Background Accurate measurement of corneal thickness is very important during the pre-and post-operative management of corneal surgical procedures,especially laser-assisted in-situ keratomileusis (LASIK),which is the most popular approach to the correction of refractive errors currently.This may be particularly important for the patients who have undergone previous laser refractive surgery with suboptimal outcomes and are being considered for an enhancement procedure.Objective This study was to compare the measuring outcomes of corneal thickness by slit-scanning pachymetry,non-contact specular microscope,anterior segment optical coherence tomography (AS-OCT)and ultrasound pachymetry,with a focus on central and midperipheral (from the central 3.0 mm) region of cornea in post-LASIK eyes.Methods Sixty-four right eyes of 64 patients who received LASIK were collected in Henan Eye Institute,Henan Eye Hospital from March to June 2011 with the equivalent spherical diopter of (-4.75±2.38)D and horizontal corneal diameter of (11.36±0.32)mm.Central corneal thickness was measured on each eye by using non-contact specular microscope (Topcon SP-3000P),slit-scanning pachymetry (Orbscan Ⅱ),AS-OCT and A-type ultrasound pachymetry,respectively,and the paracentral corneal thickness including 12:00,2:00,6:00 and 10:00 meridian was measured using Orbscan Ⅱ,non-contact specular microscope and AS-OCT.The measuring values and the agreement from different instruments were compared and evaluated.Results The mean central corneal thickness was (467.12±31.10)tμm for AS-OCT,(466.67±30.99)μm for ultrasound pachymetry,(441.84 ± 33.65) μm for specular microscopy and (422.51 ± 44.09) μm for Orbsan Ⅱ,respectively,showing a significant difference among the four methods (F =23.730,P =0.000).The central thickness value of the A-type ultrasound pachymetry was significantly higher than that of Orbsan Ⅱ or non-contact specular microscope (q =6.940,6.720,both at P =0.000).Compared with Orbscan Ⅱ,the measuring values of non-contact specular microscope and AS-OCT were significantly higher (q =-5.54,6.940,both at P =0.000),and the measuring value of AS-OCT was significantly higher that of non-contact specular microscope (q =6.800,P =0.000).The lowest difference value (25.3 μm)and the best agreement was found between the ultrasound pachymetry and AS-OCT.The paracentral corneal thickness values in 12:00,2:00,10:00,6:00 meridians were highest for Orbsan Ⅱ and the next for AS-OCT,and non-contact specular microscope had the lowest values,with significantly differences among them (F =5.020,22.950,67.890,18.850,all at P ＜ 0.01).Conclusions The corneal thickness values vary with the different instruments.Orbsan Ⅱ underestimates the central corneal thickness and overestimates the midperipheral corneal thickness,and non-contact specular microscope underestimates both the central and midperipheral corneal thickness.The measuring outcome from AS-OCT has a good agreement with ultrasound pachymetry and therefore they can be used interchangeably.

Objective We are going to establish a robust liquid chromatography-tandem mass spectrometric(LC-MS/MS) method for plasma aldosterone assay.Methods 324 healthy individuals were enrolled in Zhongshan Hospital from February to April in 2016 for reference interval survey.The signallinearity,lower limits of quantitation,precision and accuracy of LC-MS/MS have been evaluated.Results from LC-MS/MS and RIA methods were compared.Software SPSS17.0 software was used for statistical analysis.Results The performance characteristics for the method in terms of linearity,lowerlimits of quantitation,precision and accuracy were verified.Linear range of ALD were between 25-2000 pg/ml;the LC-MS/MS assay had a limit of quantitation of 20 pg/ml for ALD;the intra-and inter-assay CV of ALD were ＜10％ and ＜6％,respectively;the recovery of ALD from serum samples ranged between 97.3 and 105.8％ The reference value of ALD in health people ranged between 21-211.6 pg/ml The regression equation by LC-MS/MS (X) and RIA (Y) was:Y =0.271X + 138.900(r=0.43;n=322).Conclusion LC-MS/MS method is robust and reliable for the analysis of aldosterone in plasma and suitable for clinical application.

