ObjectiveTo investigate the optimal termination time for acupuncture in treating patients with acute peripheral facial paralysis and its cost-effectiveness. MethodsA total of 120 eligible patients with acute-stage peri-pheral facial paralysis were randomly assigned to either the mild dysfunction termination group and the complete recovery termination group， with 60 patients in each group. Both groups received the standard acupuncture treatment protocol. Treatment in the mild dysfunction termination group was terminated when the Sunnybrook facial grade scale （SFGS） score first reached or exceeded 83 points， while that in the complete recovery termination group was terminated when the SFGS score first reached or exceeded 95 points. Assessments were conducted before treatment， 6 and 12 months after onset. SFGS， facial disability index （FDI） including physical function （FDIp） and social function （FDIs）， self-rating anxiety scale （SAS）， and self-rating depression scale （SDS） scores were assessed before treatment， and 6 and 12 months after onset. Any acupuncture-related adverse events during treatment were recorded for safety evaluation. Treatment sessions and medical costs including direct costs， indirect costs， insurance coverage， total societal costs， and patient out-of-pocket expenses were also recorded， and an economic evaluation was conducted including cost-effectiveness ratio （CER） and incremental cost-effectiveness ratio （ICER）. ResultsUltimately， 56 patients in the mild dysfunction termination group and 55 in the complete recovery termination group completed the follow-up. At 6 and 12 months after onset， SFGS and FDIp scores in both groups improved significantly while FDIs， SAS and SDS scores decreased （P＜0.05）. Comparison of scores between groups 6 months and 12 months after onset showed no statistically significant differences （P＞0.05）. During the trial， the incidence of adverse events was 13.3% （8/60） in the mild dysfunction termination group and 18.3% （11/60） in the complete recovery termination group， with no statistically significant difference （P＞0.05）. The number of treatment sessions， total social costs， and out-of-pocket expenses in the mild dysfunction termination group were significantly lower than those in the complete recovery termination group （P＜0.05）. The CER of the mild dysfunction termination group in SFGS， FDIp， FDIs， SAS， and SDS scores was lower than that of the complete recovery termination group. The ICER analysis showed that continuing treatment until full recovery incurred an additional cost of 573.30 CNY/point in SFGS improvement， whereas 1-point improvement in FDIp， FDIs， SAS， and SDS required 21,355.25 CNY， 1779.60 CNY， 3713.96 CNY， and 2755.52 CNY， respectively. ConclusionFor acupuncture in treating acute peripheral facial palsy， terminating treatment when mild dysfunction is achieved yields long-term efficacy comparable to that of continuing treatment until complete recovery， while significantly reducing medical costs and socioeconomic burden.

