Objective To investigate the mechanism by which Erdong Xiaoke Formula(EDXKF)regulates lacrimal gland autophagy in type 2 diabetes-induced dry eye rats through the peroxisome proliferator-activated receptor γ(PPARγ)/mammalian target of rapamycin(mTOR)signal pathway.Methods Healthy male SD rats were fed with high-sugar and high-fat chow and then injected intraperitoneally 30 mg·kg-1 streptozotocin to construct type 2 diabetes-induced dry eye rat models.Healthy male SD rats were selected as a blank group.Type 2 diabetes rats were randomly divided into a model group,a Traditional Chinese Medicine(TCM)group(given 11 g·kg-1 of EDXKF through gavage),an antagonist group(given 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection),and a TCM plus antagonist group(given 11 g·kg-1 EDXKF through gavage and 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection).Fasting blood glucose(FBG),corneal fluorescein(FL)staining,tear film break-up time(BUT)and phenol red cotton thread test(Prtt)were examined before modeling,after modeling,and after intervention.The weight of the lacrimal gland was compared among different groups after sampling.Hematoxylin and eosin(HE)staining was performed to analyze lacrimal gland hismorphol-ogy.The expression of microtubule-associated protein 1 light chain 3(LC3)and Sequestosome-1(p62)in the lacrimal gland was examined by immunofluorescence.Western blot was used to detect the expression of PPARγ,p62,LC3 Ⅱ,LC3 I,mTOR and p-mTOR in the lacrimal gland.Results FBG levels in the antagonist group were significantly increased after intervention,compared with those in the TCM group(P＜0.01).Ocular surface examination showed that compared with the model group,the TCM group had increased BUT and Prtt scores and decreased FL scores,the antagonist group had de-creased BUT and Prtt scores and increased FL scores(all P＜0.05).Compared with the antagonist group,the TCM plus antagonist group showed increased BUT and Prtt scores and decreased FL scores(all P＜0.05).The weight analysis of lac-rimal glands revealed that the lacrimal gland weight increased in the TCM group and decreased in the antagonist group,compared with that in the model group(all P＜0.01).The lacrimal gland weight in the TCM plus antagonist group was higher than that in the antagonist group(P＜0.01).HE staining of the lacrimal gland showed atrophy of glandular lobules,increased fusion and expansion of follicles,and dense distribution of nuclei in model and antagonist groups.These symp-toms were more obvious in the antagonist group.Compared with antagonist and TCM plus antagonist groups,the TCM group showed improved symptoms,with tightly arranged follicles,partial atrophy and fusion,and a small amount of expan-sion.Immunofluorescence staining for detecting the average fluorescence intensity of LC3 and p62 showed that compared with the model group,the TCM group had increased LC3 levels and decreased p62 levels,and the antagonist group had de-creased LC3 levels and increased p62 levels(all P＜0.01).Compared with those in the antagonist group,LC3 levels were increased and p62 levels were decreased in the TCM plus antagonist group(both P＜0.01).Western blot results showed that compared with the model group,the TCM group had increased PPARγ and LC3 levels and decreased p62 and p-mTOR/mTOR levels.PPARγ and LC3 levels were decreased and p62 and p-mTOR/mTOR levels were increased in the antagonist group,compared with those in the model group.The TCM plus antagonist group had higher PPARγ and LC3 levels and low-er p62 and p-mTOR/mTOR levels than the antagonist group(all P＜0.05).Conclusion EDXKF can regulate the PPARy/mTOR signal pathway to promote lacrimal gland autophagy and thus alleviate dry eyes in type 2 diabetes rats.

