Objective:To evaluate the feasibility of indocyanine green fluorescence imaging technology applied in thyroid surgery to identify parathyroid gland.Methods:From Oct 2021 to May 2022, data of 42 patients undergoing thyroidectomy via gasless unilateral axillary approach in Zhejiang Provincial Hospital of Traditional Chinese Medicine were retrospectively analyzed. Cases using intraoperative fluorescence imaging technology(42 cases) were compared with conventional laparocopic approach.Results:The number of parathyroid glands dissected in the study group 1.57±0.61 was higher than that in the control group 0.56± 0.59 ( t=-5.472, P<0.05). The PTH value of the study group was (2.88±1.23)pmol/L on the first day after operation, which was higher than that of the control group (2.16±0.10)pmol/L ( t=-1.844, P<0.05). The blood parathyroid hormone value on the third day(3.22±1.31)pmol/L was higher than that of the control group (2.55±0.81) pmol/L ( t=-2.041, P<0.05). There were 2 cases of hypoparathyroidism in the study group, less than 5 cases in the control group, but there was no significant difference between the two groups( χ2=0.942, P>0.05). There was 1 case of hypocalcemia in the study group and 3 cases in the control group ( χ2=0.731, P>0.05). Conclusion:Using indocyanine green fluorescence imaging technology to identify parathyroid gland is feasible, simple, fast, safe and effective.

Objective:To observe and analyze the effect of peripore cavity size on visual function of macular area before and after surgery for idiopathic macular hole (IMH).Methods:A retrospective clinical study. From July 2020 to February 2021, a total of 25 patients with 25 eyes with monocular IMH (operation group) diagnosed by ophthalmology examination in Department of ophthalmology, Fourth Hospital of Hebei Medical University were included in the study. The control group was contralateral healthy eyes. All subjects were examined by best corrected visual acuity (BCVA), microfield of vision, frequency domain optical coherence tomography (SD-OCT), and OCT angiography (OCTA). The diameter of macular hole was measured by SD-OCT. The cystic morphology of deep capillary plexus (DCP) was detected by en face OCT, and the cystic area was measured by Image J software. MP-3 microperimeter was used to measure central macular retinal light sensitivity (MS) and mean macular retinal light sensitivity (MMS). Central macular retinal light sensitivity (CMS), MMS and cystic cavity MS were measured in the operation group. MMS was measured in the control group. The microperimetry images were superimposed on the DCP layer of OCTA to identify and calculate the average MS within the lumen and compare it with the control group. Standard three incisions were performed in all affected eyes by vitrectomy of the flat part of the ciliary body + stripping of the inner boundary membrane + intraocular sterile air filling. Three months after the operation, the same equipment and methods were used to perform relevant examinations. Paired sample t test was used to compare MS between operation group and control group. Pearson correlation analysis was used to analyze the correlation between capsular area, macular hole diameter before and after operation and MS before and after operation. The correlation between BCVA and capsular area before and after surgery was analyzed by Spearman correlation analysis. Results:In the surgical group, the retinal MS was (4.24±3.07) dB. The MMS of control group was (19.08±6.11) dB. The MS in the surgical group was significantly lower than that in the control group, and the difference was statistically significant ( t=10.832, P<0.01). Before operation, the area of cyst was (1.04±0.55) mm 2, and the diameter of macular hole was (564.80±166.59) μm. CMS and MMS were (2.27±2.29) dB and (9.08±3.65) dB, respectively. The diameter of macular hole ( r=0.50, P=0.010) and BCVA before operation ( r=0.57, P<0.001) were positively correlated with peripore cavity area. Before operation, CMS and MMS were negatively correlated with peripore cavity area ( r=-0.53, -0.47; P=0.010, 0.020). At 3 months after surgery, the capsular area was negatively correlated with CMS and MMS ( r=-0.65,-0.76; P=0.020, 0.030). There was no correlation with BCVA ( r=0.23, P=0.470). Conclusions:Retinal MS is decreased in the peri-capsular area of IMH pore. There is a positive correlation between capsule area, BCVA and macular hole diameter before operation. The capsular area is negatively correlated with CMS and MMS before operation.

