Objective To establish an efficient and stable one-step real-time quantitative PCR(qRT-PCR)method for detecting mi-croRNA-like small RNAs(milRNAs)encoded by novel Bunyavirus and evaluate its clinical application value.Methods Three kinds of milRNAs encoded by novel Bunyavirus,including sRNA S-1480,sRNA M-692,and sRNA L-4706,were selected based on prelimi-nary screening.The specific qRT-PCR primers and reaction systems for them were designed and optimized.The sensitivity,specificity,and clinical applicability of the method were further evaluated by standard curve drawing,primer concentration/sequence optimization,fluorescent dye optimization,product sequencing verification,and serum specimen detection of 20 patients infected with novel Bunya-virus and 20 healthy controls.Results A one-step qRT-PCR detection system was successfully established.The standard curve con-structed with standard substance showed a good linear relationship in the range of 10 fmol/L to 100 nmol/L(R2>0.98).The detection limits for three kinds of milRNAs were 10 fmol/L.The analysis results of the receiver operating characteristics(ROC)curve showed that sRNA S-1480(AUCROC=0.972 5),sRNA M-692(AUCROC=0.757 5),sRNA L-4706(AUCROC=0.957 5),and their combina-tion(AUCROC=0.995 0)could effectively distinguish the patients infected with novel Bunyavirus from healthy controls.Conclusion The established one-step qRT-PCR detection system exhibits high sensitivity and provides an efficient and reliable alternative for the clinical diagnosis of novel Bunyavirus infection.

Objective To investigate preventive effect of Huanshao capsule on postoperative re-currence of moderate to severe intrauterine adhesion and its impact on postoperative pregnancy rates.Methods A total of 163 patients with moderate to severe intrauterine adhesion after surgery were se-lected as study subjects,and randomly divided into observation group(n=82)and control group(n=81).The control group received conventional treatment,while the observation group additionally took Huanshao capsule postoperatively.Serum fibronectin(FN),laminin(LN),vascular endothelial growth factor(VEGF)and transforming growth factor-β1(TGF-β1)levels were compared between two groups at 1 month,3 and 6 months postoperatively.Endometrial thickness,uterine artery blood flow resistance index(RI),postoperative pregnancy rates and quality of life scores were also com-pared.Results At 3 and 6 months postoperatively,serum FN,LN and VEGF levels in the observa-tion group were lower than those in the control group(P＜0.05).At 6 months postoperatively,serum TGF-β1 in the observation group was lower than that in the control group,with statistically significant difference(P＜0.05).At 3 and 6 months after surgery,endometrial thickness in the observation group was significantly higher,uterine artery flow RI was significantly lower than,and scores of various quality of life(physiological function,physical pain,social function and mental health)were signifi-cantly higher than those in the control group(P＜0.05).At 6 and 9 months postoperatively,the pregnancy rate in the observation group was higher than that in the control group,with statistically sig-nificant differences(P＜0.05).Conclusion Huanshao capsule can significantly reduce serum FN,LN,VEGF and TGF-β1 levels in patients with moderate to severe intrauterine adhesion after surgery,improve the postoperative endometrial environment,and increase postoperative pregnancy rates.

