ObjectiveBased on the concept of quality by design（QbD）， the processing process of calcined Pyritum was optimized， and its X-ray diffraction（XRD） fingerprint was established. MethodsThe safety， effectiveness and quality controllability of calcined Pyritum were taken as the quality profile（QTPP）， the color， hardness， metallic luster， phase composition， the contents of heavy metals and hazardous elements were taken as the critical quality attributes（CQAs）， and the calcination temperature， calcination time， paving thickness and particle size were determined as the critical process parameters（CPPs）. Differential thermal analysis， X-ray diffraction（XRD） and inductively coupled plasma mass spectrometry（ICP-MS） were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then， based on the criteria importance through intercriteria correlation（CRITIC）-entropy weight method， the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process， and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures， and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS₂ to Fe₇S₈ during the process of temperature increase， the color gradually deepened from dark yellow， and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows：calcination temperature at 750 ℃， calcination time of 2.5 h， paving thickness of 3 cm， particle size of 0.8-1.2 cm， vinegar quenching 1 time［Pyritum-vinegar（10∶3）］. After calcination， the internal structure of Pyritum was honeycomb-shaped， which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96， and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing， the dissolution of the effective components in Pyritum increases， and the contents of heavy metals and harmful substances decrease， reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible， and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.

ObjectiveBased on the concept of quality by design（QbD）， the processing process of calcined Pyritum was optimized， and its X-ray diffraction（XRD） fingerprint was established. MethodsThe safety， effectiveness and quality controllability of calcined Pyritum were taken as the quality profile（QTPP）， the color， hardness， metallic luster， phase composition， the contents of heavy metals and hazardous elements were taken as the critical quality attributes（CQAs）， and the calcination temperature， calcination time， paving thickness and particle size were determined as the critical process parameters（CPPs）. Differential thermal analysis， X-ray diffraction（XRD） and inductively coupled plasma mass spectrometry（ICP-MS） were used to analyze the correlation between the calcination temperature and CQAs of calcined Pyritum. Then， based on the criteria importance through intercriteria correlation（CRITIC）-entropy weight method， the optimal processing process of calcined Pyritum was optimized by orthogonal test. Powder XRD was used to analyze the phase of calcined Pyritum samples processed according to the best process， and the mean and median maps of calcined Pyritum were established by the superposition of geometric topological figures， and similarity evaluation and cluster analysis were carried out. ResultsThe results of single factor experiments showed that the physical phase of Pyritum changed from FeS₂ to Fe₇S₈ during the process of temperature increase， the color gradually deepened from dark yellow， and the contents of heavy metals and harmful elements decreased. The optimized processing process of calcined Pyritum was as follows：calcination temperature at 750 ℃， calcination time of 2.5 h， paving thickness of 3 cm， particle size of 0.8-1.2 cm， vinegar quenching 1 time［Pyritum-vinegar（10∶3）］. After calcination， the internal structure of Pyritum was honeycomb-shaped， which was conducive to the dissolution of active ingredients. XRD fingerprints of 13 batches of calcined Pyritum characterized by 10 common peaks were established. The similarities of the relative peak intensities of the XRD fingerprints of the analyzed samples were>0.96， and it could effectively distinguish the raw products and unqualified products. ConclusionTemperature is the main factor affecting the quality of calcined Pyritum. After processing， the dissolution of the effective components in Pyritum increases， and the contents of heavy metals and harmful substances decrease， reflecting the function of processing to increase efficiency and reduce toxicity. The optimized processing process is stable and feasible， and the established XRD fingerprint can be used as one of the quality control standards of calcined Pyritum.