Objective:To investigate the association between relative mitochondrial DNA copy number (mtDNA-CN) and coal-burning-borne endemic fluorosis (abbreviated as coal-burning-borne fluorosis).Methods:From June 2018 to March 2019, using cross-sectional study, 482 patients with coal-burning-borne fluorosis were selected as the case group in Bijie City, a typical coal-burning-borne fluorosis area of Guizhou Province; meanwhile, 212 healthy individuals from Changshun County, a non-coal-burning-borne fluorosis area in Guizhou Province, were selected as the control group. Questionnaire survey and physical examination were used to collect general condition such as basic information and living habits of the two groups, peripheral venous blood samples were collected, and real-time fluorescence quantitative PCR was used to detect the relative mtDNA-CN in peripheral blood. The correlation between relative mtDNA-CN and coal-burning-borne fluorosis was analyzed by binary and unordered multi-class logistic regression.Results:There were significant differences in the body mass index (BMI), and the distribution of gender rario, marital status and education level between the control group and the case group ( t = 7.91, χ 2 = 5.11, 13.33, 34.32, P < 0.05). The relative mtDNA-CN in the control group was higher than that in the case group [median (quartile): 202 (138, 292) vs 131 (96, 217), Z = - 7.80, P < 0.001]. The results of binary logistic regression analysis [odds ratio (95% confidence interval)] showed that educational level [primary school: 0.572 (0.377 - 0.868), junior high school and above: 0.292 (0.174 - 0.493)], relative mtDNA-CN [131 - < 217: 0.265 (0.144 - 0.488), ≥217: 0.183 (0.100 - 0.335)] and BMI [1.222 (1.142 - 1.307)] were the influencing factors for the risk of coal-burning-borne fluorosis( P < 0.05). In subgroups with different BMI and educational levels, the relative mtDNA-CN was significantly negatively correlated with the risk of coal-burning-borne fluorosis( Ptrend < 0.05), and there was no interaction between mtDNA-CN and BMI and educational levels ( Pinteraction > 0.05). The results of unordered multi-class logistic regression analysis showed that the relative mtDNA-CN were significantly negatively correlated with the risk of dental fluorosis and skeletal fluorosis ( Ptrend < 0.05). Conclusion:The higher the relative mtDNA-CN, the lower the risk of coal-burning-borne fluorosis, suggesting that mtDNA-CN may be a potential biomarker of coal-burning fluorosis.

Objective:To explore the relationship between children's intelligence and urinary fluoride in Suojia Township of Miao, Yi and Hui Nationalities (referred to as Suojia Township), a coal-burning-borne endemic fluorosis area in Guizhou Province.Methods:In April 2019, 173 children aged 10 to 13 years old were selected from three schools in Suojia Township. According to whether they had dental fluorosis, the children were divided into case group ( n = 104) and control group ( n = 69). Middle segment urine samples of the children were collected and urinary fluoride level was determined by the method of ion-selective electrode. Combined Raven's Test-the Rural in China (CRT-RC2) was used for children's intelligence quotient (IQ) test. Linear regression analysis was used to observe the association between urinary fluoride and IQ, and the results were expressed by regression coefficient ( β) and 95% confidence interval ( CI). Results:Urinary fluoride level of case group was higher than that of control group [(2.14 ± 1.78) vs (1.53 ± 0.98) mg/L], and IQ was lower than that of control group [(92.33 ± 11.68) vs (100.38 ± 11.87) points], and the differences were statistically significant ( t = 2.58, 4.41, P < 0.05). The linear regression equation of urinary fluoride ( X) and IQ ( Y) of case group was Y = 96.99 - 2.86 X. For every 1 mg/L increase in urinary fluoride level, IQ decreased by 2.86 points ( β = - 2.86, 95% CI: - 5.48 - - 0.24). Conclusion:Long-term exposure to fluoride pollution from coal burning may damage children's intelligence, and children's IQ decreases with increase of fluoride level in urine.

Bisecting N-acetylglucosamine(GlcNAc),a GlcNAc linked to the core β-mannose resi-due via a β1,4 linkage,is a special type of N-glycosylation that has been reported to be involved in various biological processes,such as cell adhesion and fetal development.This N-glycan structure is abundant in human trophoblasts,which is postulated to be resistant to natural killer cell-mediated cytotoxicity,enabling a mother to nourish a fetus without rejection.In this study,we hypothesized that the human amniotic membrane,which serves as the last barrier for the fetus,may also express bisected-type glycans.To test this hypothesis,glycomic analysis of the human amniotic membrane was performed,and bisected N-glycans were detected.Furthermore,our pro-teomic data,which have been previously employed to explore human missing proteins,were ana-lyzed and the presence of bisecting GlcNAc-modified peptides was confirmed.A total of 41 glycoproteins with 43 glycopeptides were found to possess a bisecting GlcNAc,and 25 of these gly-coproteins were reported to exhibit this type of modification for the first time.These results provide insights into the potential roles of bisecting GlcNAc modification in the human amniotic membrane,and can be beneficial to functional studies on glycoproteins with bisecting GlcNAc modifications and functional studies on immune suppression in human placenta.