Objective:To compare the efficacy of pars plana vitrectomy (PPV) combined with subretinal or intravitreal injection of Conbercept for the treatment of refractory diabetic macular edema (DME).Methods:A retrospective case control study. From June 2022 to March 2024, 32 eyes of 32 patients with refractory DME diagnosed at The Affiliated Eye Hospital of Nanchang University were included in the study. There were 17 male cases with 17 eyes and 15 female cases with 15 eyes. Age was (57.44±8.99) years old; The duration of diabetes was (12.72±6.11) years. All patients had received regular treatment with anti-vascular endothelial growth factor (VEGF) drugs or corticosteroid drugs for at least 5 times, and had undergone focal retinal laser photocoagulation or panretinal laser photocoagulation, the central macular thickness (CMT) persisted or decreased by less than 50 μm. All affected eyes underwent best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), microperimetry, and laboratory glycated hemoglobin (HbA1c) testing. BCVA was measured using a standard logarithmic visual acuity chart, and converted to the logarithm of the minimum angle of resolution (logMAR) for statistical analysis. CMT was measured using an OCT device. Microperimetry was performed using an MP-3 microperimeter, recording the mean sensitivity (MS) of the retina within a 12° range of the fovea. The affected eyes were treated with 23G PPV combined with internal limiting membrane peeling and either macular subretinal or intravitreal injection of Conbercept, and were divided into subretinal injection group and the intravitreal injection group, each consisting of 16 cases and 16 eyes. The same equipment and methods as before surgery were used for related examinations at 1, 3, and 6 months post-surgery. Changes in BCVA, CMT, and MS were observed and compared, as well as the number of additional anti-VEGF treatments required within 6 months after surgery. Intergroup comparisons were made using independent samples t tests, and repeated measures data were analyzed using repeated measures analysis of variance. Results:The age ( t=-0.271), gender composition ( χ2=0.001), duration of diabetes ( Z=-0.868), HbA1c ( t=-0.789), intraocular pressure ( t=1.689), logMAR BCVA ( t=1.393), CMT ( t=-0.613), MS ( Z=-0.132), and the number of anti-VEGF injections ( t=-0.752) between the subretinal injection group and the intravitreal injection group showed no statistically significant differences ( P>0.05). The within-subject effects comparison of BCVA, CMT, and MS at 1, 3, and 6 months post-surgery compared to pre-surgery for all affected eyes showed statistically significant differences ( F=8.060, 125.722, 39.054; P<0.05). The overall comparison of logMAR BCVA between the subretinal and intravitreal injection groups post-surgery showed no statistically significant difference ( F=0.662, P=0.422), however, comparisons of CMT ( F=4.540) and MS ( F=6.066) showed statistically significant differences ( P<0.05). At 1, 3, and 6 months post-surgery, comparisons of logMAR BCVA between the two groups showed no statistically significant differences ( t=-0.123, 0.239, 1.087; P>0.05), comparisons of CMT showed statistically significant differences ( t=-3.474, -4.832, -2.482; P<0.05), comparisons of MS showed statistically significant differences at 1 and 3 months ( t=-2.940, -2.545; P<0.05), but not at 6 months ( t=-1.527, P>0.05). At 6 months post-surgery, the number of additional intravitreal anti-VEGF injections required in the subretinal and intravitreal injection groups showed a statistically significant difference ( Z=-2.033, P=0.042). During the follow-up period and at the final follow-up, no complications such as injection site bleeding, retinal detachment, vitreous hemorrhage, macular hole, or retinal pigment epithelial tear or atrophy occurred in all affected eyes. Conclusion:Compared with intravitreal injection, subretinal injection of Conbercept for the treatment of refractory DME has more advantages in reducing macular edema and improving visual function in the macular area, and also reduces the number of postoperative anti-VEGF drug treatments.

