Objective To investigate the predictive value of serum lactate dehydrogenase(LDH)level com-bined with high-throughput sequencing in children with severe Mycoplasma pneumoniae pneumonia(SMPP).Methods The clinical data of 99 children with Mycoplasma pneumoniae pneumonia(MPP)admitted to Hua-zhong University of Science and Technology Union Hospital from October 2023 to March 2024 were retro-spectively analyzed.According to the severity of the disease,the children were divided into mild group(33 ca-ses)and severe group(66 cases).Spearman correlation analysis was used for correlation analysis,Logistic re-gression was used to analyze the influencing factors of SMPP,and the receiver operating characteristic(ROC)curve was used to evaluate the value of LDH in predicting SMPP.Results The level of LDH in the severe group was higher than that in the mild group(P<0.05).The results of high-throughput sequencing showed that the proportions of virulence positive and drug resistance gene A2063G mutation in the severe group were higher than those in the mild group(P<0.05).Multivariate Logistic regression analysis showed that LDH level and drug resistance gene A2063G mutation were related to the SMPP(P<0.05).ROC curve analysis showed that LDH combined with high-throughput sequencing detection of drug resistance gene A2063G muta-tion had a high value in predicting SMPP,and the area under the curve was 0.724.Conclusion Serum LDH combined with high-throughput sequencing detection of drug resistance gene A2063G mutation can be used as an effective indicator to predict SMPP.

Medical ultrasound technology is widely used for diagnosis and therapy in clinical practice.Ultrasound probes,which are directly contact with patients,pose a potential risk of pathogen transmission.This expert consen-sus was developed by a multidisciplinary team based on international guidelines,standards in China,and the results of a national survey,aiming to reduce the risk of healthcare-associated infection through standardizing reprocessing of medical ultrasound probes,and formulating consensus recommendations with the Delphi method.The consensus clarifies the reprocessing principles for three types of ultrasound probes of different infection risks:external-use ul-trasound probes,interventional percutaneous ultrasound probes,and internal-use ultrasound probes,puts forward systematic suggestions on the reprocessing standards and disinfection levels of ultrasound probe isolation covers and coupling agents,the reprocessing procedures and methods of ultrasound probes,as well as architectural layout and management of reprocessing,so as to provide a scientific prevention and control framework for ensuring ultrasound diagnosis and therapy safety.

Forty adult male C57BL/6 mice were randomly divided into four groups:the control group,Salmonella typhimurium(S.typhimurium)(ST)group,phlorizin(PHZ)+S.typhimuri-um(ST)group,and PHZ(80 mg/kg)group,with 10 mice in each group.Morphological observa-tion,HE staining,ELISA,immunofluorescence and Western blot were performed,the results showed that PHZ significantly increased the cecal index,decreased the spleen index of S.typhi-murium-induced mice(P＜0.05),and reduce the pathological damage of cecum in mice.Mean-while,PHZ treatment also significantly reduced colonization of S.typhimurium in the cecum,spleen,mesenteric lymph nodes and liver(P＜0.05).The results of ELISA showed that PHZ treatment also significantly inhibited the S.ty phimurium-induced increase in the expression of IL-1β,INF-γ,TNF-a and IL-6 in the cecum of mice(P＜0.05).Immunofluorescence and Western blot results showed that PHZ significantly increased the protein expression levels of Occludin,Claudin-3,and ZO-1 in the cecal barrier of mice induced by S.typhimurium(P＜0.05).These results con-firmed that phlorizin could improve cecal inflammation and intestinal barrier damage induced by S.typhimurium in mice.

