety and immunogenicity confirmed through various experiments.

Evaluating the application efficiency of the MHSeqTyper47 kit in kinship identification / 中国法医学杂志

Objective To investigate the application value of the MHSeqTyper47 kit in kinship identification.Methods Multiplexed amplification and library preparation were performed for DNA samples from 113 related individuals by using the MHSeqTyper47 kit.The libraries were sequenced on a MiSeq FGx sequencer,and the data were analyzed using MHTyper for microhaplotype genotyping.The kinship indexes were calculated to evaluate the application efficiency of this kit in kinship identification and compared with those of the GlobalFilerTM kit.Results For the MHSeqTyper47 kit,the CPI values in trio identification were 1.43× 1011～6.15×1018.The CPI values in duo identification were 1.02× 105～1.53× 1013.The CFSI values in full sibling identification were 7.73×101～2.59×1016.Trios,duos and full siblings could be completely distinguished from unrelated pairs.The combined efficiency of these two kits in 2nd-degree kinship identification was 0.466 2.Conclusion The application value of MHSeqTyper47 kit is relatively higher in the identification of lst-degree kinships.If jointly used with the GlobalFilerrM kit,2nd-degree kinship identification could be achieved in some cases.

Different calcium ion concentrations affect epithelial mesenchymal transformation of human peritoneal mesothelial cells via endoplasmic reticulum stress / 中华危重病急救医学

Objective:To study the effects of different calcium ion concentrations on epithelial mesenchymal transformation (EMT) of human peritoneal mesothelial cell (HPMC) via endoplasmic reticulum stress (ERS).Methods:HPMC cell line HMrSV5 was cultured in vitro and treated in groups. The cells in the control group, high calcium group 1, and high calcium group 2 were treated with medium containing calcium ion concentrations of 1.25, 1.75, and 2.25 mmol/L, respectively. The solvent control group was treated with medium containing 1.25 mmol/L physiological calcium ion concentration and 0.1% dimethyl sulfoxide (DMSO), the high calcium+solvent group was treated with medium containing 2.25 mmol/L calcium ion concentration and 0.1% DMSO, the high calcium+4-phenylbutyric acid (4-PBA) group was treated with medium containing 2.25 mmol/L calcium ion concentration and 1 mmol/L ERS inhibitor 4-PBA, and each group was treated for 48 hours. Morphological changes of cells in each group were observed under light microscope. The expressions of epithelial cell phenotype marker zonula occluden-1 (ZO-1) and mesenchymal cell phenotype marker α-smooth muscle actin (α-SMA) in the cells were observed by immunofluorescence staining. The expressions of EMT marker genes E-cadherin, ZO-1, α-SMA and Vimentin were detected by fluorescence quantitative polymerase chain reaction (PCR). The expressions of ERS marker proteins phosphorylated protein kinase R-like endoplasmic reticulum kinase (p-PERK), phosphorylated eukaryotic initiation factor 2α (p-eIF2α), transcription activating factor 4 (ATF4) and C/EBP homologous protein (CHOP) were detected by Western blotting. Results:Compared with the control group, the morphology of HMrSV5 cells became slender and fibrotic, the fluorescence intensity of ZO-1 increased, and the fluorescence intensity of α-SMA decreased in high calcium 1 and high calcium 2 groups, indicating that the cells transformed from epithelial cells to mesenchyme cells. The mRNA expressions of E-cadherin and ZO-1 were significantly decreased, while the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly increased, moreover, the expressions of the above marker genes or proteins in the high calcium 2 group was more obvious than those in the high calcium 1 group [E-cadherin mRNA (2 -ΔΔCt): 0.53±0.05 vs. 0.75±0.09, ZO-1 mRNA (2 -ΔΔCt): 0.42±0.06 vs. 0.69±0.06, α-SMA mRNA (2 -ΔΔCt): 1.81±0.16 vs. 1.32±0.14, Vimentin mRNA (2 -ΔΔCt): 2.05±0.22 vs. 1.48±0.16, p-PERK protein (p-PERK/β-actin): 0.81±0.09 vs. 0.59±0.06, p-eIF2α protein (p-eIF2α/β-actin): 0.87±0.10 vs. 0.50±0.06, ATF4 protein (ATF4/β-actin): 0.93±0.10 vs. 0.72±0.06, CHOP protein (CHOP/β-actin): 0.79±0.09 vs. 0.46±0.04, all P < 0.05]. Compared with the solvent control group, the morphological changes of cells, the expressions of EMT marker genes and ERS marker proteins after high calcium ion concentration of 2.25 mmol/L were consistent with those in the high calcium 2 group than control group. Compared with the high calcium+solvent group, the cell morphology recovered the characteristics of polygonal and pebble-like epithelial cells in the high calcium+4-PBA group, the fluorescence intensity of ZO-1 increased, the fluorescence intensity of α-SMA decreased, and the mRNA expressions of E-cadherin and ZO-1 in the cells were significantly increased [E-cadherin mRNA (2 -ΔΔCt): 0.86±0.09 vs. 0.57±0.04, ZO-1 mRNA (2 -ΔΔCt): 0.81±0.06 vs. 0.48±0.05, both P < 0.05], the mRNA expressions of α-SMA and Vimentin and the protein expressions of p-PERK, p-eIF2α, ATF4 and CHOP were significantly decreased [α-SMA mRNA (2 -ΔΔCt): 1.21±0.13 vs. 1.77±0.15, Vimentin mRNA (2 -ΔΔCt): 1.30±0.14 vs. 1.94±0.20, p-PERK protein (p-PERK/β-actin): 0.38±0.04 vs. 0.92±0.11, p-eIF2α protein (p-eIF2α/β-actin): 0.34±0.05 vs. 1.05±0.13, ATF4 protein (ATF4/β-actin): 0.57±0.06 vs. 0.97±0.11, CHOP protein (CHOP/β-actin): 0.51±0.04 vs. 0.90±0.12, all P < 0.05]. Conclusion:High calcium ion concentrations of 1.75 mmol/L and 2.25 mmol/L promote EMT of HPMC via activating ERS.