Objective To detect the genomic copy number variations(CNVs) of mice embryonic neural tissue with neural tube defects (NTDs) induced by methotrexate (MTX),and investigate the molecular genetic mechanisms between folic acid metabolism disorders and NTDs pathogenesis.Methods C57BL/6J NTD mice model was induced by MTX on gestational day 7.5,and the maternal serum and NTD embryonic neural tissues were collected;the array-comparative genomic hybridization(array-CGH) assay was utilized to analyze the whole genomic CNVs in NTD embryonic neural tissues;reverse transcription polymerase chain reaction(RT-PCR) was used to confirm the positive results;the maternal serum concentrations of folic acid and related metabolites and dihydrofolate reductase (DHFR) activity were detected by liquid chromatography-tandem mass spectrometry and enzymatic methods,respectively.Results ArrayCGH and RT-PCR results showed the 3 high confidence CNVs on XqE3,XqA1.1-qA2 and XqA1.1 in the NTD embryonic neural tissues.The NTD maternal serum concentrations of 5-methyltetrahydrofolate,5-formyltetrahydrofolic acid,S-adenosyl methionine and DHFR activity were reduced significantly compared with the control group,and there were statistical differences(all P ＜0.05).Conclusions There are obvious CNVs in embryonic neural tissue of NTD mice induced by MTX and folic acid dysmetabolism might cause mice embryonic neural tube developmental disorders through CNVs.

Objective To evaluate the efficacy of transarterial chemoembolization (TACE) and percutaneous cryosurgery sequential therapy for unresectable hepatocellular carcinoma (HCC).Methods Four hundred and twenty patients with unresectable HCC were divided into sequential TACE-cryosurgery sequential (sequential) group (n=290) and cryosurgery alone (cryoalone) group (n = 130). TACE was performed with the routine operation; the percutaneous cryosurgery was conducted 2 to 4 weeks after TACE. The patients were followed up at the first month and once every 2 to 3 month later. Liver ultrasound or both computer tomography and alpha fetal protein were examined during follow-up. Results During a mean follow-up of (42±17) months (range from 24 to 70 months), the local recurrence rate of ablated lesion was 17% for all the patients, 11% and 24% for patients in sequential group and cryoalone groups respectively (P=0. 001). The overall 1-, 2-, 3-, 4-and 5-year survival rate was 72%, 57%, 47%, 39% and 31%, respectively. The 1- and 2-year survival rates (71% and 61 % ) in sequential group were similar to those (73 % and 54 % ) in cryo-alone group (P=0.69 and 0. 147), while the 4- and 5-year survival rates were higher in sequential group (49 % and 39 % ) than those (29 % and 23 % ) in cryo-alone group (P= 0.001). Eighteen patients with large HCC (＞5 cm in diameter) in sequential group survived for more than 5 years while no one in cryo-alone group. Complication rate was 24% in all patients, 21% and 26% for the sequential and cryo-alone groups respectively (P=0. 06). The incidence of hepatic bleeding was higher in cryo-alone group than in sequential group (P=0. 02). Liver crack occurred in two patients of the cryoalone group. Conclusions Pre-cryosurgical TACE increased the cryoablation efficacy and decrease its complications, especially hepatic bleeding. TACE and cryosurgery sequential therapy may be a better treatment for unresectable HCC, especially for large HCC.

To identify novel transcripts and sRNA in genome of B .melitensis by transcriptome sequencing ,total RNA were extracted from B .melitensis culture and rRNA were removed .After the addition of adaptor ,RNA was reversely transcribed into cDNA ,which were then subjected to PCR amplification and sequencing .The generated reads were mapped to genome se‐quence of B .melitensis strain 16M .With the mapping results ,novel transcripts and sRNA were identified by bioinformatics methods .Sequencing results analysis showed that genome sequence was covered with the reads with good quality .A total of 773 genes were extended in their 5′and/or 3′ends of their original locations .Sixteen novel transcripts and 241 sRNAs candi‐dates were identified .RT‐PCR showed that some of the sRNAs were differentially expressed under stress conditions .In B . melitensis genome ,there is novel transcript which is not predicted .The sRNA does exist in B .melitensis and were expressed under different conditions .

