ObjectiveTo compare the contents and relative molecular weight distributions of proteins and polypeptides in Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules of Hirudo single medicinal preparations， to evaluate the in vitro anticoagulant， antiplatelet and fibrinolytic activities of the three preparations， and to investigate the effects of temperature， pH and digestive enzymes on the anticoagulant activities of the three preparations. MethodsThe contents of soluble proteins and polypeptides in the three preparations were determined by bicinchoninic acid assay（BCA） and Bradford method， and the relative molecular weight distributions of the three preparations were determined by electrophoresis combined with gel chromatography. The antithrombin activity of the three preparations was evaluated by fibrinogen-thrombin time（Fibg-TT） method， and their anticoagulant activities were further assessed by the elongations of activated partial thromboplastin time（APTT）， prothrombin time（PT） and thrombin time（TT）. The antiplatelet aggregation activities of the three preparations were measured by turbidimetry and the fibrinolytic activities were measured by fibrin plate method. Relative TT was used as index to investigate the effects of temperature， pH and digestive enzyme buffer on anticoagulant activities of the three preparations. ResultsAt the lowest single dosage， the contents of proteins and polypeptides were in the order of Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. Both Huoxue Tongmai capsules and Maixuekang capsules had 11 electrophoretic bands between 4.0 kDa and 90 kDa， the bands of Maixuekang capsules were more clear in the range of >25 kDa， and there was 1 obvious band at 14 kDa for the two capsules. Huoxue Tongmai capsules had one specific band at 9.0 kDa and Maixuekang capsules had one specific band at 48.0 kDa. Naoxuekang dropping pills only had 2 electrophoretic bands at 6.5 kDa and 8.5 kDa， primarily containing peptides below 2 kDa， most of which were oligopeptides. The anticoagulant activity concentrations of the three preparations exhibited a certain dose-dependent effect. At the lowest single dosage， The anticoagulant activity concentrations were ranked as Naoxuekang dropping pills>Huoxue Tongmai capsules>Maixuekang capsules. The prolongation effect of the three preparations on coagulation time was dose-dependent. At the same concentration， the prolongation effect of Naoxuekang dropping pills and Huoxue Tongmai capsules was APTT prolongation rate>TT prolongation rate>PT prolongation rate， whereas for Maixuekang capsules， the sequence was TT prolongation rate>APTT prolongation rate>PT lengthening rate. At the single minimum dosage， the order of APTT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules≈Naoxuekang dropping pills， the order of PT prolongation rate was Naoxuekang dropping pills≈Maixuekang capsules>Huoxue Tongmai capsules， and the order of TT prolongation rate was Maixuekang capsules>Huoxue Tongmai capsules>Naoxuekang dropping pills. The three preparations showed dose-dependent effects on platelet aggregation induced by adenosine diphosphate（ADP） and arachidonic acid（AA）， and the effect induced by ADP was stronger than that induced by AA. The anti-platelet aggregation effect of Naoxuekang dropping pills was significantly stronger than that of Maixuekang capsules（P<0.01）， whereas Huoxue Tongmai capsules had the effect of promoting platelet aggregation. None of the three preparations had the ability to dissolve fibrin. The anticoagulant activity of Naoxuekang dropping pills was least affected by heating， while the activities of the two capsules decreased significantly within 5 min above 80 ℃， and continued to decrease within 2 h. Compared with pure water， the anticoagulant activities of the three preparations could be increased by 1-3 times under strong acidity（pH 1-3）. In the pepsin buffer， the anticoagulant activity of Naoxuekang dropping pills could be increased by 1-3 times， while the anticoagulant activities of Huoxue Tongmai capsules and Maxuekang capsules were significantly decreased， the lowest levels were about 60% and 20%， respectively. In trypsin buffer， the anticoagulant activities of Naoxuekang dropping pills， Huoxue Tongmai capsules and Maixuekang capsules decreased significantly， and the lowest levels decreased to about 41%， 41% and 35%， respectively. ConclusionThe contents of proteins and polypeptides and relative molecular weights of the preparations derived from lyophilized fresh Hirudo powder， dried Hirudo powder and reflux extract of Hirudo decrease sequentially， and the anticoagulant activity decrease gradually， but the anticoagulant pathway is different. And the anti-platelet aggregation activity of the reflux extract is significantly enhanced. The heat resistance and gastrointestinal stability of the three preparations increase successively， and the first two are suitable for enteric-soluble preparations， while the latter is suitable for routine oral administration. The above results can provide data reference for the rationality of different preparation methods， active substances， pharmacodynamics and mechanism of Hirudo preparations.