Effect of respiratory training based on core stability training on feedforward control in patients with chronic nonspecific low back pain / 中国康复理论与实践

ObjectiveTo investigate the effect of respiratory training based on core stability training on feedforward control in patients with chronic nonspecific low back pain (CNLBP). MethodsA total of 60 patients with CNLBP in Jiaxing Second Hospital from January, 2022 to March, 2023 were randomly divided into control group (n = 30) and experimental group (n = 30). Both groups received health education, physical factor therapy and core stability training, while the experimental group received respiratory training in addition, for four weeks. Visual Analogue Scale (VAS) score, Japanese Orthopaedic Association low back pain (JOA) score and Oswestry Dysfunction Index (ODI) were compared between two groups before and after treatment, while surface electromyography was used to detect the root mean square (RMS) and integrated electromyography (iEMG) of transversus abdominis, multifidus and triceps (movement muscles), and the activation sequence and relative activation time of transversus abdominis, multifidus and triceps were calculated. ResultsAfter treatment, the scores of VAS, JOA and ODI improved significantly in both groups (|t| > 8.515, P < 0.001), and the scores were better in the experimental group than in the control group (|t| > 2.089, P < 0.05). RMS and iEMG of transversus abdominis and multifidus improved significantly after treatment in both groups (|t| > 18.831, P < 0.001), and were significantly better in the experimental group (|t| > 3.481, P < 0.05). The transversus abdominis and multifidus in both groups were activated before the movement muscles, and the relative activation time of transversus abdominis and multifidus increased in negative (|t| > 48.115, P < 0.001), the experimental group being better (|t| > 3.229, P < 0.05). ConclusionCombination of core stability training and respiratory training is beneficial in reducing the pain of patients with CNLBP, reducing the lumbar dysfunction, improving the order of muscle activation, and strengthening feedforward control.

The Characteristics of Auditory Brainstem Response Waveform in Autistic Children with Normal Hearing / 听力学及言语疾病杂志

Objective To investigate the effect of auditory brainstem response(ABR)in clinical detection and severity assessment of autism spectrum disorder(ASD)in children with normal hearing.Methods ① A total of 55 autistic children(110 ears)with normal hearing and 55 children(110 ears)with typical development(TD)who did not differ in sex composition ratio and average monthly age were divided into four sub-groups according to age:≤24 months group(22 ears),25～36 months group(40 ears),37～48 months group(28 ears)and＞48 months group(20 ears).The ABR latencies and interpeak latencies were compared between ASD children and age-matched TD children.② ASD children were graded by severity according to the Diagnostic and Statistical Manual of Mental Dis-orders(DSM-V),and the correlations between the ABR latencies and interpeak latencies in autistic children with normal hearing and the severity grading were studied.Results ① No statistically significant differences in ABR wave latencies and interpeak latencies were found in autistic children with normal hearing under 24 months of age compared to age-matched TD children(P＞0.05).② Compared with children with TD,autistic children with nor-mal hearing at 25～36 months of age had significantly longer wave Ⅲ latencies and the interpeak latencies of Ⅰ-Ⅲ andⅠ-Ⅴ;the significantly longer wave Ⅲ,Ⅴ latencies,the interpeak latencies of Ⅰ-Ⅲ,Ⅲ-Ⅴ and Ⅰ-Ⅴ in autistic chil-dren with normal hearing at 37～48 months of age.Autistic children with normal hearing in the＞48 months group had significantly longer wave Ⅴ latencies and interpeak latencies of Ⅲ-Ⅴ,Ⅰ-Ⅴ than age-matched TD children(P＜0.05).③ The higher the ASD severity grading the longer the wave Ⅲ and V latencies and the longer interpeak latencies of Ⅰ-Ⅲ,Ⅲ-Ⅴ,and Ⅰ-Ⅴ(P＜0.05).Conclusion Differences in the level of auditory brainstem pathway de-velopment emerged at 25 months of age,and autistic children with normal hearing had significantly lower levels of auditory brainstem development than age-matched TD children.There were correlations between the latencies and interpeak latencies of ABR in autistic children with normal hearing and the severity grading.