Objective:To compare and observe the efficacy and safety of pars plana vitrectomy (PPV) combined with 41G ultrafine needle injection of balanced salt solution (BSS) and internal limiting membrane inversion and coverage in the treatment of large-diameter macular hole (MH).Methods:A prospective study. From April 2023 to April 2024, 42 patients (42 eyes) diagnosed with large-diameter MH at The Affiliated Eye Hospital of Nanchang University were included in the study. The substrate diameters (BD) of MH are all greater than 1 000 μm. All affected eyes underwent best corrected visual acuity (BCVA), microvisual field, optical coherence tomography (OCT), and OCT angiography (OCTA) examinations before surgery. BCVA examination was conducted using the international standard logarithmic visual acuity chart, and the statistics were converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity. Microfield-of-view examination records the mean retinal sensitivity (MS) within a 12° range of the fovea. The minimum diameter (MD) and BD of the hole were measured by OCT and the MH index (MHI) was calculated. OCTA measures the area and perimeter (PERIM) of the foveal avascular zone (FAZ) in the fovea centralis, as well as the retinal vessel length density (VLD) and vessel perfusion density (VPD) in the central area. The affected eyes were divided into the observation group (22 eyes, treated with PPV combined with 41G ultra-micro needle subretinal injection of BSS and the control group (20 eyes, treated with PPV combined with internal limiting membrane inversion and coverage) according to the random number table method. The patients were followed up for 6 months after the operation, and the above indicators were reexamined at 1, 3 and 6 months. The changes of BCVA, MS, FAZ parameters, VLD, VPD before and after the operation, as well as the closure of the hole and the occurrence of complications were compared between the two groups. The independent sample t test was used for comparison between groups. The correlation between postoperative indicators and preoperative structural parameters was analyzed using Pearson analysis. Results:In the 22 eyes of the observation group, the hole closure rate was 100.0% (22/22) 6 months after the operation, and the complete closure rate was 90.9% (20/22). In the control group of 20 eyes, the closure rate was 95.0% (19/20), and the complete closure rate was 90.0% (18/20). The hole closure rate of the affected eyes in the observation group was better than that in the control group, but the difference was not statistically significant ( χ2=1.736, P=0.420). The logMAR BCVA, MS, FAZ area, PERIM, VLD and VPD at each time point after the operation in both groups were significantly improved compared with those before the operation, and the differences were statistically significant ( P<0.05). The results of Pearson correlation analysis showed that in the observation group, logMAR BCVA at 6 months after surgery was positively correlated with BD before surgery ( r=0.340, P=0.029), and negatively correlated with MHI before surgery ( r=?0.350, P=0.023). Six months after the operation, MS was positively correlated with the preoperative hole height and MHI ( r=0.330, P=0.034). In the control group, 6 months after the operation, MS was negatively correlated with BD before the operation ( r=?0.480, P=0.032), and positively correlated with MHI before the operation ( r=0.510, P=0.027). Six months after the operation, the FAZ area of the affected eyes in the observation group [(0.17±0.09) mm 2] was larger than that in the control group [(0.12±0.07) mm 2], and the difference was statistically significant ( t=?0.340, P=0.025). No complications such as abnormal intraocular pressure, retinal pigment epithelium injury or retinal detachment occurred in all the affected eyes after the surgery. Conclusions:PPV combined with 41G ultra-micro needle subretinal injection of BSS for the treatment of large-diameter MH has a high rate of hole closure and good safety. Larger BD and MD, lower MHI and hole height before surgery may affect the recovery of visual function after surgery.

Objective:To analyze the clinical characteristics and identify predictive factors for spontaneous passage of common bile duct (CBD) stones.Methods:A retrospective analysis was conducted on patients diagnosed as having choledocholithiasis via abdominal imaging at outpatient and emergency departments of Jiangsu Province Hospital of Chinese Medicine and other medical institutions from January 2021 to November 2023. Participants were stratified into spontaneous passage versus non-passage groups. Multivariate logistic regression analysis was used to identify predictors for the spontaneous passage of common bile duct stones.Results:Spontaneous stone passage were confirmed in 70 cases (15.5%). Multivariate logistic regression analysis identified that an admission diagnosis of acute biliary pancreatitis ( OR=3.317, 95% CI: 1.427-7.713, P=0.005), larger common bile duct diameter ( OR=1.117, 95% CI: 1.000-1.248, P=0.049), and solitary stones ( OR=11.135, 95% CI: 3.602-34.418, P<0.001) significantly increased the probability of spontaneous stones. In contrast, larger stone long diameter ( OR=0.539, 95% CI: 0.441-0.659, P<0.001) markedly decreased passage likelihood. Receiver operator characteristic (ROC) curve analysis demonstrated that the common bile duct diameter predicted spontaneous stone passage with an area under the curve (AUC) of 0.662, yielding sensitivity of 52.9% (37/70) and specificity of 73.6% (51/70) at a cutoff value of 9.5 mm. The common bile duct stone diameter achieved an AUC of 0.852 for predicting spontaneous stone passage, with sensitivity of 75.7% (53/70) and specificity of 89.0% (62/70) at a cutoff value of 4.5 mm. Conclusion:Solitary small stones, ductal dilation, and an admission diagnosis of acute biliary pancreatitis are key predictive factors for spontaneous common bile duct stone passage. A common bile duct diameter ≥9.5 mm and stone long diameter ≤4.5 mm are more likely to result in spontaneous stone passage.