Objective This study reveals the potential mechanism of electroacupuncture in alleviating the inflamma-tory injury of sensory nerves and alleviating the abnormal sensation on the ocular surface in type 2 diabetes rats with dry eye.Methods Healthy male SD rats were induced to establish type 2 diabetes dry eye rat model.Model rats were ran-domly divided into model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group in the 12th week of the experiment.The blank group was established as well,and each group was intervened for 2 weeks.The corneal fluorescein sodium staining(FL),break-up time(BUT),phenol red thread test(PRT),corneal mechanical per-ception threshold(CTT)and blink frequency were measured in each group of rats.After sampling,the expression of neu-ronal injury markers,neuron specific enolase(NSE),neurofilament light chain protein Nf Light(NF-L),phosphorylated microtubule associated protein(Phospho-TAU),as well as inflammatory factors of cysteine protease-1(Caspase-1),inter-leukin-18(IL-18),and interleukin-1β(IL-1β),were observed in the trigeminal ganglion(TG)and spinal cord caudate nu-cleus(SpVc)of each group of rats.Results At the end of the 14th week,compared with the blank group,the FL of the model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group significantly increased,while PRT and BUT significantly decreased.Moreover,the CTT and blink frequency of the model group,sham-acupuncture group,and fluorometholone group significantly decreased(all P＜0.01).Compared with the model group,the electroacu-puncture group and the fluorometholone group showed a decrease in FL,as well as an increase in BUT,PRT,CTT,and blink frequency,with statistically significant differences(all P＜0.05).Compared with the electroacupuncture group,the sham-acupuncture group showed significantly higher FL along with significantly lower values in BUT,PRT,CTT,and blink frequency(all P＜0.05),while the fluorometholone group exhibited decreased FL,PRT,CTT,and blink frequency but in-creased BUT(all P＜0.05).Compared to the sham-acupuncture group,the fluorometholone group exhibited significantly lower FL,but higher BUT and PRT(all P＜0.01).In TG and Sp Vc,compared with the blank group,the expression of NSE,NF-L and Phospho-TAU increased in the model group and the sham-acupuncture group(all P＜0.05);compared with the model group,the expression of NSE,NF-L and Phospho-TAU significantly reduced in the electroacupuncture group(P＜0.01);compared with the electroacupuncture group,the expression of NSE,NF-L and Phospho-TAU increased in the electroacupuncture group and the expression of Phospho-TAU in the sham-acupuncture group also increased(P＜0.05);compared with the sham-acupuncture group,the expression of Phospho-TAU or NF-L significantly reduced in the fluoro-metholone group(all P＜0.05).The Western Blot results indicate that compared with the blank group,the protein expres-sion of Caspase-1,IL-1β,and IL-18 in TG and SpVc of the model group increased;moreover,the expression of Caspase-1 and IL-18 proteins in the sham-acupuncture group also increased(all P＜0.05).Compared with the model group,the ex-pression of Caspase-1,IL-1β,and IL-18 proteins in TG and SpVc reduced in the electroacupuncture group and the fluoro-metholone group,and the expression of IL-1 β proteins in TG and SpVc also reduced in the sham-acupuncture group(all P＜0.05).Compared with the electroacupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc increased in the sham-acupuncture group(all P＜0.05).Compared with the sham-acupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc reduced in the fluorometholone group(all P＜0.05).Conclusion Electroacupuncture can effectively alleviate the abnormal ocular surface sensation of type 2 diabetes dry eye rats by alleviating the neuroinflammatory injury in TG and SpVc.

Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1％),and CDKN2A homozygous deletion(36.8％)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.