Preliminary study of small intestinal manometry on gastrointestinal motility disorders / 中国医师进修杂志

Objective:To investigate the significance of small intestinal manometry in clinical diagnosis and treatment of patients with gastrointestinal motility disorders.Methods:The clinical data of 20 suspected patients with gastrointestinal motility disorders from February 2019 to January 2020 in Shanghai Jiaotong University Affiliated Ninth People′s Hospital were retrospectively analyzed. All patients were performed small intestinal manometry, and the influence on diagnosis and treatment as well as the clinical value of this technique were analyzed.Results:Among the 20 patients, 18 patients successfully accepted small intestinal manometry. Among the 18 patients, 14 patients showed significant motility abnormalities, among which 9 patients showed significant nutritional improvement after treatment and 5 patients were treated with maintaining nutritional support. In 10 patients, small intestinal manometry served as a deciding diagnostic tool, and in 12 patients, major modifications of treatment strategy were made according to results of small intestinal manometry.Conclusions:The small intestinal manometry significantly improves diagnosis and treatment decision in the patients with gastrointestinal motility disorders and shows great value on the clinical practice on this group of patients.

Analysis of 13 cases of enteric dysmotility / 中国医师进修杂志

Objective:To investigate the clinical characters of enteric dysmotility (ED), in order to improve clinical diagnosis and treatment capabilities.Methods:The clinical data of 13 ED patients underwent small intestinal manometry from August 2019 to July 2022 in the Shanghai Jiaotong University Affiliated Ninth People′s Hospital and the Fourth Affiliated Hospital of Nanjing Medical University were retrospectively analyzed, including the clinical manifestation, small intestinal manometry result, treatment and prognosis.Results:Among 13 patients, 1 abnormality was presented in 9 cases, 2 abnormalities presented in 2 cases and 3 abnormalities presented in 2 cases. All 13 cases presented a history of constipation, 5 cases started with constipation and underwent subtotal colectomy. Three patients showed severe chronic abdominal pain with one of them opiate dependence. Eight patients underwent surgical treatment, all of which achieved nutritional improvement and symptom relief.Conclusions:ED is a concisely defined disease with clear diagnostic criteria. The surgery can increase the symptoms of some patients.

Chlorella sp.-ameliorated undesirable microenvironment promotes diabetic wound healing

Chronic diabetic wound remains a critical challenge suffering from the complicated negative microenvironments, such as high-glucose, excessive reactive oxygen species (ROS), hypoxia and malnutrition. Unfortunately, few strategies have been developed to ameliorate the multiple microenvironments simultaneously. In this study, Chlorella sp. (Chlorella) hydrogels were prepared against diabetic wounds. In vitro experiments demonstrated that living Chlorella could produce dissolved oxygen by photosynthesis, actively consume glucose and deplete ROS with the inherent antioxidants, during the daytime. At night, Chlorella was inactivated in situ by chlorine dioxide with human-body harmless concentration to utilize its abundant contents. It was verified in vitro that the inactivated-Chlorella could supply nutrition, relieve inflammation and terminate the oxygen-consumption of Chlorella-respiration. The advantages of living Chlorella and its contents were integrated ingeniously. The abovementioned functions were proven to accelerate cell proliferation, migration and angiogenesis in vitro. Then, streptozotocin-induced diabetic mice were employed for further validation. The in vivo outcomes confirmed that Chlorella could ameliorate the undesirable microenvironments, including hypoxia, high-glucose, excessive-ROS and chronic inflammation, thereby synergistically promoting tissue regeneration. Given the results above, Chlorella is considered as a tailor-made therapeutic strategy for diabetic wound healing.

Development of fluorine-substituted NH2-biphenyl-diarylpyrimidines as highly potent non-nucleoside reverse transcriptase inhibitors: Boosting the safety and metabolic stability

Our recent studies for nonnucleoside reverse transcriptase inhibitors identified a highly potent compound JK-4b against WT HIV-1 (EC50 = 1.0 nmol/L), but the poor metabolic stability in human liver microsomes (t 1/2 = 14.6 min) and insufficient selectivity (SI = 2059) with high cytotoxicity (CC50 = 2.08 μmol/L) remained major issues associated with JK-4b. The present efforts were devoted to the introduction of fluorine into the biphenyl ring of JK-4b, leading to the discovery of a novel series of fluorine-substituted NH2-biphenyl-diarylpyrimidines with noticeable inhibitory activity toward WT HIV-1 strain (EC50 = 1.8-349 nmol/L). The best compound 5t in this collection (EC50 = 1.8 nmol/L, CC50 = 117 μmol/L) was 32-fold in selectivity (SI = 66,443) compared to JK-4b and showed remarkable potency toward clinically multiple mutant strains, such as L100I, K103N, E138K, and Y181C. The metabolic stability of 5t was also significantly improved (t 1/2 = 74.52 min), approximately 5-fold higher than JK-4b in human liver microsomes (t 1/2 = 14.6 min). Also, 5t possessed good stability in both human and monkey plasma. No significant in vitro inhibition effect toward CYP enzyme and hERG was observed. The single-dose acute toxicity test did not induce mice death or obvious pathological damage. These findings pave the way for further development of 5t as a drug candidate.