Objective To establish an efficient and stable one-step real-time quantitative PCR(qRT-PCR)method for detecting mi-croRNA-like small RNAs(milRNAs)encoded by novel Bunyavirus and evaluate its clinical application value.Methods Three kinds of milRNAs encoded by novel Bunyavirus,including sRNA S-1480,sRNA M-692,and sRNA L-4706,were selected based on prelimi-nary screening.The specific qRT-PCR primers and reaction systems for them were designed and optimized.The sensitivity,specificity,and clinical applicability of the method were further evaluated by standard curve drawing,primer concentration/sequence optimization,fluorescent dye optimization,product sequencing verification,and serum specimen detection of 20 patients infected with novel Bunya-virus and 20 healthy controls.Results A one-step qRT-PCR detection system was successfully established.The standard curve con-structed with standard substance showed a good linear relationship in the range of 10 fmol/L to 100 nmol/L(R2>0.98).The detection limits for three kinds of milRNAs were 10 fmol/L.The analysis results of the receiver operating characteristics(ROC)curve showed that sRNA S-1480(AUCROC=0.972 5),sRNA M-692(AUCROC=0.757 5),sRNA L-4706(AUCROC=0.957 5),and their combina-tion(AUCROC=0.995 0)could effectively distinguish the patients infected with novel Bunyavirus from healthy controls.Conclusion The established one-step qRT-PCR detection system exhibits high sensitivity and provides an efficient and reliable alternative for the clinical diagnosis of novel Bunyavirus infection.

Objective:To compare the efficacy of pars plana vitrectomy (PPV) combined with subretinal or intravitreal injection of Conbercept for the treatment of refractory diabetic macular edema (DME).Methods:A retrospective case control study. From June 2022 to March 2024, 32 eyes of 32 patients with refractory DME diagnosed at The Affiliated Eye Hospital of Nanchang University were included in the study. There were 17 male cases with 17 eyes and 15 female cases with 15 eyes. Age was (57.44±8.99) years old; The duration of diabetes was (12.72±6.11) years. All patients had received regular treatment with anti-vascular endothelial growth factor (VEGF) drugs or corticosteroid drugs for at least 5 times, and had undergone focal retinal laser photocoagulation or panretinal laser photocoagulation, the central macular thickness (CMT) persisted or decreased by less than 50 μm. All affected eyes underwent best-corrected visual acuity (BCVA), intraocular pressure, optical coherence tomography (OCT), microperimetry, and laboratory glycated hemoglobin (HbA1c) testing. BCVA was measured using a standard logarithmic visual acuity chart, and converted to the logarithm of the minimum angle of resolution (logMAR) for statistical analysis. CMT was measured using an OCT device. Microperimetry was performed using an MP-3 microperimeter, recording the mean sensitivity (MS) of the retina within a 12° range of the fovea. The affected eyes were treated with 23G PPV combined with internal limiting membrane peeling and either macular subretinal or intravitreal injection of Conbercept, and were divided into subretinal injection group and the intravitreal injection group, each consisting of 16 cases and 16 eyes. The same equipment and methods as before surgery were used for related examinations at 1, 3, and 6 months post-surgery. Changes in BCVA, CMT, and MS were observed and compared, as well as the number of additional anti-VEGF treatments required within 6 months after surgery. Intergroup comparisons were made using independent samples t tests, and repeated measures data were analyzed using repeated measures analysis of variance. Results:The age ( t=-0.271), gender composition ( χ2=0.001), duration of diabetes ( Z=-0.868), HbA1c ( t=-0.789), intraocular pressure ( t=1.689), logMAR BCVA ( t=1.393), CMT ( t=-0.613), MS ( Z=-0.132), and the number of anti-VEGF injections ( t=-0.752) between the subretinal injection group and the intravitreal injection group showed no statistically significant differences ( P>0.05). The within-subject effects comparison of BCVA, CMT, and MS at 1, 3, and 6 months post-surgery compared to pre-surgery for all affected eyes showed statistically significant differences ( F=8.060, 125.722, 39.054; P<0.05). The overall comparison of logMAR BCVA between the subretinal and intravitreal injection groups post-surgery showed no statistically significant difference ( F=0.662, P=0.422), however, comparisons of CMT ( F=4.540) and MS ( F=6.066) showed statistically significant differences ( P<0.05). At 1, 3, and 6 months post-surgery, comparisons of logMAR BCVA between the two groups showed no statistically significant differences ( t=-0.123, 0.239, 1.087; P>0.05), comparisons of CMT showed statistically significant differences ( t=-3.474, -4.832, -2.482; P<0.05), comparisons of MS showed statistically significant differences at 1 and 3 months ( t=-2.940, -2.545; P<0.05), but not at 6 months ( t=-1.527, P>0.05). At 6 months post-surgery, the number of additional intravitreal anti-VEGF injections required in the subretinal and intravitreal injection groups showed a statistically significant difference ( Z=-2.033, P=0.042). During the follow-up period and at the final follow-up, no complications such as injection site bleeding, retinal detachment, vitreous hemorrhage, macular hole, or retinal pigment epithelial tear or atrophy occurred in all affected eyes. Conclusion:Compared with intravitreal injection, subretinal injection of Conbercept for the treatment of refractory DME has more advantages in reducing macular edema and improving visual function in the macular area, and also reduces the number of postoperative anti-VEGF drug treatments.

Objective:To compare and observe the efficacy and safety of pars plana vitrectomy (PPV) combined with 41G ultrafine needle injection of balanced salt solution (BSS) and internal limiting membrane inversion and coverage in the treatment of large-diameter macular hole (MH).Methods:A prospective study. From April 2023 to April 2024, 42 patients (42 eyes) diagnosed with large-diameter MH at The Affiliated Eye Hospital of Nanchang University were included in the study. The substrate diameters (BD) of MH are all greater than 1 000 μm. All affected eyes underwent best corrected visual acuity (BCVA), microvisual field, optical coherence tomography (OCT), and OCT angiography (OCTA) examinations before surgery. BCVA examination was conducted using the international standard logarithmic visual acuity chart, and the statistics were converted to the logarithm of the minimum angle of resolution (logMAR) visual acuity. Microfield-of-view examination records the mean retinal sensitivity (MS) within a 12° range of the fovea. The minimum diameter (MD) and BD of the hole were measured by OCT and the MH index (MHI) was calculated. OCTA measures the area and perimeter (PERIM) of the foveal avascular zone (FAZ) in the fovea centralis, as well as the retinal vessel length density (VLD) and vessel perfusion density (VPD) in the central area. The affected eyes were divided into the observation group (22 eyes, treated with PPV combined with 41G ultra-micro needle subretinal injection of BSS and the control group (20 eyes, treated with PPV combined with internal limiting membrane inversion and coverage) according to the random number table method. The patients were followed up for 6 months after the operation, and the above indicators were reexamined at 1, 3 and 6 months. The changes of BCVA, MS, FAZ parameters, VLD, VPD before and after the operation, as well as the closure of the hole and the occurrence of complications were compared between the two groups. The independent sample t test was used for comparison between groups. The correlation between postoperative indicators and preoperative structural parameters was analyzed using Pearson analysis. Results:In the 22 eyes of the observation group, the hole closure rate was 100.0% (22/22) 6 months after the operation, and the complete closure rate was 90.9% (20/22). In the control group of 20 eyes, the closure rate was 95.0% (19/20), and the complete closure rate was 90.0% (18/20). The hole closure rate of the affected eyes in the observation group was better than that in the control group, but the difference was not statistically significant ( χ2=1.736, P=0.420). The logMAR BCVA, MS, FAZ area, PERIM, VLD and VPD at each time point after the operation in both groups were significantly improved compared with those before the operation, and the differences were statistically significant ( P<0.05). The results of Pearson correlation analysis showed that in the observation group, logMAR BCVA at 6 months after surgery was positively correlated with BD before surgery ( r=0.340, P=0.029), and negatively correlated with MHI before surgery ( r=?0.350, P=0.023). Six months after the operation, MS was positively correlated with the preoperative hole height and MHI ( r=0.330, P=0.034). In the control group, 6 months after the operation, MS was negatively correlated with BD before the operation ( r=?0.480, P=0.032), and positively correlated with MHI before the operation ( r=0.510, P=0.027). Six months after the operation, the FAZ area of the affected eyes in the observation group [(0.17±0.09) mm 2] was larger than that in the control group [(0.12±0.07) mm 2], and the difference was statistically significant ( t=?0.340, P=0.025). No complications such as abnormal intraocular pressure, retinal pigment epithelium injury or retinal detachment occurred in all the affected eyes after the surgery. Conclusions:PPV combined with 41G ultra-micro needle subretinal injection of BSS for the treatment of large-diameter MH has a high rate of hole closure and good safety. Larger BD and MD, lower MHI and hole height before surgery may affect the recovery of visual function after surgery.