Objective:To analyze the human immunodeficiency virus (HIV) screening status and the resulting residual risk (RR) among blood donors across 17 provincial blood centers in China.Methods:This study used a cross-sectional study. Data on HIV infection markers per 100 000 first-time donors (FD) and repeat donors (RD) from January 2015 to December 2024 were extracted from the National Blood Establishment Performance Comparison Information Management System. Questionnaires were used to collect each center′s HIV screening strategy, algorithm, serological test (ST) kit manufacturers, gray-zone setting for ST, and nucleic acid test (NAT) modality, method, and platform. The incidence-window-period model was used to calculate the residual risk for first-time donors (RR FD), repeat donors (RR RD), and total donors (RR TD) at each center. Horizontal and vertical analysis of RR FD, RR RD, and RR TD across centers and years were performed. Results:All 17 centers applied the same HIV screening strategy which was two rounds of ST followed by one round of NAT. Eight of them operated a single screening algorithm, six employed two algorithms and three used three. Eleven centers used both imported and domestic ST kits, five relied on domestic ST kits only, and one used imported ST kits only, while four centers never set a grey zone for ST throughout the decade. For NAT modalities, eight centers adopted both individual nucleic acid test (ID-NAT) and minipool nucleic acid test (MP-NAT), eight used MP-NAT only and one used ID-NAT only. Seven centers combined transcription mediated amplification (TMA) and polymerase chain reaction (PCR), nine used PCR only and one used TMA only, and fourteen centers ran both imported and domestic NAT systems, two used imported systems only and one used a domestic system only. Over the ten-year period, the mean RR FD across the centers ranged from 2.22 to 12.33 per 10 6 person-years, RR RD from 0.83 to 3.29 per 10 6 person-years and RR TD from 1.59 to 9.29 per 10 6 person-years, with center Z4 consistently showing the lowest values for all three metrics and center U4 recording the highest RR FD and RR TD, while center D2 had the highest RR RD. In 2024 compared with 2015, eleven centers achieved a lower RR FD and ten centers achieved lower RR RD and RR TD. The RR FD and RR TD of centers W2 and U4 displayed pronounced fluctuations and an upward trend in recent years. Conclusions:The 17 provincial blood centers maintain consistent HIV screening strategies, while demonstrating variations in screening algorithm, ST kit manufacturers, NAT modalities, methods, and platform. And the RR FD, RR RD, and RR TD differ across centers. Although most centers show declining trend in RR over the ten-year period, some centers exhibite data fluctuations with a rising trend, suggesting potential for further optimization of HIV screening protocols.

Objective This study reveals the potential mechanism of electroacupuncture in alleviating the inflamma-tory injury of sensory nerves and alleviating the abnormal sensation on the ocular surface in type 2 diabetes rats with dry eye.Methods Healthy male SD rats were induced to establish type 2 diabetes dry eye rat model.Model rats were ran-domly divided into model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group in the 12th week of the experiment.The blank group was established as well,and each group was intervened for 2 weeks.The corneal fluorescein sodium staining(FL),break-up time(BUT),phenol red thread test(PRT),corneal mechanical per-ception threshold(CTT)and blink frequency were measured in each group of rats.After sampling,the expression of neu-ronal injury markers,neuron specific enolase(NSE),neurofilament light chain protein Nf Light(NF-L),phosphorylated microtubule associated protein(Phospho-TAU),as well as inflammatory factors of cysteine protease-1(Caspase-1),inter-leukin-18(IL-18),and interleukin-1β(IL-1β),were observed in the trigeminal ganglion(TG)and spinal cord caudate nu-cleus(SpVc)of each group of rats.Results At the end of the 14th week,compared with the blank group,the FL of the model group,electroacupuncture group,sham-acupuncture group,and fluorometholone group significantly increased,while PRT and BUT significantly decreased.Moreover,the CTT and blink frequency of the model group,sham-acupuncture group,and fluorometholone group significantly decreased(all P＜0.01).Compared with the model group,the electroacu-puncture group and the fluorometholone group showed a decrease in FL,as well as an increase in BUT,PRT,CTT,and blink frequency,with statistically significant differences(all P＜0.05).Compared with the electroacupuncture group,the sham-acupuncture group showed significantly higher FL along with significantly lower values in BUT,PRT,CTT,and blink frequency(all P＜0.05),while the fluorometholone group exhibited decreased FL,PRT,CTT,and blink frequency but in-creased BUT(all P＜0.05).Compared to the sham-acupuncture group,the fluorometholone group exhibited significantly lower FL,but higher BUT and PRT(all P＜0.01).In TG and Sp Vc,compared with the blank group,the expression of NSE,NF-L and Phospho-TAU increased in the model group and the sham-acupuncture group(all P＜0.05);compared with the model group,the expression of NSE,NF-L and Phospho-TAU significantly reduced in the electroacupuncture group(P＜0.01);compared with the electroacupuncture group,the expression of NSE,NF-L and Phospho-TAU increased in the electroacupuncture group and the expression of Phospho-TAU in the sham-acupuncture group also increased(P＜0.05);compared with the sham-acupuncture group,the expression of Phospho-TAU or NF-L significantly reduced in the fluoro-metholone group(all P＜0.05).The Western Blot results indicate that compared with the blank group,the protein expres-sion of Caspase-1,IL-1β,and IL-18 in TG and SpVc of the model group increased;moreover,the expression of Caspase-1 and IL-18 proteins in the sham-acupuncture group also increased(all P＜0.05).Compared with the model group,the ex-pression of Caspase-1,IL-1β,and IL-18 proteins in TG and SpVc reduced in the electroacupuncture group and the fluoro-metholone group,and the expression of IL-1 β proteins in TG and SpVc also reduced in the sham-acupuncture group(all P＜0.05).Compared with the electroacupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc increased in the sham-acupuncture group(all P＜0.05).Compared with the sham-acupuncture group,the expression of Caspase-1,IL-1 β,and IL-18 proteins in TG and SpVc reduced in the fluorometholone group(all P＜0.05).Conclusion Electroacupuncture can effectively alleviate the abnormal ocular surface sensation of type 2 diabetes dry eye rats by alleviating the neuroinflammatory injury in TG and SpVc.