Pathophysiology of obesity and its associated diseases

The occurrence of obesity has increased across the whole world. Many epidemiological studies have indicated that obesity strongly contributes to the development of cancer, cardiovascular diseases, type 2 diabetes, liver diseases and other disorders, accounting for a heavy burden on the public and on health-care systems every year. Excess energy uptake induces adipocyte hypertrophy, hyperplasia and formation of visceral fat in other non-adipose tissues to evoke cardiovascular disease, liver diseases. Adipose tissue can also secrete adipokines and inflammatory cytokines to affect the local microenvironment, induce insulin resistance, hyperglycemia, and activate associated inflammatory signaling pathways. This further exacerbates the development and progression of obesity-associated diseases. Although some progress in the treatment of obesity has been achieved in preclinical and clinical studies, the progression and pathogenesis of obesity-induced diseases are complex and unclear. We still need to understand their links to better guide the treatment of obesity and associated diseases. In this review, we review the links between obesity and other diseases, with a view to improve the future management and treatment of obesity and its co-morbidities.

Normative wideband absorbance measures in children: a cross-sectional study / 中华耳鼻咽喉头颈外科杂志

Objective: This study was to investigate the main characteristics and related factors of wideband absorbance (WBA) in children with normal hearing and to obtain age-specific reference range of WBA. Methods: 384 children between 0-12 years old (615 ears) who visited the Beijing Children's Hospital, Capital Medical University from October 2019 to February 2021 were enrolled, including 230 males (376 ears) and 154 females (239 ears), with totally 306 left ears and 309 right ears. Wideband tympanometry (WBT) was performed and normative WBA data were analyzed by SPSS 24.0 statistical software. Repeated measures and multivariate analysis of variance were applied to the data from 16 points at 1/3-octave frequencies (226, 324, 408, 500, 667, 841, 1 000, 1 297, 1 682, 2 000, 2 670, 3 364, 4 000, 5 339, 6 727 and 8 000 Hz) to evaluate the effects of frequency, age, external auditory canal pressures, gender and ear on WBA. Results: According to the WBT frequency-absorbance curve, the subjects were divided into seven groups: 1-month old group, 2-month old group, 3-month old group, 4-5 month old group, 6-24 month old group,>2-6 year old group and>6-12 year old group. The WBA of normal-hearing children underwent a series of developmental changes with age at both ambient pressure and tympanometric peak pressures. WBA results for 1-month group and 2-month old group exhibited a multipeaked pattern, with the peaks occurring around 2 000 and 4 897 Hz, and a notch around 3 886 Hz. WBA results for 3-month group and 4-5 month old group exhibited a single broad-peaked pattern, with the peak occurring between 2 000-4 757 Hz. The WBA of 1-month old group to 4-5 month old group decreased gradually at low frequency (226-408 Hz) and 6 727 Hz, and increased at middle to high frequency (2 670-4 000 Hz). The WBA of 6-24 month old group were significantly lower than that of 2-month old group to 4-5 month old group at all frequencies except 3 364 and 4 000 Hz. WBA results for 6-24 month old group,>2-6 year old group and>6-12 year old group exhibited a single-peaked pattern, and the peak frequency of WBA moved to the lower frequency successively. From 6-24 month old group to>6-12 year old group, the WBA gradually increased at low to middle frequencies (667-2 670 Hz) and 8 000 Hz, and decreased at middle to high frequencies (3 364-5 339 Hz). Among the 16 frequencies of all age groups, the difference between WBA under ambient pressure and tympanometric peak pressure were -0.09-0.06, and 43.75%-81.25% frequency points had statistically significant difference, which was mainly manifested in that WBA under ambient pressure were lower than that under tympanometric peak pressure at 226-1 682 Hz. There was no significant ear effect on all of the age groups. Similarly, there was no significant gender effect except for 3-month old group and 4-5 month old group. Conclusions: The WBA of normal-hearing children measured at ambient pressure and tympanometric peak pressure varied across the frequencies with age from 1 month to 12 years old, and different frequencies followed different change patterns (increase vs. decrease) in WBA. There was also significant external auditory canal pressures effect on all of the age groups. The establishment of age-specific reference range of WBA for 0-12 years old normal-hearing children in this study would be useful for clinical practice of determining normative data regarding WBT.