Objective:To investigate the effect of estrogen-related receptor α (ESRRA)-mediated lipophagy on the proliferation and migration abilities of nasopharyngeal carcinoma cells.Methods:A total of 16 clinical samples diagnosed by pathology in the Affiliated Hospital of Nantong University from 2021 to 2023 were selected, including 8 normal nasopharyngeal mucosa tissues and 8 nasopharyngeal carcinoma tissues. Immortalized normal nasopharyngeal epithelial cell line NP69 and nasopharyngeal carcinoma cell lines C666-1, CNE2, TW03-EBV and TW03 were selected. The cell lines C666-1 and CNE2 were divided into the siR-NC group (transfected with small interfering RNA negative control sequence) and siR-ESRRA group (transfected with small interfering RNA against ESRRA gene). The relative expression levels of ESRRA were detected by Western blotting and immunohistochemical assay. EdU assay was used to detect the proliferation ability of C666-1 and CNE2 cells, and Transwell assay was used to detect the migration ability. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression levels of ESRRA and perilipin 3 (PLIN3) mRNA. The formation of lipophagy in C666-1 and CNE2 cells was observed by transmission electron microscopy. The co-localization of LC3, PLIN3 and LAMP2 with lipid droplets labeling with Bodipy was detected by immunofluorescence assay. Dual-luciferase reporter gene assay was used to verify the targeting relationship between ESRRA and PLIN3.Results:The relative expression level of ESRRA in nasopharyngeal carcinoma tissues was higher than that in normal nasopharyngeal mucosa tissues(1.15±0.75 vs. 0.32±0.21, t = 3.02, P = 0.009). The relative expression level of ESRRA in nasopharyngeal carcinoma cell lines C666-1 (1.539±0.044), CNE2 (1.420±0.030), TW03-EBV (2.867±0.044), and TW03 (1.323±0.022) were higher than that in normal nasopharyngeal epithelial cell line NP69 (0.094±0.002), and the difference was statistically significant ( F = 34.08, P < 0.001).The results of EdU assay showed that the proportions of EdU labeled positive cells in CNE2 cells of siR-NC group and siR-ESRRA group were (70.44±4.06)% and (51.51±0.92)% ( t = 7.88, P = 0.001), and the proportions in C666-1 cells were (62.25±3.89)% and (54.91±0.27)% ( t = 3.26, P = 0.031). The results of Transwell assay showed that the number of migrating cells in CNE2 and C666-1 cells was less than that in siR-NC group [CNE2 cells: (181±7) cells vs. (261±21) cells; C666-1 cells: (201±16) cells vs. (256±7) cells], and the differences were statistically significant ( t = 6.30, P = 0.003; t = 5.43, P = 0.006). According to qRT-PCR results, the relative expression level of PLIN3 mRNA in the siR-ESRRA group was higher than that in the siR-NC group (CNE2 cells: 1.58±0.16 vs. 0.83±0.17, t = 5.59, P = 0.005; C666-1 cells: 1.37±0.12 vs. 1.06±0.06, t = 3.86, P = 0.018). The dual-luciferase reporter gene assay results indicated a targeted binding interaction between PLIN3 and ESRRA. Transmission electron microscopy observation showed that the lipid droplets in nasopharyngeal carcinoma cells increased and the binding to autophagosomes decreased after knockdown of ESRRA. The results of immunofluorescence assay demonstrated that, in contrast to the siR-NC group, there was a decrease in the co-localization of LC3 and LAMP2 and an increase in the co-localization of lipid droplets with PLIN3. Conclusions:ESRRA is highly expressed in nasopharyngeal carcinoma tissues and cells. As a transcription repressor, ESRRA may work to prevent PLIN3 from being transcribed, decrease lipid droplet stability, mediate lipophagy, and promote proliferation and migration of nasopharyngeal carcinoma cells.

Background: Colorectal cancer is one of the most commonly diagnosed cancer and leading causes of cancer death worldwide. Fibrinogen-like protein 2 (FGL2), a member of the fibrinogen family, has been demonstrated as a regulator of immune cell functions in tumor microenvironment and might facilitate tumor progression. Aims: To study the expression and clinical significance of FGL2 in colon cancer. Methods: One hundred and fifty colon cancer patients diagnosed from January 2018 to January 2022 at the Second Hospital of Tianjin Medical University were enrolled in this study. The paraffin-embedded tissues were collected for detection of FGL2 protein and the surface markers of tumor infiltrating immune cells (TIICs), including CD4, CD8, CD20, CD68, and CD56 by using immunohistochemistry. The correlations of FGL2 expression level with the clinicopathological parameters and TIICs counting were analyzed. Results: Expression of FGL2 was upregulated in 68.7% of the colon cancer cases. Its expression level was correlated positively with the tumor size and TNM staging (all P<0.05), while no correlations were found between FGL2 expression level and gender, age, tumor differentiation, and presence of vascular invasion (all P>0.05). Meanwhile, the expression level of FGL2 was associated with the cell counting of CD4