Objective:To analyze the human immunodeficiency virus (HIV) screening status and the resulting residual risk (RR) among blood donors across 17 provincial blood centers in China.Methods:This study used a cross-sectional study. Data on HIV infection markers per 100 000 first-time donors (FD) and repeat donors (RD) from January 2015 to December 2024 were extracted from the National Blood Establishment Performance Comparison Information Management System. Questionnaires were used to collect each center′s HIV screening strategy, algorithm, serological test (ST) kit manufacturers, gray-zone setting for ST, and nucleic acid test (NAT) modality, method, and platform. The incidence-window-period model was used to calculate the residual risk for first-time donors (RR FD), repeat donors (RR RD), and total donors (RR TD) at each center. Horizontal and vertical analysis of RR FD, RR RD, and RR TD across centers and years were performed. Results:All 17 centers applied the same HIV screening strategy which was two rounds of ST followed by one round of NAT. Eight of them operated a single screening algorithm, six employed two algorithms and three used three. Eleven centers used both imported and domestic ST kits, five relied on domestic ST kits only, and one used imported ST kits only, while four centers never set a grey zone for ST throughout the decade. For NAT modalities, eight centers adopted both individual nucleic acid test (ID-NAT) and minipool nucleic acid test (MP-NAT), eight used MP-NAT only and one used ID-NAT only. Seven centers combined transcription mediated amplification (TMA) and polymerase chain reaction (PCR), nine used PCR only and one used TMA only, and fourteen centers ran both imported and domestic NAT systems, two used imported systems only and one used a domestic system only. Over the ten-year period, the mean RR FD across the centers ranged from 2.22 to 12.33 per 10 6 person-years, RR RD from 0.83 to 3.29 per 10 6 person-years and RR TD from 1.59 to 9.29 per 10 6 person-years, with center Z4 consistently showing the lowest values for all three metrics and center U4 recording the highest RR FD and RR TD, while center D2 had the highest RR RD. In 2024 compared with 2015, eleven centers achieved a lower RR FD and ten centers achieved lower RR RD and RR TD. The RR FD and RR TD of centers W2 and U4 displayed pronounced fluctuations and an upward trend in recent years. Conclusions:The 17 provincial blood centers maintain consistent HIV screening strategies, while demonstrating variations in screening algorithm, ST kit manufacturers, NAT modalities, methods, and platform. And the RR FD, RR RD, and RR TD differ across centers. Although most centers show declining trend in RR over the ten-year period, some centers exhibite data fluctuations with a rising trend, suggesting potential for further optimization of HIV screening protocols.

