Objective To clone the full-length sequence,subcellular localization,and analyze the expression patterns of the β-caryophyllene synthase gene(AaCPS)of Artemisia argyi,in order to lay a foundation for the gene function analysis of sesquiterpene biosynthesis pathway in A.argyi.Methods The genes annotated as CPS were selected from the transcriptome data of A.argyi.The full-length cDNA sequence of the target gene was obtained by PCR,and the coding region was analyzed using bioinformatics.A prokaryotic expression vector was constructed and transformed into Escherichia coli competent cells to express recombinant protein.A green fluorescent protein(GFP)fused expression vector was constructed,and the subcellular localization of AaCPS was observed using Agrobacterium tumefaciens transient expression method in tobacco.Real-time quantitative PCR(qRT-PCR)was used to analyze the expression patterns of the AaCPS at different harvest times(April,May,June,July).Determination of β-caryophyllene by HS-SPME-GC-MS and correlation analysis.Results The AaCPS gene was cloned from A.argyi.The full-length open reading frame(ORF)was 1647 bp,encoding 548 amino acids.The molecular weight was 63 667 Da with a theoretical pI of 5.94.AaCPS contained conserved Terpen_synth and Terpen_synth_C domains.Phylogenetic analysis indicated that AaCPS was closely related to the CPS protein of Artemisia annua.SDS-PAGE showed the presence of target protein bands between 75 000-120 000 Da(containing 28 132 Da of the labeled protein),indicating successful expression of the AaCPS protein.The protein was found to be localized in the cytoplasm.As shown by qRT-PCR,the expression of AaCPS gene showed an upward trend,reaching the highest in July.The results of HS-SPME-GC-MS showed that the content of β-caryophyllene increased gradually from April to the highest in June.It is consistent with the trend of AaCPS gene expression from April to June.Conclusion Through the cloning and analysis of AaCPS gene,a foundation has been laid for further study of the functional identification of the AaCPS gene in the sesquiterpenes biosynthesis pathway in A.argyi.

Objective:To observe the therapeutic effect of autologous neurosensory retinal transplantation in repairing unhealed giant macular hole after pars plana vitrectomy (PPV).Methods:A prospective clinical study. From July 2022 to December 2023, 12 patients (12 eyes) with refractory large macular hole who received autologous neurosensory retinal transplantation treatment in Department of Ophthalmology of the First Affiliated Hospital of Zhengzhou University were selected for the study. The macular hole in affected eyes still did not close after PPV combined with inner limiting membrane removal or tamponade, and the diameter of macular hole were greater than 600 μm. All affected eyes received best corrected visual acuity (BCVA) and optical coherence tomography (OCT) examinations. The BCVA examination employed the international standard visual acuity chart, with results converted to logarithm of the minimum angle of resolution (logMAR) visual acuity for statistical analysis. During the surgery, a piece of healthy retinal neuroepithelial tissue, approximately 0.3 optic disc diameters larger than the macular hole, was removed from the upper retinal periphery and used as a graft. The graft was inserted into the macular hole with the aid of intraoperative OCT. Post-surgery, the vitreous cavity was filled with silicone oil or sterile air. The follow-up period after surgery was 6 months. The thickness of the retinal grafts was measured using the same equipment as before surgery at 3 days, 1, 3, and 6 months post-surgery. The primary focus was on observing the macular hole closure rate and changes in BCVA at 6 months post-operation. A paired t-test was used to compare BCVA before and after surgery. Results:In the sample of 12 cases (12 eyes), there were 5 males with 5 eyes and 7 females with 7 eyes. The mean age was (50.4±12.6) years. The mean macular hole diameter was (1 085.6±344.0) μm; The mean eye axis length was (27.64±4.19) mm. At 6 months after surgery, all affected eyes showed macular hole were completely closed (100.0%, 12/12). The thickness of the retinal graft was measured as (206.8±21.0), (170.8±23.3), (165.6±31.6), and (157.9±31.1) μm at 3 days, 1, 3, and 6 months post-surgery, respectively. At before and 6 months after surgery, the logMAR BCVA of the affected eyes was 1.28±0.39 and 0.95±0.22, respectively. The difference in logMAR BCVA before and after surgery was statistically significant ( t=3.40, P<0.05). Conclusion:Autologous neurosensory retinal transplantation could effectively improve the closure rate of refractory large macular hole and improve or stabilize vision in the short run.