Background and Objectives: Glioblastoma (GBM) is an aggressive primary brain tumor characterized by its hetero-geneity and high recurrence and lethality rates. Glioblastoma stem cells (GSCs) play a crucial role in therapy resistance and tumor recurrence. Therefore, targeting GSCs is a key objective in developing effective treatments for GBM. The role of Parathyroid hormone-related peptide (PTHrP) in GBM and its impact on GSCs remains unclear. This study aimed to investigate the effect of PTHrP on GSCs and its potential as a therapeutic target for GBM. Methods: and Results: Using the Cancer Genome Atlas (TCGA) database, we found higher expression of PTHrP in GBM, which correlated inversely with survival. GSCs were established from three human GBM samples obtained after surgical resection. Exposure to recombinant human PTHrP protein (rPTHrP) at different concentrations significantly enhanced GSCs viability. Knockdown of PTHrP using target-specific siRNA (siPTHrP) inhibited tumorsphere formation and reduced the number of BrdU-positive cells. In an orthotopic xenograft mouse model, suppression of PTHrP expression led to significant inhibition of tumor growth. The addition of rPTHrP in the growth medium counteracted the antiproliferative effect of siPTHrP. Further investigation revealed that PTHrP increased cAMP concentration and activated the PKA signaling pathway. Treatment with forskolin, an adenylyl cyclase activator, nullified the antiproliferative effect of siPTHrP. Conclusions Our findings demonstrate that PTHrP promotes the proliferation of patient-derived GSCs by activating the cAMP/PKA signaling pathway. These results uncover a novel role for PTHrP and suggest its potential as a therapeutic target for GBM treatment.

Objective To observe the clinical effect of the Bushen-Qiangjin capsule and calcium D in the treatment of aromatase inhibitors-associated bone loss (AIBL) in breast cancer patients. Methods A total of 65 patients were randomized into a treatment group and a control group, 32 in the control group taking calcium D, and 33 in the treatment group taking calcium D and Bushen-Qiangjin capsule. After a 3-month treatment, the bone mineral density T (BMD), bone-specific alkaline phosphatase (BALP), bone gla protein (BGP) and tartrate resistant acid phosphatase (TrACP) of two groups were evaluated. Results The BMD increased significantly after treatment in both groups (P<0.05), and the therapeutic efficacy of the treatment group was better than of the control group (P<0.05). After treatment, the level of BALP (308.76 ± 10.99 U/L vs. 280.00 ± 7.44 U/L, t=8.170) and the BGP (42.21 ± 3.04 ng/ml vs. 34.38 ± 2.06 ng/ml, t=6.818) of the treatment group were significantly higher than those of the control group (P<0.01). The level of TrACP decreased significantly after treatment in both groups (P<0.05), and the TrACP (60.12 ± 4.58 U/L vs. 67.25±4.06 U/L, t=1.653) of treatment group was significantly lower than that of the control group (P<0.05). Conclusions The Bushen-Qiangjin capsule can produce a content efficacy in treating AIBL in breast cancer patients, improving the BMD and bone metabolism.

Objective:The aim of this study was to develop and validate a new liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay for the quantification of melatonin in human serum.Methods:We describe the performance and validation of melatonin by LC-MS/MS. 182 serum samples from the patients diagnosed with Sleep disturbance who visited the Department of Psychiatry at Zhongshan Hospital affiliated to Fudan University from February 2022 to March 2023(56 males,162 females,mean age [45.51±16.31]years), as well as 182 healthy individuals were included(87 males,95 females,mean age [48.55±11.93]years). The two groups were used to assess the application of serum melatonin levels as a diagnostic indicator for sleep disorders (SDs). The liquid chromatography mass spectrometry (LC-MS) system with an chromatography column (2.1×100 mm, 1.8 μm) was used for separation. The column temperature was set at 35 ℃, as well as the mobile phase consisting of a 0.1% formic acid aqueous solution and pure acetonitrile. The flowing rate was set at 0.4 ml/min for gradient elution. The LC-MS/MS method was validated according to guidance documents, including the following parameters: specificity, selectivity, matrix effect, carryover contamination and reproducibility, lower limit of measuring interval, linearity, precision, recovery rate, dilution consistency, and serum sample stability. Then, it was subsequently employed to profile melatonin changes in Sleep disturbance.Results:The lower limit of quantification for melatonin was 1 pg/ml, and the linear range of detection was 1 pg/ml to 500 pg/ml ( r=0.999). The intra-day and intra-batch precision, expressed as the coefficient of variation ( CV), was within the range from 3.07% to 6.86%, which met the requirement of less than 15%. The recovery rate of the spiked samples ranged from 105.91% to 116.30%. The level of serum melatonin in the sleep disturbance group was significantly lower than that in the healthy control group ([2.00(1.00,3.28)] vs [8.35(4.28,14.80)] pg/ml, P<0.001). Conclusions:The LC-MS/MS method we developed for the quantification of melatonin is clinical practicable.