Objective To investigate the effect of protocatechuic acid(PCA)on Toll-like receptor 4(TLR4)/nu-clear factor-kappa B(NF-κB)signal pathway in rats with ulcerative colitis(UC).Methods After modeling,rats were divided into control group,model group(5%trinitrobenzene sulfonic acid(TNBS)induction),low(L-PCA),middle(M-PCA),high(H-PCA)dose protocatechuic acid group(rats were given with PCA at doses of 20,40 and 80 mg/kg respectively),and intervention model group(LPS,rats were given with PCA at doses of 80 mg/kg and 0.8 mg/kg LPS solution intraperitoneally),and 12 rats in each group were given the drug continu-ously for 14 days.After treatment,rats were evaluated by disease activity index(DAI),and the morphology and apoptosis of colon were observed by hematoxylin eosin(HE)staining and TUNEL staining.The protein levels of Occludin,Zona occludens 1(ZO-1),claudin-1,Tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,IL-10,TLR4 and p-p65/p65 in colon were detected by Western blotting.The expression of TLR4 protein in colon was de-tected by immunohistochemical staining.Results Compared with control group,the colon of model group rat showed obvious injury,DAI score and positive rate of TUNEL were increased(P＜0.05),the relative protein ex-pression of TNF-α,IL-1β,TLR4 and p-p65 were increased(P＜0.05),the relative protein expression of Occlu-din,ZO-1,claudin-1 and IL-10 were decreased(P＜0.05).Compared with model group,colonic injury was allevi-ated in L-PCA,M-PCA and H-PCA groups,DAI score and positive rate of TUNEL were decreased(P＜0.05),the relative protein expression of TNF-α,IL-1β,TLR4 and p-p65 were decreased(P＜0.05),the relative protein expression of Occludin,ZO-1,claudin-1 and IL-10 were increased(P＜0.05).Compared with H-PCA group,co-lon injury was aggravated in LPS group,DAI score and positive rate of TUNEL were increased(P＜0.05),the rel-ative protein expression of TNF-α,IL-1β,TLR4 and p-p65 was increased(P＜0.05),the relative protein expres-sion of Occludin,ZO-1,claudin-1 and IL-10 was decreased(P＜0.05).Conclusions Protocatechuic acid can al-leviate TNBS-induced colon injury in UC rats,improve intestinal barrier function,and inhibit colon inflammatory response,the mechanism of which is related to the activation of TLR4/NF-κB signaling pathway.

Objective To analyze the characteristics of hepatitis B infection time in Baqiao Distract of Xian City, predict the incidence trend of hepatitis B in the next years with ARIMA model, and provide theoretical guidance for the adjustment of hepatitis B prevention and control strategies. Methods A total of 7173 cases of hepatitis B in our hospital from 2018 to 2023 were collected as subjects,the annual percentage of change (APC) was used to analyze the characteristics of time of hepatitis B infection in the next years, and the ARIMA optimal model was used to predict the incidence rate in 2024. Results The gross incidence of hepatitis B in Baqiao Distract of Xian City from 2018 to 2023 was 51.30 per 100 000, and the standardized incidence rate was 42.11 per 100 000. The six-year incidence rate showed an upward trend (APC=8.71%,95% CI：3.29% -9.61% , P<0.05). Chi square results showed that the standardized incidence rate of hepatitis B in men is 1.51 times than women, and the incidence rate of hepatitis B in the 30-45 age group was the highest. The prediction results showed that the number of hepatitis B cases in 2024 was 1590, which was 1.91% lower than that in 2023 (1621 cases). Conclusion The infection of hepatitis B is on the rise in Baqiao Distract of Xian City, ARIMA can be used to predict this kind of infectious disease and provide scientific guidance for the prevention of this disease.