Objective:To observe the histopathological changes in peripheral retinal lesions under intraoperative optical coherence tomography (iOCT).Methods:A retrospective case series study. Eighty-eight patients (194 eyes) who underwent vitreoretinal surgery in the Department of Ophthalmology at the East Ward of the First Affiliated Hospital of Zhengzhou University from October 2021 to May 2022 in 94 eyes were included in the study. Among them, 49 cases were male and 39 cases were female, with the mean age of (50.93±17.55) years. Ninety-four eyes included 32 eyes with retinal detachment, 6 eyes with proliferative diabetic retinopathy, 28 eyes with vitreous hemorrhage, 8 eyes with ocular trauma, 14 eyes with the macular lesion, 1 eye with uveitis, 1 eye with family exudative vitreoretinopathy (FEVR), 1 eye with acute retinal necrosis (ARN), and 3 eyes with lens dislocation. All affected eyes were examined with iOCT during vitreoretinal surgery. The iOCT scanning of the peripheral retina was performed with the help of episcleral pressure. The pre-equatorial and serrated edge anterior and posterior of retinas were scanned according to the characteristics of different fundus diseases. Various abnormal fundus manifestations were recorded.Results:In 94 eyes, 53 eyes (56.38%, 53/94) have different types of retinopathy in the peripheral retina. Of these, 7 eyes (7.45%) have retinal cystoid degeneration; 19 eyes (20.21%) have lattice degeneration; and 8 eyes (8.51%) have pigment degeneration; 9 eyes (9.57%) have pavement-like degeneration; 7 eyes (7.45%) have small occult holes; 1 eye (1.06%) has familial exudative vitreoretinopathy (FEVR) serrated edge "dyke-like" proliferative degeneration; 4 eyes (4.26%) have vitreous and retinopathy adhesions; and one eye (1.06%) has ARN.Conclusion:With clear refractive media, iOCT can provide clear scans of different peripheral retinal lesions.

Objective:To compare the dosimetric differences between volumetric-modulated arc therapy (VMAT) and intensity-modulated radiation therapy (IMRT) on planning target volume (PTV) and organ-at-risk (OAR) for breast cancer after modified radical mastectomy, aiming to provide evidence-based reference for clinical practice.Methods:According to strict inclusion and exclusion criteria, literature search was performed in PubMed, Cochrane Library, FMRS, CNKI, Wanfang Data and VIP full text databases from the inception of databases up to March 2020. The controlled clinical trials of dosimetric comparison between VMAT and IMRT for breast cancer following modified radical mastectomy were selected. The meta-analysis was performed using Stata14 software.Results:The meta-analysis included 281 patients from 13 observational studies. Compared with IMRT, VMAT significantly increased the PTV dose coverage D 95%( P<0.001) and significantly improved the PTV homogeneity index (HI, P<0.001) and conformity index (CI, P=0.004). Compared with IMRT, VMAT more effectively decreased the ipsilateral lung V 20Gy (WMD=1.332, P=0.027) and contralateral lung V 10Gy ( P=0.003). There were no significant differences in theD mean, V 5Gy, V 10Gy and V 30Gy of the ipsilateral lung, D mean and V 5Gy of the contralateral lung, D mean, V 10Gy and V 30Gy of the heart between VMAT and IMRT (all P>0.05). Compared with VMAT, IMRT reduced the cardiac V 5Gy ( P=0.001). However, sensitivity analysis of included literature on cardiac V 5Gy showed that the P value was reversed, indicating that the stability of the results was poor. VMAT significantly shortened the delivery time ( P<0.001) and the number of monitor units ( P<0.001) compared to IMRT. Conclusion:Compared with IMRT, VMAT can achieves superior target dose coverage, HI and CI, better protection for the ipsilateral and contralateral lung, fewer monitor units and shorter delivery time.