Structural changes of the frontal cortex in depressed mice are associated with decreased expression of brain-derived neurotrophic factor / 南方医科大学学报

OBJECTIVE@#To investigate the changes in gray matter volume in depressive-like mice and explore the possible mechanism.@*METHODS@#Twenty-four 6-week-old C57 mice were randomized equally into control group and model group, and the mice in the model group were subjected to chronic unpredictable mild stimulation (CUMS) for 35 days. Magnetic resonance imaging was performed to examine structural changes of the grey matter volume in depressive-like mice. The expression of brain-derived neurotrophic factor (BDNF) in the grey matter of the mice was detected using Western blotting and immunofluorescence staining.@*RESULTS@#Compared with the control mice, the mice with CUMS showed significantly decreased central walking distance in the open field test (P < 0.05) and increased immobile time in forced swimming test (P < 0.05). Magnetic resonance imaging showed that the volume of the frontal cortex was significantly decreased in CUMS mice (P < 0.001, when the mass level was greater than or equal to 10 756, the FDRc was corrected with P=0.05). Western blotting showed that the expression of mature BDNF in the frontal cortex was significantly decreased in CUMS mice (P < 0.05), and its expression began to decrease after the exposure to CUMS as shown by immunofluorescence staining. The volume of different clusters obtained by voxel-based morphometry (VBM) analysis was correlated with the expression level of mature BDNF detected by Western blotting (P < 0.05).@*CONCLUSION@#The decrease of frontal cortex volume after CUMS is related with the reduction of mature BDNF expression in the frontal cortex.

Effects of low centrifugal force on sperm morphology of normal and non-liquefied semen samples / 临床检验杂志

Objective To explore the effects of low centrifugal force on sperm morphology analysis of normal and non-liquefied semen samples.Methods A total of 169 patients who underwent semen routine examination and sperm morphology analysis at the Reproduc-tive Center of the 904th Hospital of the Joint Logistics Support Force of PLA from January 2021 to December 2021 were collected as the study subjects.Among them,126 were the patients with normal semen routine parameters,and 43 with non-liquefied semen.The mor-phological analysis results of 126 normal semen samples treated by the direct smear method and centrifugation methods under two differ-ent of low centrifugal force were compared.The results of morphological analysis for the 43 non-liquefied semen samples treated by the direct smear method after promoting liquefaction and centrifugation were compared.Results The percentage of normal morphological sperm[(9.39±0.50)％]obtained by centrifugation of 340xg centrifugal force for 10 min was significantly lower than that[(11.08± 0.41)％]obtained by the direct smear method,and the abnormal rate of sperm in the middle segment[(62.05±1.44)％]obtained by centrifugation of 340xg centrifugal force was significantly higher than that[(57.17±0.82)％]obtained by the direct smear method.The percentage of normal morphological sperm[(9.41±0.57)％]obtained by centrifugation at 151×g centrifugal force was significantly lower than that obtained by the direct smear method,and the abnormal rate of sperm in the middle segment[(60.95±1.17)％]ob-tained by centrifugation at 151×g centrifugal force was significantly higher than that obtained by the direct smear method.For non-lique-fied semen samples,the percentage of normal morphological sperm obtained by centrifugation at 340xg centrifugal force for 10 min was[(9.61±0.60)％],while that obtained by the direct smear method after promoting liquefaction was[(11.10±0.73)％].The difference was not statistically significant between the two methods.The abnormal rate of sperm in the middle segment[(60.21±1.51)％]ob-tained by the centrifugation method was also not significantly different from that[(57.36±1.34)％]obtained by the direct smear meth-od after promoting liquefaction.Conclusion Both kinds of low centrifugal force affected the sperm morphology.However,the low cen-trifugal force has no significant effect on the non-liquefied semen.In the clinical application of sperm morphology analysis,the nature of male semen should be referred to determine whether the centrifugation is need or not and the magnitude of centrifugal force.

Effectiveness and security of anisodine hydrobromide tablets in treating nonarteritic anterior ischemic optic neuropathy: a Chinese multicenter nonrandomized controlled study / 中华实验眼科杂志