ObjectiveTo investigate the changes and potential effects of differentially expressed microRNAs (miRNAs) in the development and progression of liver injury in a mouse model of autoimmune hepatitis (AIH) induced by concanavalin A (ConA). MethodsEight healthy male specific pathogen-free C57BL/6 mice were randomly divided into model group and control group, with four mice in each group. The mice in the model group were given tail vein injection of ConA 15 mg/kg, and those in the control group were given an equal volume of normal saline. All mice were sacrificed after 8 hours of modeling, Total RNA in liver tissue was extracted, gene microarray was used to screen out differentially expressed miRNAs, and target prediction and function analysis were performed for upregulated and downregulated miRNAs. The independent samples t-test was used for comparison of differentially expressed miRNAs between two groups. ResultsThe principal component analysis showed that the inter-group difference of the data extracted by gene microarray met the conditions for further analysis. Compared with the control group, the model group had 31 upregulated miRNAs and 18 downregulated miRNAs in mouse liver, which had a regulatory relationship with 959 target genes (601 upregulated genes and 358 downregulated genes). GO analysis showed that in the model group, the target genes of the upregulated miRNAs mainly had the molecular functions such as “DNA binding” (P=1.47×10-6), participated in the biological processes such as “transcription, DNA-templated” (P=2.36×10-7), and were mainly enriched in the cellular components such as “neuronal cell body” (P=5.99×10-6), while the target genes of the downregulated miRNAs had the molecular functions such as “RNA polymerase II proximal promoter sequence-specific DNA binding” (P=2.49×10-6), participated in the biological processes such as “regulation of transcription, DNA-templated” (P=1.64×10-11), and were mainly enriched in the cellular components such as “nucleoplasm” (P=4.30×10-10). KEGG pathway enrichment analysis showed that the target genes of the upregulated miRNAs were mainly enriched in “Endocytosis” (P=0.000 4), while the target genes of the downregulated miRNAs were mainly enriched in the “Hippo signaling pathway” (P=0.004), and the above functional analysis results were statistically significant (P＜0.05). ConclusionThere are differentially expressed miRNAs in the pathogenesis of AIH, and these differentially expressed miRNAs can provide new targets for the clinical treatment of AIH.

Objective To explore the security of exosome derived from human umbilical cord mesenchymal stem cell(huc-MSC-exosome)as a kind of biological product on clinical application.Methods huc-MSC-exosome were separated by ultea centrifuge. CD9 and CD63 expression of huc-MSC-exosome were tested by Western-blot.Nanoparticle analysis was used to detect the size and concentration of huc-MSC-exosome.Hemolytic experiment,systemic anaphylaxis and blood routine test were used to verify the self-safety of huc-MSC-exosome.Results huc-MSC-exosome expressed specific markers of exosome,CD9 and CD63.Moreover,huc-MSC-exosome cannot result in hemolysis,systemic anaphylaxis reaction and abnormality of blood routine test.Conclusion Exosome derived from human umbilical cord mesenchymal stem cell has general signs of exosomes and possesses the certainly security,which may provide the experimental evidence for huc-MSC-exosome clinical application.

Objective The study was designed to analyze the efficacy of standardized house dust mite allergen vaccine to allergic rhinitis by subcutaneous injection and investigate the possible mechanism of specific immunotherapy(SIT).Methods From January 2011 to December 2011 a prospective study was performed in the Department of Otorhinolaryngology Head and Neck Surgery,the Affiliated Hospital of Nantong University,involving 90 patients with perennial AR,of whom 60 patients received Der p-SIT +pharmacotherapy after their approval and 30 received only pharmacotherapy.All patients were allergic to house dust mites.Symptom and medication scores were recorded three times:before the treatment,at the middle of treatment and at the end of treatment.Over a period of 1 yr.prior to and at the end of treatment,CD4 + CD25 + Foxp3 + Treg cells and Th17 cells were measured by flow cytometry.SPSS 21.0 software was used to analyze the data.Results The symptom scores using VAS and medication scores in AR patients treated with SIT and medication were reduced,the differences were significant (14.25 ± 6.40,1.00 ± 0.84 vs.32.18 ± 7.78,3.12 ± 1.54,t value was 19.65,10.71,both P ＜ 0.05).The symptom of VAS score in medication group was reduced after treatment (30.30 ±5.97 vs.20.30 ±5.79,t =10.09,P ＜0.05),but the medication score had not significant difference (P ＞ 0.05).The frequency of Thl7 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT,whereas the frequency of Treg cells were increased (x2 value was 2.81,2.80,both P ＜ 0.05),but not in medication group.Conclusions Both SIT and pharmacotherapy can improve symptoms of allergic rhinitis,but SIT can also reduce medication use.The effect of immunotherapy is better than drug treatment alone.The frequency of blood Th17 cells in peripheral blood mononuclear cells were decreased in patients treated with SIT,whereas the frequency of Treg cells were increased.