Objective:To master the changes of Kashin-Beck disease and the examination and acceptance in Zhalantun City, and to provide scientific basis for formulating prevention and control strategies of Kashin-Beck disease.Methods:From 2016 to 2017, in each county under the jurisdiction of Zhalantun City, 5 townships affected by the disease were selected, 3 villages were selected from each township, and the prevalence of Kashin-Beck disease of all residential children aged 7 to 12 years who lived in the villages for more than 6 months were investigated. The clinical and X-ray diagnosis were performed according to the "Diagnosis of Kashin-Beck Disease" (WS/T 207-2010) standard, and the prevalence of Kashin-Beck disease in children aged 7 to 12 years in Zhalantun City in 2016 and 2017 were compared. In accordance with the "Key Endemic Disease Control and Elimination Evaluation Measures" ([2014]79), the condition of Kashin-Beck disease and the implementation of its prevention and control measures was assessed.Results:From 2016 to 2017, 1 697 children aged 7 to 12 years were examined, there were no clinical cases of Kashin-Beck disease and 11 cases of X-ray positive changes. Among them, 844 children were examined in 2016, the positive rate of X-ray was 0.24% (2/844), the positive rate of metaphyseal was 0.24% (2/844), and no positive changes of extremities and triad were detected. A total of 853 children were examined in 2017, the positive rate of X-ray was 1.06% (9/853), the positive rate of metaphyseal was 1.06% (9/853), and no positive changes of extremities and triad were detected. The positive rate of X-ray and metaphyseal of Kashin-Beck disease in children aged 7 - 12 years in 2017 were higher than those in 2016 (χ 2 = 4.409, 4.409, P < 0.05). All surveyed villages had reached the national elimination standard (no clinical cases for children aged 7 - 12 years, X-ray positive rate ≤3% and no cases of hand bone end changes); the organization management scores of Kashin-Beck disease prevention and treatment in Zhalantun City in 2016 and 2017 were 88 points, reaching the qualified standard ( > 85 points). Conclusion:The condition of Kashin-Beck disease in Zhalantun City has reached the standard of elimination, which lays a foundation for further comprehensive elimination of Kaschin-Beck disease in Hulunbuir City.

Objective: To explore the safety and feasibility of transumbilical single channel laparoscopic cryptorchidopexy with conventional instruments in treatment of cryptorchidism in children. Methods: Thirty-five children with cryptorchidism were treated by transumbilical single channel laparoscopic cryptorchidopexy with conventional instruments from November 2017 to March 2018 at Department of Pediatric Surgery, the First Affiliated Hospital of Guangxi Medical University.Clinical data were recorded including the general information, operation time, intraoperative blood loss, postoperative testicular position and postoperative complications, etc. Results: All 35 patients completed the cryptorchidopexy in one operation.Among those 35 patients, 28 patients had unilateral lesions and 7 patients had bilateral lesions, with a total of 42 sides.Twenty-six patients were of groin type(33 sides), 9 patients abdominal type(9 sides). The average operation time was (40.4±8.2) min for unilateral lesion, and (62.5±15.3) min for bilateral lesions; the average intraoperative blood loss was (3.0±1.8) mL; and the average postoperative hospital stay was (6.4±2.1) h. Testis of 32 patients (40 sides) were fixed in the middle and bottoms of the scrotum, and testis of 2 patients (2 sides) were fixed on the top of the scrotum.One case had poor healing of naval incision, which was recovered in 1 week after dressing change.There were no postoperative complications such as scrotum hematoma, infection of incision, testicular epididymitis, testicular necrosis, testicular retraction or testicular atrophy, etc.No obvious visible scar was found in the abdomen review 2 weeks postoperatively, with the incidence of complications was 2.9%(1/35 cases). Conclusions Transumbilical single channel laparoscopic cryptorchidopexy with conventional instruments in treatment of cryptorchidism is characterized by small trauma, quick recovery, low complication rate and visible scar free result, which is worthy of clinical promotion, but the operation is relatively complex and the correct procedures are required.