Purpose To analyze and discuss the clinicopathological,molecular pathological characteristics,as well as diagnostic and prognostic features of pleomorphic xanthoastrocytoma(PXA)according to the new WHO classifi-cation.Methods 79 cases of PXA were collected to analyze their pathological and clinical data.Immunohistochemis-try using the EnVision method was employed to detect the expression of CD34,ATRX,Rb,Olig-2,H3K27M,H3K27me3,IDH1 R132H,BRAF VE1 and Ki67.Sanger sequencing was used to detect mutations in H3F3A and IDH1/2.Fluorescence quantitative PCR was used to detect the BRAF V600E mutation and TERT promoter region al-terations.Fluorescence in situ hybridization(FISH)was used to detect CDKN2A and EGFR alterations.The relation-ship between clinical,pathological,molecular genetics data,and prognosis was analyzed.Results The patients'ages ranged from 9 to 69 years,with an average age of 36.4 years.Most tumors were located in the temporal lobe,frontal lobe and parietal lobe.Among the 79 cases,42 were classified as grade 2 PXA and 37 as grade 3 PXA.The tumor cells exhibited pleomorphic changes,with perivascular lymphocytic sheaths and eosinophilic bodies frequently ob-served.Grade 3 PXA exhibited more mitotic figures(average of 11.8/10 HPF),and was usually accompanied by nec-rosis,focal marginal infiltration and microvascular proliferation.Immunohistochemistry and molecular characteristics revealed frequent positivity for CD34,BRAF V600E mutation(68.1％),and CDKN2A homozygous deletion(36.8％)in PXA.Some cases showed TERT gene mutation and absent Rb expression.Univariate survival analysis in-dicated that necrosis,focal marginal infiltration,and CNS WHO grade were related to overall survival,while focal infil-tration and CNS WHO grade were the independent risk factors.Conclusion The prognosis of CNS WHO grade 3 PXA is wrose than that of grade 2 PXA.Accurate diagnosis of PXA requires the combination of the morphological features,immunohistochemical staining,and multiple molecular tests.

Objective To investigate the mechanism by which Erdong Xiaoke Formula(EDXKF)regulates lacrimal gland autophagy in type 2 diabetes-induced dry eye rats through the peroxisome proliferator-activated receptor γ(PPARγ)/mammalian target of rapamycin(mTOR)signal pathway.Methods Healthy male SD rats were fed with high-sugar and high-fat chow and then injected intraperitoneally 30 mg·kg-1 streptozotocin to construct type 2 diabetes-induced dry eye rat models.Healthy male SD rats were selected as a blank group.Type 2 diabetes rats were randomly divided into a model group,a Traditional Chinese Medicine(TCM)group(given 11 g·kg-1 of EDXKF through gavage),an antagonist group(given 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection),and a TCM plus antagonist group(given 11 g·kg-1 EDXKF through gavage and 2 mg·kg-1 PPARγ antagonist through intraperitoneal injection).Fasting blood glucose(FBG),corneal fluorescein(FL)staining,tear film break-up time(BUT)and phenol red cotton thread test(Prtt)were examined before modeling,after modeling,and after intervention.The weight of the lacrimal gland was compared among different groups after sampling.Hematoxylin and eosin(HE)staining was performed to analyze lacrimal gland hismorphol-ogy.The expression of microtubule-associated protein 1 light chain 3(LC3)and Sequestosome-1(p62)in the lacrimal gland was examined by immunofluorescence.Western blot was used to detect the expression of PPARγ,p62,LC3 Ⅱ,LC3 I,mTOR and p-mTOR in the lacrimal gland.Results FBG levels in the antagonist group were significantly increased after intervention,compared with those in the TCM group(P＜0.01).Ocular surface examination showed that compared with the model group,the TCM group had increased BUT and Prtt scores and decreased FL scores,the antagonist group had de-creased BUT and Prtt scores and increased FL scores(all P＜0.05).Compared with the antagonist group,the TCM plus antagonist group showed increased BUT and Prtt scores and decreased FL scores(all P＜0.05).The weight analysis of lac-rimal glands revealed that the lacrimal gland weight increased in the TCM group and decreased in the antagonist group,compared with that in the model group(all P＜0.01).The lacrimal gland weight in the TCM plus antagonist group was higher than that in the antagonist group(P＜0.01).HE staining of the lacrimal gland showed atrophy of glandular lobules,increased fusion and expansion of follicles,and dense distribution of nuclei in model and antagonist groups.These symp-toms were more obvious in the antagonist group.Compared with antagonist and TCM plus antagonist groups,the TCM group showed improved symptoms,with tightly arranged follicles,partial atrophy and fusion,and a small amount of expan-sion.Immunofluorescence staining for detecting the average fluorescence intensity of LC3 and p62 showed that compared with the model group,the TCM group had increased LC3 levels and decreased p62 levels,and the antagonist group had de-creased LC3 levels and increased p62 levels(all P＜0.01).Compared with those in the antagonist group,LC3 levels were increased and p62 levels were decreased in the TCM plus antagonist group(both P＜0.01).Western blot results showed that compared with the model group,the TCM group had increased PPARγ and LC3 levels and decreased p62 and p-mTOR/mTOR levels.PPARγ and LC3 levels were decreased and p62 and p-mTOR/mTOR levels were increased in the antagonist group,compared with those in the model group.The TCM plus antagonist group had higher PPARγ and LC3 levels and low-er p62 and p-mTOR/mTOR levels than the antagonist group(all P＜0.05).Conclusion EDXKF can regulate the PPARy/mTOR signal pathway to promote lacrimal gland autophagy and thus alleviate dry eyes in type 2 diabetes rats.