OBJECTIVETo study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat.

METHODSThirty healthy SD rats were divided into control group (n = 6) and injury group (n = 24) according to the random number table. Two round full-thickness skin defects (11 mm diameter) were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were disinfected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day (PID) 1, 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area (wound healing rate was calculated). Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 (the surface marker of macrophage) in the wound tissue was observed with immunohistochemical staining. The double positive expressions of induced nitric oxide synthase (iNOS) plus CD68 (type I macrophage) and arginase 1 (Arg-1) plus CD68 (type II macrophage) were observed with immunofluorescence staining. The levels of interferon-γ (IFN-γ), TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tissues (11 mm diameter) in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analysis of variance or LSD- t test.

RESULTSWound area of rats in injury group was gradually reduced after injury, and the overall difference of the wound healing rate on each PID was statistically significant (F = 358.55, P < 0.01). No abnormal appearance of skin tissue was observed in rats of control group. In injury group, inflammatory cell infiltration was obvious in wound tissue on PID 1 and 3; vascular structure and fresh collagen were observed in wound tissue on PID 7 and 13. Numbers of CD68 positive cells in skin tissue of rats in control group and wound tissue of rats in injury group on PID 1, 3, 7, and 13 were respectively (2.7 ± 1.5), (31.8 ± 3.5), (40.8 ± 4.7), (20.8 ± 2.8), (3.2 ± 2.4) per 200 times visual field (F = 180.55, P < 0.01). Compared with that in control group, the number of CD68 positive cells of rats in injury group was increased on PID 1, 3, and 7 (with t values respectively 18.81, 18.79, 14.05, P values below 0.01). No double positive expression of iNOS plus CD68 or Arg-1 plus CD68 was observed in normal tissue of rats in control group. In injury group, proportions of iNOS plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively (12.2 ± 2.8)%, (16.5 ± 2.9)%, (4.2 ± 2.3)%, (0.7 ± 0.8)% (F = 72.50, P < 0.01); proportions of Arg-1 plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively 0, (8.2 ± 1.9)%, (21.5 ± 3.4)%, (4.7 ± 2.0)% (F = 120.93, P < 0.01). In injury group, proportion of iNOS plus CD68 double positive cells on PID 3 was significantly higher than that on other PID (with t values respectively 2.65, 8.17, 12.95, P values below 0.05); proportion of Arg-1 plus CD68 double positive cells on PID 7 was higher than that on other PID (with t values respectively 15.27, 8.25, 10.38, P values below 0.01). Compared with that of Arg-1 plus CD68 double positive cells, proportion of iNOS plus CD68 double positive cells was higher on PID 1 and 3 (with t values respectively 10.71 and 5.88, P values below 0.01) and lower on PID 7 and 13 (with t values respectively 10.24 and 4.60, P values below 0.01). The overall differences of IFN-γ, TNF-α, IL-4, IL-13, and IL-10/IL-12 ratio in skin tissue of rats in control group and wound tissue of rats in injury group on every PID were statistically significant (with F values from 14.08 to 631.03, P values below 0.01). Compared with those in control group, levels of IFN-γ, TNF-α, IL-4, and IL-13 in wound tissue of rats in injury group were significantly higher on every PID (with t values from 4.58 to 9.17, P values below 0.05), while IL-10/IL-12 ratio was significantly higher on PID 1, 3, and 7 (with t values respectively 27.70, 30.51, 9.49, P values below 0.05) . In injury group, IFN-γ level on PID 1 [(61 ± 5) pg/mL] and IL-10/IL-12 ratio on PID 3 (1.647 ± 0.098) were significantly higher than those of control group and those on other PID in injury group [with IFN-γ level respectively (32 ± 4), (54 ± 6), (46 ± 7), (47 ± 4) pg/mL and IL-10/IL-12 ratio respectively 0.328 ± 0.045, 0.960 ± 0.034, 0.530 ± 0.028, 0.289 ± 0.040, with t values respectively from 3.19 to 8.20 and from 16.59 to 31.84, P values below 0.05].

