Objective To express the gene encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase (Sj32) and evaluate the potential of the recombinant protein rSj32 in diagnosis of domestic animal schistosomiasis. Methods The DNA fragment encoding mature protease of Schistosoma japonicum asparaginyl endopeptidase was cloned with PCR from pET-28(a)/Sj32, and a recombinant plasmid was previously constructed in the laboratory, which contained the ORF of the gene encoding the pro-enzyme Sj32. The amplified DNA fragment was subcloned into pET-28a( + ) and the recombinant plasmid was transformed into E. coli BL21 (DE3) to express the mature protease of Sj32. Then the recombinant antigen (rSj32) was used in ELISA assay to diagnose schistosomiasis of mice, rabbits and water buffalo artificially infected. The detection effects of soluble Schistosoma japonicum egg antigen (SEA) , rSj32 and the recombinant 23 KDa membrane protein were compared. Results The recombinant antigen rSj32 with a molecular weight 41 KDa was successfully produced in E. coli BL21 ( DE3) and was purified with His Column with a yield of 25 mg/L E. coli culture. By using rSj32 as coating antigen in ELISA assay to detect the specific antibody in artificially infected mice, rabbits and buffalo, the sensitivities were 88.9% , 85.0% and 71.8% , respectively, the specificities were 100% , 96.7% and 96. 9% , respectively. There were no significant differences among the detection results of rSj32, SEA and rSj23. Conclusion rSj32 is a promising antigen for serological diagnosis of domestic animal schistosomiasis.

OBJECTIVE: To explore a rare anatomic malformation of ethmoid and its clinical features, as well as an effective way of treatment. METHOD: Four cases from 2000-2009 of the first affiliated hospital of china medical university were studied according to the symptom,CT scanning, pathological examination and treatment process, respectively. RESULT: Ethmoid bone gasification complicated with infection resulted in a rare anatomic and pathological disorder. The symptom and treatment were largely dependent on the size of gasification and degree of infection. CONCLUSION CT Scanning plays a critical role in the diagnosis. In most case,a surgical treatment under endoscope is generally applied as a suitable way for clinical therapy. The regular reexamination is recommended for preventing recurrence.
Adolescent ; Bone Cysts ; complications ; diagnosis ; surgery ; Child ; Child, Preschool ; Ethmoid Bone ; abnormalities ; pathology ; Female ; Humans ; Male ; Tomography, X-Ray Computed

Objective:To study the distribution of ischemic stroke treatment with data mining technology and evaluate its clinical efficacy. Method:China National Knowledge Infrastructure Database（CNKI），China Science and Technology Reader's Digest Database（VIP），Wanfang Data，Chinese Biomedical Literature Database（Sino Med）were retrieved from January 1978 to December 2018. The clinical observation and study literatures on the treatment of ischemic stroke with the combination of traditional Chinese medicine and Western medicine were retrieved in the four databases. After standardized and hierarchical collection and processing of all syndromes，treatment methods，prescriptions and other information in the literatures，a database of syndrome elements and treatment of ischemic stroke was established. Syndrome factors and treatment methods were analyzed by scale evaluation and hierarchical classification methods. Kendall's tau-b correlation analysis，principal component analysis and other statistical methods were used to describe the correlation and distribution of syndrome factors and treatment methods of ischemic stroke. Result:The results of heterogeneity analysis showed that the included literatures were homogeneous and could be combined with subsequent statistics. A total of 450 syndromes and treatment methods were included in this study，and 1 287 single syndrome elements and 1 562 single treatment methods were obtained after unified and standardized splitting. Besides the corresponding syndrome elements and treatment methods，phlegm-dampness-invigorating Qi（-0.52） and Qi deficiency-invigorating Qi（-0.56） were also highly correlated. The study team represented the importance of syndrome and treatment elements with class Ⅰ，Ⅱ，Ⅲ from high to low. Qi deficiency，blood stasis and fire heat，phlegm，viscera excess were class Ⅰ syndrome elements；Yin deficiency，endogenous wind were class Ⅱ syndrome elements；Yin deficiency and Yang deficiency were class Ⅲ syndrome elements；Removing phlegm dampness，clearing heat，clearing the hollow viscera and extinguishing wind，promoting blood circulation to remove blood stasis，tonifying Qi were class Ⅰ treatment of ischemic stroke，and removing phlegm dampness，clearing heat，clearing the hollow viscera were more likely to appear simultaneously； and extinguishing wind，activating blood circulation and removing blood stasis，and benefiting Qi were more likely to appear simultaneously. Nourishing Yin and regulating Qi were class Ⅱ therapies of ischemic stroke，which were highly correlated and often appear simultaneously. Inducing resuscitation，tonifying Yang and dredging collaterals were class Ⅲ，Ⅳ，Ⅴ therapies. Conclusion:Qi deficiency，blood stasis，phlegm dampness，fire heat and viscera excess were the main syndromes of ischemic stroke，while Qi deficiency and blood stasis，phlegm heat and viscera excess were the main syndromes. Eliminating phlegm and dampness，clearing heat，clearing the hollow viscera，promoting blood circulation and removing blood stasis，extinguishing wind and benefiting Qi were the main therapies for the treatment of ischemic stroke. In clinical treatment for ischemic stroke，the therapies for relieving phlegm and dampness，clearing heat and relieving organs are often used in combination，and the therapies for promoting blood circulation and removing blood stasis were often used in combination with the therapies for invigorating Qi and extinguishing wind for the synergistic effect.