Objective:To evaluate the efficacy and safety of oral anisodine hydrobromide tablets in the treatment of nonarteritic anterior ischemic optic neuropathy (NAION).Methods:A multicenter nonrandomized controlled trial was conducted.A total of 282 acute NAION patients (282 eyes) were recruited from 16 hospitals in China from July 2020 to May 2021.Patients were divided into two groups according to treatment methods, which were control group (124 cases, 124 eyes) receiving regular treatment including citicoline sodium plus Ginkgo biloba leaf liquid extract or Ginkgo biloba leaf extract tablets plus mecobalamin, and experimental group (158 cases, 158 eyes) receiving treatment in control group plus oral anisodine hydrobromide tablets 1 mg, twice daily for 2 to 3 months.Best corrected visual acuity (BCVA), visual field index (VFI), peripapillary retinal nerve fiber layer (pRNFL) and radial peripapillary capillary vessel density (RPC) were assessed at 1, 2, 3, and 6 months after enrollment using the standard decimal visual acuity chart, 750i Humphery visual field analyzer, Cirrus HD-OCT 4000/Cirrus HD-OCT 5000, RTVue-XR optical coherence tomography respectively.The primary outcomes were BCVA and VFI, and the secondary outcomes were pRNFL, RPC, and the side effects during the follow-up.The study adhered to the Declaration of Helsinki.All patients were fully informed about the treatment and purpose of this study and voluntarily signed the informed consent form.The study protocol was approved by Chinese PLA General Hospital (No.S2020-021-01). Results:In all, 242 patients (242 eyes) completed the follow-up of BCVA, and 98 patients (98 eyes) completed the VFI follow-up.In terms of visual function, BCVA and VFI improved significantly over time in the two groups, and BCVA and VFI were better in experimental group than in control group at various follow-up time points (all at P<0.05). In terms of structure, pRNFL gradually decreased in both groups with the extension of treatment, and pRNFL was significanthy thinner in experimental group than in control group at various follow-up time points (all at P<0.05). There was no significant difference in RPC between the two groups at the last follow-up ( P>0.05). There were two cases with side effects and one case was discontinued due to side effects 25 days after enrollment. Conclusions:Oral anisodine hydrobromide can improve visual acuity and visual field in NAION and accelerate the regression of optic disc edema, with good safety.

Genetic inhibition of MLKL exerts protective effects against cognitive impairment and neuroinflammation in a cerebral small vessel disease model / 中国药理学与毒理学杂志

OBJECTIVE To identify the role of mixed lineage kinase domain like protein(MLKL)in cerebral small vessel disease(CSVD)and explore the underlying mechanism.METHODS Transient bilateral common carotid artery occlusion(tBCCAO)was used to establish a mouse model of CSVD.Immunofluorescence staining and Western blotting were used to observe the expres-sions of RIPK3/MLKL signaling molecules in brain tissues at 7,14 and 28 d after tBCCAO.Open field test,rotarod test,Y-maze and novel object recognition test were used to observe the effect of MLKL knockout on cognitive func-tion after tBCCAO.Blood-brain barrier(BBB)disruption was observed by sodium fluorescein permeability test and the expressions of tight junction proteins.Immunoflu-orescence staining and Western blotting were used to detect the expression of microglia marker Iba-1,astro-cyte marker GFAP,and NLRP3/Caspase-1 signaling mol-ecules in the hippocampus of CSVD mice.ELISA was used to detect the level of inflammatory factors(TNF-α,IL-1β,IL-18)in hippocampus.RESULTS The expres-sions of RIPK3/MLKL signaling molecules increased in cortex and hippocampus after tBCCAO,especially on day 14.The expression of pMLKL mainly increased in neurons,glia cells and endothelial cells in CSVD mice.MLKL knockout improved the cognitive functions such as motor learning,spatial learning and working memory,and object recognition ability in CSVD mice.MLKL knock-out alleviated the leakage of sodium fluorescein and attenuated the down-regulation of tight junction proteins at 1 d and 14 d after tBCCAO.At 14 d after tBCCAO,MLKL knock out inhibited the activations of microglia and astrocytes,attenuated the expressions of NLRP3/cas-pase-1 molecules,and decreased the levels of inflamma-tory factors in the hippocampus of mice.CONCLUSION Genetic inhibition of MLKL exerts protective effects against cognitive impairment by ameliorating BBB dam-age and neuroinflammation in a mouse cerebral small vessel disease model.

Blood flow restriction can improve knee proprioception and motor coordination after anterior cruciate ligament reconstruction / 中华物理医学与康复杂志

Objective:To explore any effect of blood flow restriction during exercise on knee proprioception and motor coordination after anterior cruciate ligament (ACL) reconstruction.Methods:Thirty patients recovering from ACL reconstruction were randomly divided into an experimental group and a control group, each of 15. Both groups were given routine rehabilitation training, while the experimental group was additionally provided with 45 minutes of training with blood flow restriction, 3 times a week for 8 weeks. The blood flow restriction training involved constant pressure in the groin while the patient performed knee flexion and extension resistance training, squats, alternate knee flexion and extension and ergometer cycling. Before and after the intervention, both groups′ knee function, proprioception and lower limb motor coordination were evaluated using the Lysholm knee scale, the Humac isokinetic measurement system and surface electromyography.Results:Before the experiment there were no significant differences between the two groups in any of the measurements. After the intervention, both groups′ average Lysholm score had improved significantly, and errors in reproducing a knee angle had decreased significantly. Significantly better improvement was observed in the observation group than in the control group. That group′s average coordinated contraction rate on the affected side in extension and flexion was also significantly better than the control group′s ave-rage. Indeed, there were no significant differences in the contraction rates between the healthy and affected sides.Conclusions:Training with restricted blood flow can significantly improve knee function, proprioception and motor coordination after anterior cruciate ligament reconstruction.