Objective:To investigate the relationship between gene polymorphism of osteoprotegerin (OPG) and coal-burning endemic fluorosis in Guizhou Province.Methods:In 2018 and 2019, a case-control study was conducted in Bijie City, a typical coal-burning endemic fluorosis area in Guizhou Province, and 260 cases of coal-burning endemic fluorosis patients were selected as case group. According to the "Diagnostic Criteria for Endemic Skeletal Fluorosis" (WS 192-2008), the case group was divided into severe group (130 cases) and mild group (130 cases). At the same time, 130 cases without dental fluorosis and skeletal fluorosis symptoms were chosen as control group in Changshun County, a non-coal-burning endemic fluorosis area in Guizhou Province. Whole blood genomic DNA was extracted, and real-time fluorescence quantitative PCR with TaqMan-MGB probe was utilized to type the OPG gene rs2460985, rs2073618, rs6469804 and rs6993813 four single nucleotide polymorphism (SNP) loci of all samples, and genetic model analysis was performed to compare the frequency distribution of its alleles, genotypes and constructed haplotypes in control, mild and severe groups. Results:By Hardy-Weinberg equilibrium test, the genotype frequencies of the four SNP loci all reached genetic balance in control, mild and severe groups ( P > 0.05). There was a statistically significant difference in the genotype frequency of OPG gene rs6469804 locus among the three groups (χ 2 = 10.615, P < 0.05), and the difference in the genotype frequency of this locus between the control group and the severe group was statistically significant (χ 2 = 6.784, P < 0.05). The results of genetic model analysis showed that in comparison between the control group and the severe group, the optimal genetic model of rs6469804 locus was the overdominant genetic model, the frequency distribution of genotypes AA + GG and AG in the control group and the severe group was statistically significant [odds ratio ( OR) = 1.94, 95% confidence interval ( CI): 1.16 - 3.23, P < 0.05], genotype AG was a risk factor for coal-burning endemic fluorosis. In comparison between the control group and the mild group, the optimal genetic model of rs2073618 locus was the recessive genetic model, the frequency distribution of genotypes GG + GC and CC in the control group and the mild group was statistically significant ( OR = 3.17, 95% CI: 1.08 - 9.30, P < 0.05), genotype CC was a risk factor for coal-burning endemic fluorosis. In comparison between the control group and the mild group, haplotypes C-C-G-T and T-G-A-C were risk factors for coal-burning endemic fluorosis (adjusted OR = 2.41, 1.98, 95% CI: 1.29 - 4.50, 1.22 - 3.23, P < 0.05); in comparison between the control group and the severe group, haplotype T-G-A-C was a risk factor for coal-burning endemic fluorosis (adjusted OR = 1.87, 95% CI: 1.14 - 3.07, P < 0.05). Conclusion:OPG gene rs6469804 locus genotype AG and rs2073618 locus genotype CC may be risk factors for coal-burning endemic fluorosis.

OBJECTIVETo explore the nerve regeneration mechanism of electroacupuncture (EA) combined with polysaccharide of gastrodia elate blume (PGB) for secondary thalamic damage of focal cerebral ischemia.

METHODSForty Sprague-Dawley adult rats were randomly divided into a normal control group, a model group, an EA group, a PGB group and an EA + PGB group, 8 rats in each group. The rat model of right middle cerebral artery occlusion was prepared by suture-occluded method. Two weeks after model establishment, rats in the normal control group and model group received no treatment; rats in the EA group were treated with EA at "Baihui" (GV 20) and left "Zusanli" (ST 36), 30 min per treatment, once a day for 14 successive days; rats in the PGB group were treated with intragastric administration of PGB (100 mg/kg) , once a day for 14 days; rats in the EA + PGB group were treated with EA and PGB treatment, once a day for totally 14 days. The expressions of nestin and stem cell factor (SCF) in thalamic ventroposterolateral nucleus (VPL) were detected by immunohistochemical method.

RESULTSThere were positive cells of nestin in ischemia VPL in the model group, and the number of SCF positive cells was increased compared with that in the normal control group (P<0.05). The number of positive cells of nestin and SCF in ischemia VPL in the EA group, PGB group and the EA + PGB group was increased compared with that in the model group (all P<0.05), and the average gray value of immune positive product was all reduced (all P<0.05). The number of positive cells of nestin and SCF in the EA + PGB group was higher than that in the EA group or the PGB group (all P<0.05).

CONCLUSIONEA combined with PGB can significantly increase the SCF expression in ischemia VPL and promote the proliferation of neural stem cells, which is likely to be one of the nerve regeneration mechanism of acupuncture and medication tor secondary thalamic damage of local cerebral isctemia.