Objective:To develop a recombinant protein vaccine based on KPC-2, a drug resistance target in Klebsiella pneumoniae, and evaluate its immunogenicity, protective efficacy and mechanism in a mouse model of pneumonia. Methods:KPC-2 was expressed in Escherichia coli and purified using GST affinity chromatography. A recombinant protein vaccine was prepared with KPC-2 and used to immunize New Zealand rabbits through subcutaneous injection. Serum samples were isolated from cardiac blood and Protein G chromatography was used to purify polyclonal antibodies against KPC-2. Opsonophagocytic killing assay was used to assess the bactericidal activity of the polyclonal antibodies in vitro. Female BALB/c mice were immunized three times with the recombinant protein vaccine, and the titers of specific IgG antibodies in serum were measured by indirect ELISA. One week after the last vaccination, the mice were infected with Klebsiella pneumoniae strain SRT through tracheal intubation, and received a single intravenous dose of meropenem (0.1 mg) 1 h later. The protective efficacy of the KPC-2 recombinant protein vaccine was evaluated by comparing the survival rates, bacterial colonization and histopathological changes between vaccine group and adjuvant group as well as the survival rates between meropenem group and normal saline group. Moreover, the protective efficacy of polyclonal antibodies against KPC-2 was evaluated through passive immunization. Results:The level of specific IgG antibodies in serum was significantly higher in the vaccine group than in the adjuvant group ( t=4.325, P<0.05). The survival rate in the vaccine group was also higher than that of the adjuvant group [70% (7/10) vs 10% (1/10), P<0.05]. Furthermore, lung inflammation was less severe and bacterial burden was reduced in the vaccine group as compared with those of the control group ( t=3.127, P<0.05). Both active and passive vaccination strategies demonstrated strong protective efficacy against Klebsiella pneumoniae infection, and had a synergistic effect when used in combination with antibiotic therapy. The polyclonal antibodies against KPC-2 had bactericidal activity in vitro ( t=5.427, P<0.05). Conclusions:The prepared KPC-2 vaccine has better immunogenicity and protective efficacy. It can induce strong humoral immune responses. This study suggest that drug resistance target may be used as a candidate antigen for future vaccine development.

Objective:To explore the current situation of nursing human caring in hospital wards and analyze its influencing factors, so as to facilitate the development of nursing human caring practice.Methods:From July to November 2022, a total of 107 hospitals were surveyed through stratified convenience sampling method, and 4 072 ward nursing managers were recruited to finish the general information questionnaire and the ward nursing human caring status questionnaire. The general information included the region, class and type of the hospital, etc. The ward nursing human caring status questionnaire included 38 items in 5 dimensions of nursing human caring system and process, humanistic quality and training of nursing staff, humanistic environment and facilities, human caring procedures and measures, and human caring quality evaluation and improvement, with a full score of 190 points. Descriptive statistics were used to analyze the general data, independent samples t-test, ANOVA and correlation analysis were used to analyze the factors influencing the current status of nursing human caring in the ward, while multiple linear regression analysis was used to conduct a multivariate analysis. Results:The score of nursing human caring in hospital wards was 156.91±27.78. Whether the hospital had carried out nursing human caring pilot(demonstration) wards, whether the ward had previously been a hospital nursing human caring pilot(demonstration) nursing unit, the type of ward, and whether nursing managers had participated in human caring training were the influencing factors of the implication of nursing humanistic caring in wards( P<0.05). Conclusions:The practice of nursing human caring in hospital wards is at a good level, but needs to be further strengthened. Nursing managers should take systematically strategies to promote the development of nursing human caring practice.

Humans ; Child ; Retrospective Studies ; Thrombocytosis/etiology*

Humans ; Granuloma, Pyogenic/pathology* ; Hemangioma, Capillary

The erythropoietin-producing hepatocellular receptor (Eph receptor) family is the largest subfamily in the receptor tyrosine kinase (RTK) families which mediates cell morphology, adhesion, movement, proliferation, survival, and differentiation by its bidirectional signals coupled with Ephrin ligands. EphA2 receptor is an important isoform which is involved in the pathological changes in cataract, breast cancer, etc. Previous studies found that the kinase domain of the EphA2 receptor binds to the plasma membrane, and its kinase activity is regulated by the plasma membrane. However, it is still unclear that the impact of the adjacent SAM domain on the membrane binding and kinase activities of kinase domain. In this study we purified the cytoplasmic kinase-SAM tandem of the EphA2 receptor by co-expression with the phosphatase PTP1B 1-301 fragment. Our results showed that the SAM domain of EphA2 receptor can further enhance the interaction between the kinase domain and liposomes (4 mg/mL) by 6 folds (P<0. 001). And the phosphorylation of kinase-SAM tandem can enhance its lipid (4 mg/mL) binding ability by 2. 5 folds (P < 0. 05). In addition, the lipid binding ability and tyrosine phosphorylation activities of kinase domain are mutual promoted, which creating a positive feedback loop in the two biological processes. In conclusion, our studies indicate that the kinase domain and the adjacent SAM domain can function as an intact unit, whose lipid binding ability and kinase activity are quite different from the individual kinase domain. Therefore, our results provide a biochemical basis for better understanding of the regulation mechanism of other Eph receptors in its kinase domain.