Objective To observe the effect of electroacupuncture on corneal Toll-like receptor 4(TLR4)-mediated inflammatory signaling pathway in diabetic dry eye rats and to explore the mechanism of electroacupuncture in the treat-ment of diabetic dry eye.Methods A type 2 diabetic rat model was established in 32 healthy male Sprague-Dawley(SD)rats by intraperitoneal injection of streptozotocin(30 mg·kg-1)for 12 weeks after feeding with high-sugar and high-fat di-et for 4 weeks.Twenty-five successfully modeled diabetic dry eye rats were randomly divided into the model group(non-in-tervention),electroacupuncture group(the"Jingming","Cuanzhu","Sizhukong","Taiyang"and"Tongziliao"acupoints were treated with acupuncture,and then"Cuanzhu"and"Tongzilao"acupoints were treated with electroacupuncture,15 min for each time,once a day),sham acupuncture group(blunt-tip needle pricking was performed at the same acupoints as the electroacupuncture group),and fluorometholone group(1 g·L-1fluorometholone eye drops were used in both eyes at 8 o'clock,13 o'clock,and 18 o'clock,1 drop each time),with 6 rats in each group,lasting for 2 weeks.Another 6 healthy male SD rats were selected as a blank group.Random blood glucose,tear film breakup time(BUT),tear secre-tion,corneal fluorescein staining(FL)score,and corneal touch threshold(CTT)of rats in each group were detected be-fore modeling,after modeling,and after the intervention.Hematoxylin and Eosin(HE)staining was performed to observe the corneal morphologic changes in each group.Immunofluorescence histochemical staining was adopted to detect the cor-neal TLR4-positive expression in each group.The TLR4,phosphorylated nuclear factor-kappa P65(P-NF-KB P65),inter-leukin(IL)-1β,and IL-18 expression levels in the cornea were detected by Western blot.Results After modeling,com-pared with the blank group,BUT,tear secretion and CTT decreased and FL increased in all experimental groups,and the differences were statistically significant(all P＜0.01).After the intervention,compared with the model group,FL de-creased,and BUT,tear secretion and CTT increased in the electroacupuncture group,and the differences were statistically significant(all P＜0.05).Corneal HE staining showed that after the intervention,the corneal surface of rats in the model group and sham acupuncture group was not smooth,and the corneal epithelial cells were thickened and disorganized;the corneal surface of rats in the electroacupuncture group and fluorometholone group was smooth,and the corneal epithelial cells were arranged neatly.After the intervention,compared with the blank group,the corneal TLR4 expression of rats in all other groups was elevated,and the differences were statistically significant(all P＜0.05);compared with the model group,the corneal TLR4 expression of rats in the electroacupuncture group and fluorometholone group was reduced(both P＜0.01).Compared with the blank group,corneal TLR4,P-NF-κBP65,IL-1β and IL-18 protein expressions increased in the model group and sham acupuncture group(all P＜0.05);compared with the model group,these expressions decreased in the electroacupuncture group and the fluorometholone group(all P＜0.05).Conclusion Electroacupuncture can im-prove the ocular surface signs and inhibit the expressions of TLR4,P-NF-κB P65,IL-1 β,and IL-18 in the cornea of type 2 diabetic dry eye rats,and its mechanism of action may be related to the regulation of TLR4-mediated inflammatory signaling pathway,which inhibits ocular surface inflammation in diabetic dry eye rats.