Medical ultrasound technology is widely used for diagnosis and therapy in clinical practice.Ultrasound probes,which are directly contact with patients,pose a potential risk of pathogen transmission.This expert consen-sus was developed by a multidisciplinary team based on international guidelines,standards in China,and the results of a national survey,aiming to reduce the risk of healthcare-associated infection through standardizing reprocessing of medical ultrasound probes,and formulating consensus recommendations with the Delphi method.The consensus clarifies the reprocessing principles for three types of ultrasound probes of different infection risks:external-use ul-trasound probes,interventional percutaneous ultrasound probes,and internal-use ultrasound probes,puts forward systematic suggestions on the reprocessing standards and disinfection levels of ultrasound probe isolation covers and coupling agents,the reprocessing procedures and methods of ultrasound probes,as well as architectural layout and management of reprocessing,so as to provide a scientific prevention and control framework for ensuring ultrasound diagnosis and therapy safety.

Forty adult male C57BL/6 mice were randomly divided into four groups:the control group,Salmonella typhimurium(S.typhimurium)(ST)group,phlorizin(PHZ)+S.typhimuri-um(ST)group,and PHZ(80 mg/kg)group,with 10 mice in each group.Morphological observa-tion,HE staining,ELISA,immunofluorescence and Western blot were performed,the results showed that PHZ significantly increased the cecal index,decreased the spleen index of S.typhi-murium-induced mice(P＜0.05),and reduce the pathological damage of cecum in mice.Mean-while,PHZ treatment also significantly reduced colonization of S.typhimurium in the cecum,spleen,mesenteric lymph nodes and liver(P＜0.05).The results of ELISA showed that PHZ treatment also significantly inhibited the S.ty phimurium-induced increase in the expression of IL-1β,INF-γ,TNF-a and IL-6 in the cecum of mice(P＜0.05).Immunofluorescence and Western blot results showed that PHZ significantly increased the protein expression levels of Occludin,Claudin-3,and ZO-1 in the cecal barrier of mice induced by S.typhimurium(P＜0.05).These results con-firmed that phlorizin could improve cecal inflammation and intestinal barrier damage induced by S.typhimurium in mice.

【Objective】 To explore the model of treatment of refractory wounds by apheresis platelet-rich plasma (PRP), and evaluate its efficacy and safety. 【Methods】 PRP treatment was carried out for 34 patients with refractory wounds who were hospitalized in our hospital from June 2020 to December 2023. The patient′s autologous PRP was collected by the blood composition separator. PRP point-like injection and/or direct application along the edge of the wound, or platelet gel (PG) to cover the wound and fill the sinus were used to treat different types of wounds. The frequency of treatment was once to twice a week, with each course of treatment consisting of 4 to 5 sessions. The efficacy was observed and recorded, and the safety was evaluated. 【Results】 Before PRP collection, the basal platelet, white blood cell and red blood cell counts were (321.85±114.64) ×10⁹/L, (9.52±3.21) ×10⁹/L and (4.34±0.62) ×10¹²/L, respectively. The counts of platelets, white blood cells and red blood cells in PRP products were (1 438.53±376.89)×10⁹/L, (1.38±1.03)×10⁹/L and (0.03±0.01)×10¹²/L, respectively. The platelet concentration of PRP products increased by 4.47 times on average, and the sampled bacterial culture was negative. Out of 34 patients treated with PRP, 33 showed improvement in symptoms, while 1 did not respond well due to long-term bedridden and poor compliance. Four patients had mild adverse reactions during PRP collection or treatment, but all were able to complete PRP collection or treatment after standardized treatment. 【Conclusion】 Apheresis PRP products have the advantages of stable quality, safety, reliability and traceability, and can effectively promote the healing of a variety of refractory wounds. Its treatment process is safe and effective, and is worthy of popularization and application.