CONCLUSIONSMacrophage infiltration increases in the healing process of full-thickness wound in rat with different phenotypes, among which type I macrophage appears in the inflammatory stage, and type II macrophage predominates in the proliferative stage.

Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, Myelomonocytic ; genetics ; metabolism ; Collagen ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukin-13 ; Interleukin-4 ; Macrophages ; Male ; Phenotype ; Rats ; Skin ; injuries ; Tumor Necrosis Factor-alpha ; blood ; Wound Healing ; genetics

Objective To explore the health-relate quality of life(QOL) status of adult persistent allergic rhinitis (PAR);the change of QOL of pro-post specific immunotherapy (SIT) and pharmacotherapy .Methods Skin prick tests(SPT) were performed on PAR patients .According to the results ,80 adult cases that were allergic to dermatophagoides were enrolled in ENT outpatient clinic of affiliated hospital of Nantong University from April to August 2011 .The patients were randomly allocated to receive either specific immunotherapy(n=40) or pharmacotherapy (n=40) ,all of them were given RQLQ before and after half-year treatment ;40 cases without any allergic diseases were chosen from ENT in-patient department ,and were given RQLQ .The scores of previous treatment of the PAR group were compared with health control group ,then compared with the scores of post-treatment ,and also compared the scores of post-treatment between the immunotherapy group and pharmacotherapy group .Results The scores of the PAR patients were higher than that of health control patients in all dimensions of RQLQ (P< 0 .05) ,and the most troublesome problems were nasal symptoms .The scores of the patients who received SIT were evidently lower than that of pro-treatment in all dimensions of RQLQ(P<0 .05) ,the scores of the patients who received medical treatment were also lower than before (P<0 .05) , and the scores of the SIT group were lower than the pharmacotherapy group (P<0 .05) .Conclusion The QOL of adult patients with PAR was improved after SIT or drug treatment ,and QOL improvement is more obvious treat by SIT .

Objective To compare the efficacy and safety of tranexamic acid(TA)and norethisterone(NET)for the treatment of patients with ovulatory menorrhagia in China. Methods Onehundred and thirty one patients with proven ovulatory menorrhagia from gynecologic clinics of 5 teaching hospitals located in 4 different cities in China were enrolled during Jul 2004 to Dec 2006.Ameng them 128 completed the study.Patients were randomly divided into two therapeutic regimen groups:TA 1g thrice daily during menstrual cycle days(D)1-5,69 cases;or NET 5 mg twice daily on D19-26.59 cases.The drugs were administered for 2 consecutive cycles,then withdrawn and patients were followed-up for 1 more cycle.Data on menstrual blood loss [ estimated by pictorial blood assessment chart(PBAC)],length of menstrual periods,quality of life(QOL)evaluated by a 6 item health-related questionnaire were collectedbefore,during each cycle and were compared.Results Both treatments led to significant decreases of mean PBAC scores and shorter duration of menstrual periods,and improved the QOL ranking during the twotreatment cycles.The mean percentages of PBAC decrements in the TA first and second cycles were significantly greater than those in the NET corresponding cycles(35%VS 17%,P=0.004;4J4%VS 34%,P=0.04 respectively).The success rate of TA second cycle was higher than that of the NET second cycle (41%VS 24%,P=0.04).Improvement of QOL ranking in the TA first cycle was also significantly better than those in the NET first cycle ( P=0.03).The percentage of patients with at least 1 adverse event in TA group(19%)was significantly lower than that in NET group(35%,P=0.04).Patients'willingness tocontinue the treatment in the TA second and follow-up cycles(94%,79%respectively)were significantly higher than those in the corresponding cycles of NET groups(79%,59%respectively;P=0.01,P=0.02).Conclusion The regimen of TA 3 g daily during menstrual days 1-5 is a more effective and tolerable treatment than luteal phase norethisterone for patients with ovulatory menorrhagia.