Objective To reveal the potential mechanism of electroacupuncture regulating the neuroinflammatory signaling pathway mediated by triggering receptor expressed on myeloid cells 2(TREM2)in trigeminal ganglion(TG)and caudate nucleus of spinal trigeminal nucleus(SpVc)of rats with type 2 diabetes and dry eyes and alleviating ocular surface sensory abnormalities.Methods Healthy male Sprague-Dawley(SD)rats were induced by intraperitoneal injection of 10 g·L-1 streptozotocin to establish a type 2 diabetes dry eye rat model after 4 weeks of high-glucose and high-fat diets.In the 12th week of the experiment,successfully modeled rats were randomly divided into the model group,electroacupunc-ture group,sham-acupuncture group and fluorometholone group,and healthy male SD rats fed with normal diets were se-lected as the blank group.Rats in all groups were intervened for 2 weeks.The corneal fluorescein sodium staining(FL),tear secretion[detected by phenol red thread(PRT)test],tear film break-up time(BUT),and corneal touch threshold(CTT)in each group were measured before,after modeling,and after the intervention.The changes in tissue morphology of TG and SpVc and TREM2 positive expression sites in each group were observed.The messenger ribonucleic acid(mR-NA)expression of TREM2,interleukin-18(IL-18)and interleukin-1β(IL-1β)of TG and Sp Vc in each group were detec-ted.Results After modeling,compared with the blank group,the FL scores of rats significantly increased and PRT,BUT and CTT significantly decreased in other groups(all P＜0.01).After the intervention,compared with the model group,the electroacupuncture group and fluorometholone group showed significant reductions in FL scores and significant increases in PRT,BUT and CTT(all P＜0.01);there was no statistically significant difference in these indicators in the sham-acupuncture group(all P＞0.05).Compared with the electroacupuncture group,the PRT and CTT of rats in the flu-orometholone group and sham-acupuncture group were significantly reduced,the FL score significantly increased and BUT significantly decreased in the sham-acupuncture group(all P＜0.01),and there was no statistically significant difference in FL score and BUT of rats in the fluorometholone group(both P＞0.05).Compared with the sham-acupuncture group,the fluorometholone group showed a decrease in FL score and an increase in PRT,BUT and CTT in rats,with statistically dif-ferent significances(all P＜0.05).The immunofluorescence double-labeling assay showed a positive expression of TREM2 in activated microglia of TG and SpVc in the model group.The reverse transcription-polymerase chain reaction of TG and SpVc showed that compared with the blank group,the TREM2 mRNA expression in the TG and SpVc of the model group,electroacupuncture group,fluorometholone group and sham-acupuncture group decreased,the IL-18 and IL-1β mRNA in TG and SpVc of the model group,IL-18 and IL-1β mRNA in TG and IL-1β mRNA in SpVc of the sham-acupuncture group increased(all P＜0.05);compared with the model group,the TREM2 mRNA expression increased and IL-18 and IL-1β mR-NA expression decreased in the TG and SpVc of the electroacupuncture group and fluorometholone group(all P＜0.05).Compared with the electroacupuncture group,the TREM2 mRNA expression decreased and IL-18,IL-1β mRNA expression increased in TG and SpVc of the sham-acupuncture group(all P＜0.05),and no significant difference was found in TREM2,IL-18 and IL-1β mRNA expression in TG and SpVc of the fluorometholone group(all P＞0.05).Compared with the sham-acupuncture group,the TREM2 mRNA expression in TG and SpVc increased in the fluorometholone group,while the IL-18 mRNA expression in SpVc decreased(all P＜0.05);the IL-1β mRNA in TG and SpVc and the IL-18 mRNA in TG of the fluorometholone group showed no statistically significant difference(all P＞0.05).Conclusion Electroacupunc-ture can effectively alleviate the ocular surface sensory abnormalities of rats with type 2 diabetes and dry eyes by regulating the inflammatory signaling pathway mediated by TREM2 in TG and SpVc.