Schizophrenia is a serious mental disease. The diagnosis of schizophrenia so far relies heavily on subjective evidence, including self-reported experiences by patients, manifestations described by relatives, and abnormal behaviors assessed by psychiatrists. The diagnosis, monitoring of the disease progression and therapy efficacy assessment are challenging due to the lack of established laboratory biomarkers. Based on the current literature, clinical consensus, guidelines, and expert recommendations, this review highlighted evidence-based potential laboratory biomarkers for the diagnosis of schizophrenia, including genetic biomarkers, neurotransmitters, neurodevelopmental-related proteins, and intestinal flora, and discussed the potential future directions for the application of these biomarkers in this field, aiming to provide an objective basis for the use of these biomarkers in the early and accurate diagnosis, treatment, and prognosis and rehabilitation assessment of schizophrenia.

Objective: To investigate the role and mechanism of Vγ4 T cells in impaired wound healing of rapamycin-induced full-thickness skin defects in mice. Methods: The experimental research methods were applied. Eighty-six C57BL/6J male mice (hereinafter briefly referred to as wild-type mice) aged 8-12 weeks were selected for the following experiments. Vγ4 T cells were isolated from axillary lymph nodes of five wild-type mice for the following experiments. Intraperitoneal injection of rapamycin for 42 mice was performed to establish rapamycin-treated mice model for the following experiments. Eighteen wild-type mice were divided into normal control group without any treatment, trauma only group, and trauma+CC chemokine ligand 20 (CCL20) inhibitor group according to the random number table (the same grouping method below), with 6 mice in each group. The full-thickness skin defect wound was made on the back of mice in the latter two groups (the same wound model below), and mice in trauma+CCL20 inhibitor group were continuously injected subcutaneously with CCL20 inhibitor at the wound edge for 3 days after injury. Another 6 rapamycin-treated mice were used to establish wound model as rapamycin+trauma group. On post injury day (PID) 3, the epidermal cells of the skin tissue around the wound of each trauma mice were extracted by enzyme digestion, and the percentage of Vγ4 T cells in the epidermal cells was detected by flow cytometry. In normal control group, the epidermal cells of the normal skin tissue in the back of mice were taken at the appropriate time point for detection as above. Five wild-type mice were used to establish wound models. On PID 3, the epidermal cells were extracted from the skin tissue around the wound. The cell populations were divided into Vγ4 T cells, Vγ3 T cells, and γδ negative cells by fluorescence-activated cell sorter, which were set as Vγ4 T cell group, Vγ3 T cell group, and γδ negative cell group (with cells in each group being mixed with B16 mouse melanoma cells), respectively. B16 mouse melanoma cells were used as melanoma cell control group. The expression of interleukin-22 (IL-22) mRNA in cells of each group was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction (RT-PCR), with the number of samples being 6. Thirty rapamycin-treated mice were used to establish wound models, which were divided into Vγ4 T cell only group and Vγ4 T cell+IL-22 inhibitor group performed with corresponding injections and rapamycin control group injected with phosphate buffer solution (PBS) immediately after injury, with 10 mice in each group. Another 10 wild-type mice were taken to establish wound models and injected with PBS as wild-type control group. Mice in each group were injected continuously for 6 days. The percentage of wound area of mice in the four groups was calculated on PID 1, 2, 3, 4, 5, and 6 after injection on the same day. Six wild-type mice and 6 rapamycin-treated mice were taken respectively to establish wound models as wild-type group and rapamycin group. On PID 3, the mRNA and protein expressions of IL-22 and CCL20 in the peri-wound epidermis tissue of mice in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively. The Vγ4 T cells were divided into normal control group without any treatment and rapamycin-treated rapamycin group. After being cultured for 24 hours, the mRNA and protein