Objective To explore the effect of cannabinoid receptor 2(CB2)on orthodontic tooth movement(OTM)rate and periodontal tissue reconstruction of pressure area in mice.Methods Thirty CB2-/-male mice and thirty littermate control WT male mice were individually accepted the orthodontic appliance at their age of 6 weeks.The mice were respectively scarified at 3 days,7 days,14 days and 21 days after the operation.Then the tooth movement distance was examined through the stereomicroscope.Hematoxylin-eosin staining was performed to explore the biological responses of periodontium at the distal mesial root pressure area.Anti-tartrate acid phospha-tase staining was performed to calculate the number and distribution of osteoclasts at the distal mesial root pressure area,and MMP-9 was evaluated by immunohistochemistry to examine the number of MMP-9(+)monocytes and multinucleated cells in the same district as the TRAP staining.Results Compared with those WT mice at 3,7,14 and 21 days,OTM distance showed a gradual increased tendency according with experimental time over 21 days.The widths of periodontal ligament on the pressure side were markedly greater in CB2-/-mice than WT mice at 7,14 and 21 days(P＜0.000 1).The numbers of TRAP positive osteoclasts were significantly greater in CB2-/-mice than those in WT mice at 14 days of OTM(P＜0.001).MMP-9 immunohistochemical staining showed that the number of MMP-9(+)monocytes and multinucleated cells was more in CB2-/-mice than that in WT mice at 14 days of OTM(P＜0.05).Conclusion The absence of CB2 accelerates orthodontic tooth movement under or-thodontic force.The absence of CB2 reinforces bone resorption in orthodontic tooth movement compressive area dur-ing orthodontic tooth movement.

Objective:To observe any effect of family rehabilitation interventions based on digital health management on elderly type 2 diabetes mellitus (T2DM) patients with sarcopenia.Methods:One hundred elderly T2DM patients with sarcopenia who had been discharged from hospital after treatment were divided into an observation group and a control group, each of 50. Both groups continued the diet control and training begun during their hospitalization, but the observation group was additionally provided with family rehabilitation based on digital health management. Before and after 3 months, the glucose and lipid metabolism and sarcopenia of both groups were evaluated with related symptom indexes, and their levels of diabetes self-management were compared.Results:Significant improvement was observed in both groups, but the average glucose and lipid metabolism indexes and sarcopenia-related symptom indexes of the observation group were significantly better than the control group′s averages. Their diabetes self-management was also significantly superior.Conclusion:Family rehabilitation based on digital health management can significantly improve glucose and lipid metabolism and muscle mass in elderly T2DM patients with sarcopenia. Such intervention is worthy of promotion and application in clinical practice.

Clinicians need to focus on various points in the diagnosis and treatment of insomnia.This article prescribed the treatment protocol based on the unique features,such as insomnia in the elderly,women experiencing specific physiologi-cal periods,children insomnia,insomnia in sleep-breathing disorder patients,insomnia in patients with chronic liver and kidney dysfunction.It pro-vides some reference for clinicians while they make decision on diagnosis,differentiation and treat-ment methods.