Objective:To investigate the causes of secondary glaucoma after vitrectomy for familial vitreous amyloidosis associated with transthyretin (TTR) gene Gly83Arg mutation.Methods:A retrospective case study. From January 2008 to January 2020, 13 cases (23 eyes) with hereditary vitreous amyloidosis and treated by vitrectomy in the Affiliated Hospital of Zunyi Medical University were collected. Among them, there were 7 males with 12 eyes and 6 females with 11 eyes. The average age was 43.0±4.8 years. All the affected eyes underwent standard three-channel vitrectomy through the flat part of the ciliary body. According to whether complete vitreous detachment (PVD) was formed during the operation, it was divided into complete PVD group and incomplete PVD group; according to the occurrence time of secondary glaucoma and vitreous amyloidosis after surgery, it was divided into 1-12 months group and 13-36 months group, >37 months group. The average follow-up time after surgery was 36.7±6.0 months. The incidence of secondary glaucoma and the recurrence rate of vitreous amyloidosis between groups were compared by χ2 test; the correlation between recurrence of vitreous amyloidosis and secondary glaucoma after surgery was analyzed by Spearman rank correlation analysis. Results:Among the 23 eyes, there were 8 eyes in the complete PVD group and 15 eyes in the incomplete PVD group, respectively. Vitreous amyloidosis recurred in 15 eyes (65.22%, 15/23) after surgery. There were 14 (93.30%, 14/15) and 1 (6.70%, 1/15) eyes in the incomplete PVD group and the complete PVD group, respectively; the comparison of the recurrence rate of vitreous amyloidosis between the two groups was statistically significant ( χ2=11.676, P<0.01). 1-12 months group, 13-36 months group, >37 months group included 1 (4.35%, 1/23), 12 (52.17%, 12/23), 2 (8.70%, 2/23) Only eye. The recurrence rate in the 13-36 months group was significantly higher than that in the 1-12 months group and >37 month group. Secondary glaucoma occurred in 11 eyes (47.80%, 11/23) after surgery. 1-12 months group, 13-36 months group, above 37 months group were 1 (4.35%, 1/23), 8 (34.78%, 8/23), 2 (8.70%, 2/23) eyes. The incidence of secondary glaucoma in the 13-36 months group was higher than that in the 1-12 months group and >37 months group. Among 11 eyes with secondary glaucoma, 10 eyes had recurrence of vitreous amyloidosis after surgery, and 1 eye had no recurrence. The results of Spearman rank correlation analysis showed that there was a positive correlation between the recurrence of vitreous amyloidosis and the occurrence of secondary glaucoma ( rs=0.516, P=0.012). Conclusion:The incidence of secondary glaucoma after vitrectomy in a family with vitreous amyloidosis caused by the Gly83Arg mutation of TTR gene is higher, and its occurrence is significantly positively correlated with the recurrence of vitreous amyloidosis.