expressions of IL-22 of cells in the two groups were detected by real-time fluorescence quantitative RT-PCR and Western blotting, respectively, with the number of samples being 6. Data were statistically analyzed with independent sample t test, analysis of variance for repeated measurement, one-way analysis of variance, Bonferroni method, Kruskal-Wallis H test, and Wilcoxon rank sum test. Results: The percentage of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in trauma only group on PID 3 was 0.66% (0.52%, 0.81%), which was significantly higher than 0.09% (0.04%, 0.14%) in the epidermal cells of the normal skin tissue of mice in normal control group (Z=4.31, P<0.01). The percentages of Vγ4 T cells in the epidermal cells of the skin tissue around the wound of mice in rapamycin+trauma group and trauma+CCL20 inhibitor group on PID 3 were 0.25% (0.16%, 0.37%) and 0.24% (0.17%, 0.35%), respectively, which were significantly lower than that in trauma only group (with Z values of 2.27 and 2.25, respectively, P<0.05). The mRNA expression level of IL-22 of cells in Vγ4 T cell group was significantly higher than that in Vγ3 T cell group, γδ negative cell group, and melanoma cell control group (with Z values of 2.96, 2.45, and 3.41, respectively, P<0.05 or P<0.01). Compared with that in wild-type control group, the percentage of wound area of mice in rapamycin control group increased significantly on PID 1-6 (P<0.01), the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 1 and PID 3-6 (P<0.05 or P<0.01). Compared with that in rapamycin control group, the percentage of wound area of mice in Vγ4 T cell only group decreased significantly on PID 1-6 (P<0.05 or P<0.01). Compared with that in Vγ4 T cell only group, the percentage of wound area of mice in Vγ4 T cell+IL-22 inhibitor group increased significantly on PID 3-6 (P<0.05 or P<0.01). On PID 3, compared with those in wild-type group, the expression levels of IL-22 protein and mRNA (with t values of -7.82 and -5.04, respectively, P<0.01) and CCL20 protein and mRNA (with t values of -7.12 and -5.73, respectively, P<0.01) were decreased significantly in the peri-wound epidermis tissue of mice in rapamycin group. After being cultured for 24 hours, the expression levels of IL-22 protein and mRNA in Vγ4 T cells in rapamycin group were significantly lower than those in normal control group (with t values of -7.75 and -6.04, respectively, P<0.01). Conclusions: In mice with full-thickness skin defects, rapamycin may impair the CCL20 chemotactic system by inhibiting the expression of CCL20, leading to a decrease in the recruitment of Vγ4 T cells to the epidermis, and at the same time inhibit the secretion of IL-22 by Vγ4 T cells, thereby slowing the wound healing rate.
Animals ; Male ; Melanoma ; Mice ; Mice, Inbred C57BL ; RNA, Messenger ; Sirolimus/pharmacology* ; T-Lymphocytes ; Wound Healing

The effect of porcine SIRT5 on replication of foot and mouth disease virus type O(FMDV-O)and the underlying regulatory mechanism were investigated.Western blot and RT-qPCR analyses were employed to monitor expression of endoge-nous SIRT5 in PK-15 cells infected with FMDV-O.Three pairs of SIRT5-specific siRNAs were synthesized.Changes to SIRT5 and FMDV-O protein and transcript levels,in addition to virus copy numbers,were measured by western blot and RT-qPCR analyses.PK-15 cells were transfected with a eukaryotic SIRT5 expression plasmid.Western blot and RT-qPCR analyses were used to explore the impact of SIRT5 overexpression on FMDV-O replication.Meanwhile,RT-qPCR analysis was used to detect the effect of SIRT5 overexpression on the mRNA expression levels of type I interferon-stimulated genes induced by SeV and FMDV-O.The results showed that expression of SIRT5 was up-regulated in PK-15 cells infected with FMDV-O and siRNA interfered with SIRT5 to inhibit FMDV-O replication.SIRT5 overexpression promoted FMDV-O replication.SIRT5 over-expression decreased mRNA expression levels of interferon-stimulated genes induced by SeV and FMDV-O.These results suggest that FMDV-O infection stimulated expression of SIRT5 in PK-15 cells,while SIRT5 promoted FMDV-O rep-lication by inhibiting production of type I interferon-stimula-ted genes.These findings provide a reference to further ex-plore the mechanism underlying the ability of porcine SIRT5 to promote FMDV-O replication.