Objective To investigate the changes of peripheral CD100 in patients with hepatocellular carcinoma (HCC), and to assess the regulatory function of CD100 to T lymphocytes in HCC patients. Methods A prospective study was conducted. Fifty-seven HCC patients and twenty-two controls who were hospitalized in our hospital between April 2020 and July 2021 were enrolled. Anti-coagulant peripheral blood was collected. Plasma and peripheral blood mononuclear cells (PBMC) were isolated. Plasma soluble CD100 (sCD100) level was measured by enzyme-linked immunosorbent assay. Membrane-bound CD100 (mCD100) expression on CD4 + and CD8 + T lymphocytes was measured by flow cytometry. PBMC from HCC patients were stimulated with recombinant human CD100. Cellular proliferation was measured by cell counting kit-8. Different types of T helper cells (Th cells) and cytotoxic T cells (Tc cells) were assessed by flow cytometry. Perforin and granzyme B secretion by alpha fetoprotein (AFP) specific CD8 + T lymphocytes was assessed by enzyme-linked immunospot assay. CD8 + T lymphocytes were purified from HCC patients, and were stimulated with recombinant human CD100. Stimulated CD8 + T lymphocytes were co-cultured with HepG2 cells. AFP specific CD8 + T lymphocytes-induced HepG2 cell death was investigated. Student's t test or paired t test was used for comparison of normally distributed continuous data between two groups. Mann-Whitney U test was used for comparison of abnormally distributed continuous data between two groups. Chi square test was used for comparison of categorial data between two groups. Results Plasma sCD100 level was lower in HCC group when compared with control group ((2.73±0.58)ng/mL vs(3.33±0.84)ng/mL, t =3.584, P < 0.001). HCC group had higher percentage of mCD100 + cells (55.57%±11.33% vs 43.67%±6.40%, t =4.636, P < 0.001) and elevated mCD100 mean fluorescent intensity (MFI) (294.80±74.01 vs 255.00±74.01, t =2.126, P =0.037) within CD4 + T lymphocytes when compared with control group. Similarly, HCC group also had higher percentage of mCD100+ cells (48.65%±7.71% vs 41.74%±4.77%, t =3.914, P < 0.001) and elevated mCD100 MFI (289.20±89.30 vs 246.10±60.73, t =2.082, P =0.041) within CD8 + T lymphocytes when compared with control group. There were no significant differences of either cellular proliferation or T lymphocytes percentage between PBMC with and without recombinant human CD100 stimulation in HCC patients (all P > 0.05). The percentages of CD4 + IFNγ + Th1 cells, CD4 + IL-17A + Th17 cells, CD4 + IL-22 + Th22 cells, and CD8 + IFNγ + Tc1 cells were notably increased in response to CD100 stimulation when compared with no CD100 stimulation ( t =2.608、5.663、4.113、4605, all P < 0.05). There were no remarkably differences of either CD8 + IL-17A + Tc17 or CD8 + IL-22 + Tc22 cell frequency between PBMCs with and without recombinant human CD100 stimulation in HCC patients (all P > 0.05). Perforin and granzyme B secretion by AFP specific CD8 + T lymphocytes were significantly elevated in response to CD100 stimulation when compared with no CD100 stimulation in HLA-A02 restricted HCC patients ( P < 0.05). AFP specific CD8 + T lymphocytes-induced HepG2 cell death was also increased in response to CD100 stimulation ( t =6.794、2.308, both P < 0.05). Conclusion There was an imbalance between sCD100 and mCD100 on T lymphocytes in HCC patients. Reduced sCD100 level might be insufficient for maintenance of T lymphocytes activity, leading to the immunotolerance in HCC.

OBJECTIVE: To explore the clinical characteristics and genetic basis for a child with X-linked lissencephaly with abnormal genitalia (XLAG). METHODS: A child with XLAG who had presented at the Third Affiliated Hospital of Zhengzhou University in May 2021 was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to high-throughput sequencing. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the result was analyzed by using bioinformatic software. RESULTS: The child was found to have harbored a hemizygous c.945_948del variant in exon 2 of the ARX gene, which as a frameshifting variant has resulted in a truncated protein. His mother was found to be heterozygous for the variant, whilst his father was of wild type. The variant was unreported previously. CONCLUSION The hemizygous c.945_948del variant of the ARX gene probably underlay the XLAG in this patient. Above finding has provided a basis for the diagnosis and genetic counseling for this family.
Humans ; Child ; Classical Lissencephalies and Subcortical Band Heterotopias ; Exons ; Computational Biology ; Genetic Counseling ; Genitalia ; Transcription Factors ; Homeodomain Proteins