Animals ; Brain Ischemia ; drug therapy ; genetics ; metabolism ; therapy ; Combined Modality Therapy ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Electroacupuncture ; Gastrodia ; chemistry ; Humans ; Male ; Nestin ; genetics ; metabolism ; Polysaccharides ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Stem Cell Factor ; genetics ; metabolism ; Thalamic Nuclei ; metabolism

A global quality control method based on high performance liquid chromatography (HPLC) coupled with diode array detection (DAD), single quadrupole mass spectrometry (MS) and time-of-flight mass spectrometry (TOFMS) was developed for simultaneous determination of seven major components (mangiferin, neomangiferin, timosaponin E1, timosaponin E, timosaponin BⅡ, timosaponin BⅢ, and timosaponin AⅢ) and identification of most components in extracts of Rhizoma Anemarrhenae (RA). HPLC analysis was performed on an Agilent SB-C18 column (4.6 mm×150 mm, 5 μm) by gradient elution using acetonitrile and water-acetic acid(100∶0.05, v/v) as the mobile phase. Seven major components in RA were successfully separated. This quantitative method was fully validated in respect of the following performance criteria: linearity, precision, repeatability, stability, accuracy, limits of detection (LOD) and quantification (LOQ). A formula database of known compounds in RA was established, against which, most of the reported components in this herbal extract were identified effectively based on the extract masses acquired by TOFMS. This qualitative and quantitative method was successfully used to analyze the components in 10 batches of RA samples collected from different regions in China. This global quality control method, which consisted of HPLC-DAD-MS assay of seven major components and unambiguous identification of nineteen components, is suitable for routine quantification and comprehensive quality control of RA.

Objective To study whether Helicobacter pylori CagA protein can control gastrin gene expression and the detailed mechanism. Methods First, pcDNA3. 1ZEO (-)/cagA7 was transfected into gastric cancer cell lines AGS and SGC-7901 cells. At the same time, culturing the Helicobacter pylori NCTC11637 and infecting AGS and SGC-7901 cells with it. Next, in the infected and transfecled AGS and SGC-7901 cells, respectively adding the JAK2 signaling pathway inhibitor AG490 and the ERK signaling pathway inhibitor U0126 to inhibit the two signaling pathway. Untreated gastric cancer cells and empty vector transfected cells as the control. Using real-time fluorescence quantitative PCR to detect the levels of gastrin mRNA in transfected and infected cells. Results After AGS and SGC-7901 cells were transfected with pcDNA3. lZE0(-)/cagA7 and infected with NCTC11637, the results showed that the expression of gastrin mRNA increased significantly (P ＜ 0. 05) in transfected and infected cells as compared with the control group, but after adding the inhibitor AG490 and U0126 respectively, the expression of gastrin mRNA decreased significantly(P＜0.05). Conclution These results suggest that CagA may up-regulate the expression of the gastrin gene, and CagA is one of the important proteins in regulating gastrin gene expression. The ERK/MAPK and JAK/STAT signaling pathways may be involved in controlling of gastrin gene expression by CagA.

BACKGROUNDHuman cytomegalovirus (HCMV) infection is an important infectious agent that results in neonatal disease and congenital deformity. HCMV infection may affect in many organs. The different symptoms and tissue tropism of HCMV infection perhaps resulted from the genetic polymorphism of HCMV. HCMV UL144 open reading frames encode a homologue of the tumor necrosis factor receptor. It seems important to study the strain-specific variability of UL144 sequence in low-passage clinical isolates and to discuss if the variability related to the clinical HCMV infection.

METHODSHCMV-UL144 gene was amplified by PCR assay in 65 low-passage clinical isolates and urine from 7 healthy children who were HCMV-DNA positive by quantitative PCR. All the positive PCR products were analyzed by Heteroduplex mobility assay and single-stranded conformation polymorphism (HMA-SSCP) and 32 of them were sequenced.

RESULTSFifty-five isolates and 5 urine specimens were HCMV-UL144 positive by PCR. Sequencing and HMA-SSCP analysis showed that significant strain-specific variability was present in the UL144 ORFs. Comparing UL144 sequences and the corresponding symptoms showed that genotype 2 did not exist in megacolon isolates. And genotype 1 and 3 were the major types among microcephaly and jaundice isolates respectively.

CONCLUSIONHCMV-UL144 existed in almost all the low passage isolates. HMA-SSCP assay is an easy and effective method to detect the genetype of HCMV-UL144 sequence. The characteristic of sequences in different isolates showed that UL144 gene may play an important role in HCMV infection.

Cytomegalovirus ; classification ; genetics ; isolation & purification ; Cytomegalovirus Infections ; virology ; Genotype ; Humans ; Infant ; Infant, Newborn ; Membrane Glycoproteins ; genetics ; Molecular Sequence Data ; Phylogeny ; Polymerase Chain Reaction ; Polymorphism, Single-Stranded Conformational ; Sequence Analysis, DNA ; Viral Proteins ; genetics