Objective:To investigate the perioperative impact of closure of mesenteric fissure in laparoscopic right hemicolectomy.Methods:The prospective randomized controlled trial was conducted. The clinical data of 320 patients who underwent laparoscopic right hemicolectomy in 11 medical centers, including The First Affiliated Hospital of China Medical University et al, from November 2022 to August 2023 were selected. Based on block randomization, patients were alloca-ted into the mesenteric fissure non-closure group and the mesenteric fissure closure group. Observa-tion indicators: (1) grouping of the enrolled patients; (2) intraoperative conditions; (3) postopera-tive conditions. Measurement data with skewed distribution were represented as M( Q1, Q3) and com-parison between groups was conducted using the Mann-Whitney U test. Count data were represen-ted as absolute numbers or percentages, and comparison between groups was conducted using the chi-square test or Fisher's exact probability. Comparison of ordinal data was conducted using the rank sum test. Comparison of visual analog scores was analyzed using generalized estimating equations. Results:(1) Grouping of the enrolled patients. A total of 320 patients with colon cancer were screened for eligibility, including 156 males and 164 females, aged 68(59,73)years. All the 320 patients were allocated into the mesenteric fissure non-closure group with 164 cases and the mesenteric fissure closure group with 156 cases. There was no significant difference in the age, body mass index, American Society of Anesthesiologist score, maximum tumor diameter, anastomosis location, anastomosis method, surgical approach, range of lymph node dissection, tumor staging between the two groups ( P>0.05) and there was a significant difference in the sex between them ( P<0.05). (2) Intraoperative conditions. There was no significant difference between the mesenteric fissure closure group and the mesenteric fissure non-closure group in the volume of intraoperative blood loss, operation time, conversion to laparotomy, intraoperative complication ( P>0.05). Three patients in the mesenteric fissure non-closure group were converted to laparotomy. One patient in the mesenteric fissure closure group was converted to laparotomy, and 2 cases with intraoperative complication were mesenteric hematoma. (3) Postoperative conditions. There was no significant difference between the mesenteric fissure non-closure group and the mesenteric fissure closure group in the overall postoperative complications ( χ2=0.28, P>0.05). There was no significant difference in the occurrence of postoperative intestinal obstruction, abdominal distension, ascites, pleural effusion, gastric paralysis, anastomotic bleeding, anastomotic leakage, or surgical wound infection between the two groups ( P>0.05). There was no significant difference between the two groups in the reoperation, postoperative gastric tube replacement. There was no significant differ-ence in time to postoperative first flatus, time to postoperative initial liquid food intake, time to post-operative resumption of bowel movements, duration of postoperative hospital stay, total hospital expenses between the two groups ( Z=-0.01, 0.43, 1.04, -0.54, -0.36, P>0.05). One patient in the mesenteric fissure non-closure group received reoperation. No perioperative internal hernia or death occurred in either group. The visual analog score decreased with time in both groups. There was no significant difference in the visual analog score between the mesenteric fissure closure group and the mesenteric fissure non-closure group [ β=-0.20(-0.53,0.13), P>0.05]. Conclusion:Compared with closure of mesenteric fissure, non-closure of mesenteric fissure during laparoscopic right hemi-colectomy dose not increase perioperative complications or postoperative management risk.