Objective:To analyse the current status of parturients' readiness for discharge, family functioning and psychological consistency, and to explore the relationship readiness for discharge, family functioning and psychological consistency.Methods:This was a cross-sectional study. Using the convenient sampling method, a total of 429 parturient women admitted to Obstetrics Department in two ClassⅢ Grade A hospitals in Yinchuan City from March to July 2022 were selected as the research objects. General Information Questionnaire, Family Assessment Device (FAD) , Readiness for Hospital Discharge Study-New Mother Form (RHDS-NMF) and Sense of Coherence-13 (SOC-13) were used to investigate the patients. A total of 429 questionnaires were sent out in this study, and 418 questionnaires were effectively collected, with an effective recovery rate of 97.44% (418/429) .Results:The total score of RHDS-NMF in 418 parturient women was (131.18±24.96) , the total score of FAD was (127.76±15.57) , and the total score of SOC-13 was (54.59±7.22) . The discharge readiness of parturient women was negatively correlated with family function ( r=-0.332, P<0.01) , while discharge readiness was positively correlated with psychological consistency ( r=0.253, P<0.01) . The mediation effect analysis results showed that psychological consistency played a partial mediating role between family function and discharge readiness in parturient women, with a mediation effect value of -1.105 ( P<0.05) , accounting for 27.1% of the total effect. Conclusions:Psychological consistency plays a partial mediating role between family function and discharge readiness in parturient women. Medical staff should pay attention to the evaluation and intervention of parturients' psychological consistency, enhance their level of psychological consistency and improve readiness for discharge.

Objective:To evaluate the effects of comprehensive rehabilitation nursing based on guided education and training on rehabilitation treatment in school-age children with drug-resistant epilepsy undergoing epileptogenic focus resection.Methods:Fifty movement disorders school-age children with drug-resistant epilepsy undergoing epileptogenic focus resection in Institute of Functional Neurosurgery, Xuanwu Hospital, Capital Medical University from January 2017 to December 2019 were enrolled in this study. The child patients were divided into the control group and the observation group according to the order of admission, with 25 cases in each group, and the control group was from January 2017 to June 2018 while the observation group was from July 2018 to December 2019. In the control group, routine nursing after epileptogenic focus resection and functional exercise were conducted. The observation group was treated with comprehensive rehabilitation nursing based on guided education and training. On the third day after operation, the day of discharge respectively, the motor function and activity of daily living of the two groups were evaluated. The satisfaction of children′s parents on nursing was compared between the two groups.Results:The scores of motor function on the third day after operation and the day of discharge were (57.0 ± 6.8), (73.0 ± 5.4) points respectively in the observation group, and those in the control group were (55.0 ± 5.6), (65.6 ± 5.9) points. There were significant differences intra group comparison ( t=-9.26, -6.48, both P<0.05). The activity of daily living scores on the third day after operation and the day of discharge were (45.2 ± 5.9), (74.2 ± 8.3) points respectively in the observation group, and those in the control group were (44.0 ± 5.8), (60.2 ± 7.6) points. There were significant differences intra group comparison ( t=-14.33, -8.51, both P<0.05). There were significant differences between the scores of motor function and the ability of daily living scores on the day of discharge in the observation group and the control group ( t values were -4.65, -6.25, both P<0.05). The satisfaction of children′s parents on nursing in the observation group and the control group were 96.0% (24/25), 72.0% (18/25) respectively. There was significant difference between the two groups ( χ 2=8.78, P<0.05). Conclusions:Rehabilitation nursing based on guided education and training can improve motor function and ability of daily living in movement disorders school-age children with drug-resistant epilepsy undergoing epileptogenic focus resection, promote the functional recovery of children after surgery and increase the satisfaction of children′s parents on nursing.