Objective: To forecast the risk distribution of inter-animal plague in Meriones unguiculatus effectively and provide scientific evidence for prevention and control of inter-animal plague, through studying the correlation between meteorological and environmental factors and inter-animal plague in Meriones unguiculatus. Methods: Positive data of plague bacterial culture in 30 epidemic source areas of Meriones unguiculatus in the Inner Mongolia plateau from 2005 to 2018, including detecting time, number of bacteria, latitude and longitude or detailed location, host type, were from the Chinese Disease Prevention and Control System Plague Prevention Management Information System and related professional institutions for plague prevention and treatment. Logistic regression was used to explore the relationship between the inter-animal plague and climate-related risk factors. The Maximum Entropy (Maxent) model was used to predict the habitat distributions of inter-animal plague, and the receiver operating characteristic (ROC) curve was used to validate the model. Results: There were 11 climatic factors including annual mean temperature, isothermality, temperature seasonality, mean temperature of driest quarter, mean temperature of warmest quarter, mean temperature of coldest quarter, annual precipitation, precipitation seasonality, globcover, normalized difference vegetation index and slope, were related to the outbreak of plague among Meriones unguiculatus and included in the model (OR = 1.302, 0.455, 0.957, 0.930, 4.864, 0.179, 0.986, 1.126, 0.992, 0.981, 0.721, P < 0.01). The increase of annual mean temperature, mean temperature of warmest quarter and precipitation seasonality will increase the risk of animal plague in the plague foci of Meriones unguiculatus; the increase of isothermality, temperature seasonality, mean temperature of driest quarter, mean temperature of coldest quarter, annual precipitation, globcover, normalized difference vegetation index, and slope will reduce the risk of animal plague in the plague foci of Meriones unguiculatus. The areas under the curve (AUCs) of the Maxent model training data and test data were 0.988 and 0.985, the prediction effect of the model was better. The habitat distribution of Meriones unguiculatus plague mainly concentrated in the central and northern Ulanqab plateau, Ordos plateau, and eastern Hetao plain. Conclusions The use of Maxent model and climate data can predict the potential risks and spatial distribution of animal plague in Meriones unguiculatus; the results are accurate and reliable.

Objective:To detect the expression and clinical significance of β-tubulin Ⅲ in cancer tissue of the patients with colon adenocarcinoma,and to explore its clinical significance.Methods:A total of 111 colon adenocarcinoma tissue samples were obtained.According to the location of β-tubulin Ⅲ positive cells, all patients were divided into front group (n=72) and non-front group (n=39).The positive expression rate of β-tubulin Ⅲ in the patients with colon adenocarcinoma was detected with immunohistochemistry.The correlations among the expression of β-tubulin Ⅲ and gender,age,tumor differentiation, clinical stage, lymph node metastasis, recurrence and death were analyzed.Results: The expression levels of β-tubulin Ⅲ had no significant differences between the patients with different gerder,age,lymph node metastasis,clinical stages,death and recurrence.The positive expression rates of β-tubulin in cancer tissue of the patients had significant difference between front and non-front groups (χ2=8.76, P=0.01).Lowly-to-moderately differentiated tissue was more common in front group, and highly-differentiated tissue was more common in non-front group(χ2=6.88, P=0.03).There were significant differences in the expression levels of β-tubulin Ⅲ between cancer tissues with different differentiation degrees (χ2=5.74, P=0.04).In non-front group, lymph node metastasis was closely correlated with the expression of β-tubulin Ⅲ (χ2=6.02,P=0.05).The results of immunohistochemical staining showed that the β-tubulin Ⅲ positive-expressing cells were colored brown-yellow.The number of cells with positive β-tubulin Ⅲ expression was significantly increased in highly differentiated tissue compared with low-differentiated tissue.Conclusion:The expression of β-tubulin Ⅲ is closely related to tumor differentiation in colon adenocarcinoma tissue.The highly differentiated colon adenocarcinoma tissue is more common in non-front group in which the expression of β-tubulin Ⅲ is related to lymph node metastasis.