Objective:To investigate the clinical features of Herpes simplex virus encephalitis(HSE) with cerebral hematoma as the prominent manifestation and the significanc of metagenomic next-generation sequencing (mNGS) in the diagnosis of HSE.Methods:The clinical manifestations, diagnostic process, clinical treatment and prognosis of a case of HSE with cerebral hematoma as the prominent manifestation at Shanghai Children′s Medical Center Affiliated to Shanghai Jiaotong University School of Medicine in June 2019 were retrospectively analyzed.The relevant literatures were also searched and reviewed.Results:A 4-year-old boy presented with slight fever, headache, convulsion and vomiting was considered to have intracranial space-occupying lesions and possible intratumoral hemorrhage after undergoing imaging examination at a local hospital.The patient was checked by head CT in Shanghai Children′s Medical Center Affiliated to Shanghai Jiaotong University School of Medicine, which showed that there were many bleeding foci in the brain, indicating the possibility of complications of blood system diseases.Therefor the child was given the examination of blood routine and coagulation routine, but the results were normal, the bone marrow cytology was negative, the cerebrospinal fluid(CSF) of lumbar puncture was biochemically normal, and mNGS were 8×10 6/L.Besides, CSF smear, culture and next-generation sequencing were negative, the autoimmune encephalitis CSF testing was negative, and brain biopsy suggested inflammation.The mNGS brain tissue showed herpes simplex virus 1 was positive in two specimens, confirming the diagnosis of HSE.After 3 weeks of antiviral treatment with Aciclovir, the child′s condition improved.After a 5-month follow-up, the patient had quadriplegia and only had activities such as blinking and swallowing. Conclusions:When the intracerebral hemorrhage such as hematoma caused by encephalitis clinically can not be ruled out, the possibility of HSE should be considered, and mNGS is helpful for identifying the central ner-vous system pathogen.

Objective:To study the effect of melatonin (MEL) on the pyroptosis of hippocampus in neonatal rats with hypoxic-ischemic brain damage (HIBD), and the related mechanism.Methods:The animal model of HIBD was established by the modified Rice method.According to the random number table, a total of 105 Sprague-Dawley (SD) rats aged 7 days were divided into 7 groups (15 rats in each group): sham operation (Sham) group, model (HIBD) group, MEL treatment group (5, 10 and 20 mg/kg), phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway inhibitor (LY294002) treatment group and MEL+ LY294002 group.The hippocampus neuronal morphology and the changes of nissl bodies were observed through HE staining and nissl staining.The mRNA expression levels of Nod-like receptor family 3 (NLRP3), apoptosis-associated speck-like protein containing a card (ASC), Caspase-1, gasdermin D (GSDMD), interleukin-1β (IL-1β) and interleukin-18 (IL-18) in the left hippocampus of rats were detected by real-time fluorescence quantitative PCR.The protein expression level of the above indexes and the level of phosphorylated Akt (p-Akt) were measured by Western blot.Results:Compared with the Sham group, the number of cell layers in hippocampal CA1 region in the HIBD group decreased, the cell arrangement was irregular, and there were less nissl bodies.Besides, the mRNA expression levels of NLRP3 (1.98±0.08 vs.0.86±0.13), ASC (1.40±0.12 vs.0.81±0.07), Caspase-1 (1.46±0.10 vs.0.75±0.09), GSDMD (1.35±0.10 vs.0.81±0.10), IL-18 (1.23±0.08 vs.0.23±0.04), IL-1β (1.83±0.09 vs.0.57±0.08) and p-Akt (1.12±0.12 vs.0.54±0.07) in the HIBD group were significant higher than those in the Sham group (all P<0.05). Compared with the HIBD group, there were more cell layers in hippocampal CA1 region of the MEL group (10 mg/kg), the arrangement of cells was more regular, and the number of nissl bodies increased.The mRNA expression levels of NLRP3 (1.04±0.10), ASC (0.91±0.06), Caspase-1 (0.63±0.06), GSDMD (1.01±0.09), IL-18 (0.65±0.05) and IL-1β (0.63±0.10) in the MEL group were statistically significantly lower than those in the HIBD group (all P<0.05). Compared with the MEL group (10 mg/kg), the arrangement of cells in hippocampal CA1 region of the MEL+ LY294002 group was relatively disordered, the nissl bodies declined, the p-Akt protein level (0.87±0.09 vs.1.99±0.27) decreased significantly, and the Caspase-1(p20) protein level (0.85±0.09 vs.0.58±0.09) increased significantly (all P<0.05). Conclusions:MEL may inhibit the hippocampal pyroptosis in neonatal rats with HIBD by activating the Akt signaling pathway, thereby protecting the brain.