Effect of robot-assisted bilateral task-oriented upper limb training on upper limb function of patients with stroke / 中国康复理论与实践

ObjectiveTo investigate the effect of robot-assisted bilateral task-oriented upper limb training on upper limb function of patients with stroke. MethodsFrom January, 2021 to June, 2022, 52 patients with stroke in the Second Hospital of Jiaxing were enrolled and randomized into control group (n = 26) and experimental group (n = 26). Both groups received routine treatment. The control group added conventional bilateral upper limb training, while the experimental group added robot-assisted bilateral task-oriented upper limb training, for six weeks. Before and after treatment, the upper limb motor function and activities of daily living were assessed with Fugl-Meyer Assessment-Upper Extremity (FMA-UE) and modified Barthel Index (MBI). The upper limb proprioception test system was used to measure the average trace error (ATE) and test execution time of the upper limb. ResultsBefore treatment, there was no significant difference in the scores of FMA-UE and MBI, and ATE and test execution time between two groups (P > 0.05). After treatment, all the indexes improved significantly in both groups (|t| > 5.400, P < 0.001), and were better in the experimental group than in the control group (|t| > 2.123, P < 0.05). ConclusionRobot-assisted bilateral task-oriented upper limb training could facilitate to improve the upper limb motor function and activities of daily living of stroke patients.

Changes in eyebrow position following blepharoptosis surgery

Background@#The position of the eyebrows is critical when planning blepharoptosis surgery. However, insufficient scholarly attention has been paid to the details of postoperative eyebrow height changes at each anatomical landmark. This study investigated the effect of blepharoptosis surgery on brow height and evaluated the change in brow position. @*Methods@#After a retrospective review of 247 patients, this study analyzed 53 patients (106 eyelids) who underwent levator and Müller’s complex advancement between March 2010 and January 2022. Brow heights were measured from the distance between the upper brow margin of each landmark and horizontal line of pupillary center on a digital photograph. @*Results@#The mean change of eyebrow lowering was 1.54 mm (P<0.001) at the medial canthus, 1.29 mm (P<0.001) at the medial limbus, 1.44 mm (P<0.001) at the center of the pupil, 1.40 mm (P<0.001) at the lateral limbus, 1.15 mm (P=0.001) at the lateral canthus, and 0.75 mm (P=0.021) at the lateral eyebrow end. The brow change was most prominent at medial canthus and least prominent at the lateral brow end. The preoperative brow position was only statistically significant factor predicting brow height descent after surgery according to multiple linear regression analysis (R2=0.305, B=–0.375, P<0.001). @*Conclusions@#The eyebrows lowered in most patients after blepharoptosis surgery. The preoperative brow position is the most important factor in predicting the change in brow height after blepharoptosis surgery.

Investigation of the tissue-damaging effects and mechanisms of neutrophil-derived MMP-8 in Fusarium keratitis / 中华实验眼科杂志