Objective To investigate effects of polyphylin Ⅰ on the proliferation and apoptosis of human melanoma cell line A375,and to explore their mechanisms.Methods Normal human melanocytes isolated from healthy human foreskin were divided into 6 groups to be treated with 0,1.5,3.0,6.0,9.0,12.0 mg/L polyphyllin Ⅰ respectively.A375 melanoma cells were divided into 4 groups,i.e.,control group,1.5-,3.0-,6.0-mg/L polyphyllin Ⅰ groups,to be treated with 0,1.5,3.0,6.0 mg/L polyphyllin Ⅰ,respectively.Cell counting kit-8 (CCK8) assay was performed to evaluate the effect of polyphyllin Ⅰ on the proliferation of normal human melanocytes and A375 cells.Hoechst 33258 fluorescent staining was conducted to observe the morphology of apoptotic cells,flow cytometry to estimate cell cycle phase distribution and apoptosis rate,dichloro-dihydro-fluorescein diacetate (DCFH-DA) fluorescent probe assay to detect the level of reactive oxygen species (ROS),rhodamine-123 staining to evaluate changes of mitochondrial membrane potential,spectrophotography to detect the level of ATP in A375 cells,as well as levels of lactic acid and glucose in the culture supernatant of A375 cells,and Western blot analysis to determine the protein expression of Bcl-2,Bcl-2-related X protein (Bax),cleaved-caspase-3,cyclin D1 and pyruvate kinase isozyme type M2 (PKM2).Statistical analysis was carried out by using one-way analysis of variance (ANOVA) for comparisons among groups and Student-Newman-Keuls-q (SNK-q) test for multiple comparisons.Results CCK8 assay showed that the treatment with polyphyllin Ⅰ at concentrations of 1.5,3.0,6.0 mg/L for 48 hours had no effects on the proliferation of normal human melanocytes,but significantly inhibited the proliferation of A375 cells.The survival rate of A375 cells was significantly lower in the 1.5-,3.0-,6.0-mg/L polyphyllin Ⅰ groups than in the control group (P ＜ 0.01).After the treatment with polyphyllin Ⅰ,distinct apoptotic morphology of A375 cells was observed under fluorescence microscope.Additionally,along with the increase of polyphyllin Ⅰ concentrations (0,1.5,3.0,6.0 mg/L),there were gradual increasing trends in the apoptosis rate of A375 cells (4.25％ ± 1.27％,10.03％ ± 1.49％,36.62％ ± 1.97％,44.11％ ± 2.47％ respectively,F =665.7,P ＜ 0.01),the percentage of A375 cells at G0/G1 phase (54.13％ ± 2.57％,67.35％ ± 3.79％,74.39％ ± 3.29％,82.29％ ± 3.99％ respectively,F =71.81,P ＜ 0.01),the level of ROS in A375 cells (P ＜ 0.01),the level of glucose in the culture supernatant (P ＜ 0.01),and the protein expression of Bax and cleaved-caspase-3 (both P ＜ 0.01),while gradual decreasing trends were found in the levels of mitochondrial membrane potential and ATP in A375 cells (both P ＜ 0.01),the level of lactic acid in the culture supernatant (P ＜ 0.01),and the protein expression of Cyclin D1,Bcl-2 and PKM2 (all P ＜ 0.01).Conclusion Polyphyllin Ⅰ can effectively induce A375 cell apoptosis by promoting the production of ROS in A375 cells and decreasing the mitochondrial membrane potential,and arrest A375 cells at G0/G1 phase by inhibiting the expression of PKM2 and Cyclin D1.

Objective To explore the expression alteration and significance of inteleukin (IL)-37 in pso-riasis valguris (PV) patients. Methods Patients with PV had been treated with oral acitretin for 8 weeks. PASI score, ELISA and qRT-PCR were used to exam the data of 38 patients (PV group) and 32 controls (control group). Results IL-37 in PV group was significantly higher than that in control group (P < 0.001) and IL-37 changed slightly after 4 weeks of treatment(P > 0.05) but decreased obviously after 8 weeks(P < 0.001). Signif-icant correlations existed among PASI scores, IL-37, IFN-γ and IL-17, as well as among IL-37 and IFN-γ, IL-17 (P < 0.05). Conclusions The increase of IL-37 is relevant to PV development and is associated with pa-tients’ conditions, IFN-γ and IL-17 but the alteration of IL-37 is not related with IL-4.