Enhanced recovery after surgery (ERAS) measures have not been systematically applied in transurethral surgery for benign prostatic hyperplasia (BPH). This study was performed on patients with BPH who required surgical intervention. From July 2019 to June 2020, the ERAS program was applied to 248 patients, and the conventional program was applied to 238 patients. After 1 year of follow-up, the differences between the ERAS group and the conventional group were evaluated. The ERAS group had a shorter time of urinary catheterization compared with the conventional group (mean ± standard deviation [s.d.]: 1.0 ± 0.4 days vs 2.7 ± 0.8 days, P < 0.01), and the pain (mean ± s.d.) was significantly reduced through postoperative hospitalization days (PODs) 0-2 (POD 0: 1.7 ± 0.8 vs 2.4 ± 1.0, P < 0.01; POD 1: 1.6 ± 0.9 vs 3.5 ± 1.3, P < 0.01; POD 2: 1.2 ± 0.7 vs 3.0 ± 1.3, P < 0.01). No statistically significant difference was found in the rate of postoperative complications, such as postoperative bleeding (P = 0.79), urinary retention (P = 0.40), fever (P = 0.55), and readmission (P = 0.71). The hospitalization cost of the ERAS group was similar to that of the conventional group (mean ± s.d.: 16 927.8 ± 5808.1 Chinese Yuan [CNY] vs 17 044.1 ± 5830.7 CNY, P =0.85). The International Prostate Symptom Scores (IPSS) and quality of life (QoL) scores in the two groups were also similar when compared at 1 month, 3 months, 6 months, and 12 months after discharge. The ERAS program we conducted was safe, repeatable, and efficient. In conclusion, patients undergoing the ERAS program experienced less postoperative stress than those undergoing the conventional program.
Male ; Humans ; Prostatic Hyperplasia/complications* ; Quality of Life ; Transurethral Resection of Prostate/adverse effects* ; Treatment Outcome ; Enhanced Recovery After Surgery

OBJECTIVE: Dexmedetomidine (DEX), the most specific α ₂-adrenergic receptor agonist widely used for its sedative and analgesic properties, has been reported to upregulate HIF-1α expression to protect hypoxic and ischemic tissues. However, it is largely unclear whether DEX can also upregulate Hypoxia-inducible factor-1 alpha (HIF-1α) expression and its downstream vascular endothelial growth factor-A (VEGFA) in cancer tissues with oxygen-deficient tumor microenvironment. METHODS: We used SMMC-7721 cells, MHCC97-H cells, and a mouse model of orthotopic hepatic carcinoma to explore the effect of DEX on angiogenesis and vasculogenic mimicry (VM) and its mechanism. Under normoxic (20% O ₂) and hypoxic (1% O ₂) conditions, DEX was used to intervene cells, and yohimbine was used to rescue them. RESULTS: The results showed that DEX promoted angiogenesis and VM in human liver cancer cells within a certain dose range, and the addition of yohimbine inhibited this effect. DEX could activate HIF-1α/VEGFA pathway, which was further verified by silencing HIF-1α. Consistently, in vivo results also showed that DEX can up-regulate HIF-1α/VEGFA expression, and enhance the number of VM channels and microvessel density (MVD). CONCLUSION We believe that HIF-1α/VEGFA might be an important signaling pathway by which DEX promotes angiogenesis and VM formation in human hepatocellular carcinoma, whereas α ₂-adrenergic receptor mediation might be the critical mechanisms.
Animals ; Humans ; Mice ; Adrenergic alpha-2 Receptor Agonists/pharmacology* ; Carcinoma, Hepatocellular ; Cardiovascular Physiological Phenomena ; Dexmedetomidine/pharmacology* ; Hypoxia ; Liver Neoplasms/drug therapy* ; Oxygen ; Tumor Microenvironment ; Vascular Endothelial Growth Factor A/genetics* ; Receptors, Adrenergic, alpha-2/metabolism*