Objective:To investigate the mechanism of tissue damage caused by neutrophil matrix metalloproteinase-8 (MMP-8) in Fusarium keratitis. Methods:A total of 108 male C57BL/6J SPF grade mice, 6-8 weeks old, were selected to establish a model of Fusarium keratitis (FK) in the right eyes.Corneal inflammation in mice was observed and scored under a slit lamp microscope.Based on the corneal inflammation scores, the modeling eyes were divided into 0, 12, 24, 48, and 72-hour groups post-modeling.At the corresponding time points, mice were euthanized, and corneal tissues were collected.The expressions of MMP-8, adenylate-activated protein kinase (AMPKα) and its serine 172-site phosphorylated form (p-AMPKα) proteins in corneal tissues were detected by Western blot.The neutrophil count in mice corneal tissues at each time point was determined using hematoxylin and eosin staining.The co-localization of neutrophils and MMP-8 protein in the cornea was observed by immunofluorescence staining.In the in vitro corneal collagen degradation experiment, corneal tissues were divided into MMP-8 group, buffer group, and normal saline group, which were treated with 100 μl of activated recombinant MMP-8, detection buffer, and normal saline, respectively.Hydroxyproline content in corneal tissues was determined using a hydroxyproline assay kit, and the mass fractions of hydroxyproline were compared among the groups.Peripheral blood neutrophils were isolated from human blood samples, and Fusarium spores were collected for experiments.Human neutrophils were divided into four groups, negative control group (cultured neutrophils), co-culture group (neutrophils co-cultured with spores), AICAR-treated group (neutrophils co-cultured with spores and treated with p-AMPK protein kinase activator AICAR), and compound C-treated group (neutrophils co-cultured with spores and treated with the inhibitor compound C).The MMP-8 protein expression levels in each group of human neutrophils were assessed via immunofluorescence staining.The use and care of animals complied with the ARVO statement and Regulations for the Administration of Affairs Concerning Experimental Animals.The animal experiment protocol was approved by the Animal Ethics Committee of Henan Eye Hospital (No.HNEECA-2017-04-02).One healthy adult volunteer was selected and 10 ml of peripheral venous blood was collected.The clinical study protocol was approved by the Clinical Ethics Committee of Henan Eye Hospital (No.HNEECKY-2019[16]). Results:At 24 hours post-modeling, corneal opacification was observed in the modeling eyes, and corneal perforation occurred in 72-hour post-modeling group.The corneal inflammation scores in 24, 48, and 72-hour post-modeling groups were all higher than those in 12-hour post-modeling group, and the differences were statistically significant (all at P<0.001).The relative expression levels of MMP-8 protein in the cornea were higher in 12, 24, and 48-hour post-modeling groups compared to 0-hour group, with statistically significant differences (all at P<0.001).There was a moderate positive correlation between the relative expression level of MMP-8 protein in the cornea and the inflammation scores of the modeling eye ( rs=0.50, P<0.05).In the cornea, the p-AMPKα (Thr 172)/AMPKα ratio was higher in 24, 48, and 72-hour post-modeling groups than in 0-hour group, and the differences were statistically significant (all at P<0.05).The p-AMPKα(Thr 172)/AMPKα ratio in the cornea was moderately positively correlated with the relative expression level of MMP-8 protein ( r=0.54, P<0.01).The number of neutrophils in the cornea was significantly higher in 24, 48, and 72-hour post-modeling groups than in 0-hour group, with statistically significant differences (all at P<0.001).The number of neutrophils in the cornea was strongly positively correlated with the inflammation score ( rs=0.77, P<0.001), and was moderately positively correlated with the relative expression level of MMP-8 protein ( r=0.56, P<0.05).MMP-8 protein expression in the cornea of the modeling eyes showed a high degree of co-localization with neutrophils.The hydroxyproline content in the cornea was (0.52±0.02)μg/mg, (0.51±0.03)μg/mg, and (0.27±0.02)μg/mg in buffer group, normal saline group and MMP-8 group, respectively, with a significant overall difference among them ( F=156.63, P<0.01).The corneal hydroxyproline content was lower in MMP-8 group compared to buffer and normal saline groups, and the differences were statistically significant (all at P<0.05).In the experiment involving the infection of cultured Fusarium spores with human neutrophils, the fluorescence intensity of MMP-8 expression was significantly higher in AICAR-treated group than in negative control group and compound C-treated group, with statistically significant differences (all at P<0.05). Conclusions:The MMP-8 secreted by neutrophils in mice with fungal keratitis can degrade corneal stromal collagen fibers, leading to corneal opacification or perforation.The variations in MMP-8 protein expression levels in human neutrophils may be associated with AMPK activation.

Preliminary exploration of SpyGlass direct visualization system in diagnosis and treatment of biliary stricture after liver transplantation / 器官移植

Objective To preliminarily evaluate the application value of SpyGlass direct visualization system in the diagnosis and treatment of biliary stricture after liver transplantation. Methods Clinical data of 4 patients presenting with biliary stricture after liver transplantation who underwent SpyGlass direct visualization system examination were collected. The examination, treatment and prognosis of biliary stricture were analyzed. Results The examination results of color Doppler ultrasound, magnetic resonance cholangiopancreatography (MRCP) and endoscopic retrograde cholangiopancreatography (ERCP) in 4 patients suggested biliary anastomotic stricture with intrahepatic biliary dilatation, and 2 of them were complicated with intrahepatic biliary calculi. Repeated placement of biliary stent under ERCP yielded poor effect in 3 cases. SpyGlass direct visualization system examination hinted biliary anastomotic stricture in 4 patients, 3 cases of intrahepatic biliary dilatation, 3 cases of intrahepatic biliary calculi, 2 cases of purulent bile and 3 cases of floccules within the biliary tract, 1 case of congestion and edema of biliary tract wall and 2 cases of local epithelial necrosis and stiffness changes of intrahepatic biliary tract wall. The wire could not be inserted in 1 patient due to severe biliary anastomotic stricture. Four patients were treated with biliary stricture resection + biliary stone removal + biliary end-to-end anastomosis, biliary stricture resection + biliary-intestinal anastomosis, ERCP lithotomy + biliary metal stent implantation, and biliary metal stent implantation + percutaneous transhepatic bile duct lithotomy, respectively. Relevant symptoms were relieved without evident complications. All patients survived during the follow-up until the submission date. Conclusions Compared with traditional imaging examination, SpyGlass direct visualization system may more directly display the morphological characteristics of biliary tract wall and structural changes within biliary tract cavity, which is an effective examination tool for biliary stricture after liver transplantation. In addition, individualized treatment methods may be adopted for different biliary tract diseases, which is expected to improve clinical prognosis of patients.