Objective:To analyze the clinical characteristics and identify predictive factors for spontaneous passage of common bile duct (CBD) stones.Methods:A retrospective analysis was conducted on patients diagnosed as having choledocholithiasis via abdominal imaging at outpatient and emergency departments of Jiangsu Province Hospital of Chinese Medicine and other medical institutions from January 2021 to November 2023. Participants were stratified into spontaneous passage versus non-passage groups. Multivariate logistic regression analysis was used to identify predictors for the spontaneous passage of common bile duct stones.Results:Spontaneous stone passage were confirmed in 70 cases (15.5%). Multivariate logistic regression analysis identified that an admission diagnosis of acute biliary pancreatitis ( OR=3.317, 95% CI: 1.427-7.713, P=0.005), larger common bile duct diameter ( OR=1.117, 95% CI: 1.000-1.248, P=0.049), and solitary stones ( OR=11.135, 95% CI: 3.602-34.418, P<0.001) significantly increased the probability of spontaneous stones. In contrast, larger stone long diameter ( OR=0.539, 95% CI: 0.441-0.659, P<0.001) markedly decreased passage likelihood. Receiver operator characteristic (ROC) curve analysis demonstrated that the common bile duct diameter predicted spontaneous stone passage with an area under the curve (AUC) of 0.662, yielding sensitivity of 52.9% (37/70) and specificity of 73.6% (51/70) at a cutoff value of 9.5 mm. The common bile duct stone diameter achieved an AUC of 0.852 for predicting spontaneous stone passage, with sensitivity of 75.7% (53/70) and specificity of 89.0% (62/70) at a cutoff value of 4.5 mm. Conclusion:Solitary small stones, ductal dilation, and an admission diagnosis of acute biliary pancreatitis are key predictive factors for spontaneous common bile duct stone passage. A common bile duct diameter ≥9.5 mm and stone long diameter ≤4.5 mm are more likely to result in spontaneous stone passage.

Objective:To analyze the clinical phenotype and genetic characteristics of a patient with Isidor-Toutain spinal epiphyseal dysplasia (SEMD) due to variant of RPL13 gene. Methods:A pregnant woman at 18 weeks of gestation who had presented at Quzhou Maternal and Child Health Care Hospital on January 14, 2023 was selected as the study subject. Whole exome sequencing (WES) was carried out for the patient, and candidate variant was validated by Sanger sequencing and bioinformatic analysis.Results:The woman was 37 years old with extremely short stature (135 cm) and "O" shaped legs. WES revealed that she has harbored a c. 548G>C (p.Arg183Pro) missense variant of the RPL13 gene (NM_000977.4). The same variant was not found in her fetus. Based on the guidelines from the American College of Medical Genetics and Genomics (ACMG), the variant was predicted to be likely pathogenic (PS4+ PM2_Supporting+ PP3+ PP4). Conclusion:Isidor-Toutain type SEMD due to variants of the RPL13 gene may have variable expressivity and diverse clinical phenotypes. Above finding has facilitated the differential diagnosis and genetic counseling for this family.