【Objective】 To explore the effect of autologous platelet-rich plasma(PRP) in the treatment of dry eye syndrome. 【Methods】 A total of 64 patients with dry eye treated in the Ophthalmology Department of Jingdong Sino-American Hospital of Hebei Province from March to November 2023 were randomly selected for the study. The patients were divided into observation group and control group according to the random number table method, with 32 cases and 64 eyes in each group. The control group was treated with artificial tears of polyvinyl alcohol without preservatives, and the observation group was treated with PRP eye drops. The clinical efficacy and adverse reactions of the two groups were compared. Overall efficacy, duration of symptom resolution, ocular surface disease index (OSDI), tear film break up time (TBUT), schirmer's test(St) and corneal fluorescein staining (CFS) results of the two groups were observed and compared in 4 weeks. 【Results】 After 4 weeks of treatment, the total effective rate of observation group was 96.88%(31/32), which was significantly higher than that of control group 84.38%(27/32), with statistical significance (P<0.05). After treatment, the St, TBUT, CFS, OSDI and lacrimal river height in observation group were significantly superior to control group, with statistical significance (P<0.05). The disappearance time of dryness, visual fatigue and foreign body sensation in the observation group was shorter than that in the control group, and the difference was statistically significant (P<0.05). The incidence of adverse reactions in the observation group was 46.875%(15/32), and there was no significant difference between the observation group and the control group. 【Conclusion】 Autologous PRP is an effective and safe choice for the treatment of dry eye syndrome, which is worthy of further study and confirmation.

【Objective】 To investigate the effects of different anticoagulants on platelet-rich plasma(PRP) release content of growth factor and injection pain. 【Methods】 A total of 15 voluntary blood donors were selected, with each blood donor using four kinds of anticoagulant tubes with EDTA-K2 anticoagulation, EDTA-NA2 anticoagulation, citrate anticoagulation, ACD-A anticoagulation respectively as group A, B, C and D. PRP was isolated and prepared by the rich plasma method, and the contents of PDGF-AA, TGF-β, IGF-1, VEGF, and PF-4 were detected by enzyme-linked immunosorbent assay. Meanwhile, SD rats (20, 4 / group) were injected subcutaneously or intradermally with the supernatant of PRP and PG gel prepared in the 4 groups and normal saline in the control group. The pain status of SD rats during the injection was observed and recorded. The pain status of the 5 groups of experimental animals was evaluated according to the American Laboratory Animal Pain Guide. 【Results】 The platelet counts in PRP in group D was the highest [(1 294.53±277.37) × 10⁹/L], which was significantly higher than that in group A [ (789.13±377.13) ×10⁹/L] and group C [ (990.94±493.12) ×10⁹/L] (P<0.05). The OD value of PDGF-AA in group A, B, C, and D were 1.51± 0.18, 1.69±0.21, 0.66±0.19and 1.72±0.13, respectively, with statistically significant difference between groups (P<0.05 ) and group D better than the other three groups. The OD value of PF-4 was 1.18±0.24, 1.61±0.14, 0.65±0.26 and 1.72±0.10 respectively, with statistically significant difference between groups (P<0.05) and group D better than other three groups. The OD value of IGF-1 was 1.02±0.08, 0.98±0.11, 1.06±0.11 and 1.32±0.65 respectively, with no significant difference between groups (P>0.05). The OD value of VEGF was 0.13±0.04, 0.21±0.14, 0.08±0.02 and 0.13±0.04 respectively, with statistically significant difference between group B and C (P<0.05). The OD value of TGF-β was 0.14±0.01, 0.15±0.01, 0.28±0.17 and 1.10±0.37 respectively, with statistically significant difference between groups (P<0.05) and group D better than other three groups. Comparison of injection pain: when the supernatant of PRP and PG gel was injected, there were significant differences between group A, B, C and D, and the control group (P<0.05) . The median pain scores of PRP injection of group A, B, C, and D were 6 (1.5), 5 (0.75), 4.5 (2.5), and 3(3) respectively, with group D lower than other three groups, and no statistically significant difference was noticed (P>0.05) . The median pain scores of the PG supernatant injection of group A, B, C, and D were 4 (2.25), 3 (2.75), 4 (3), 1 (1.5), and the difference was not statistically significant (P>0.05). There was no significant difference between the PRP injection group and the PG supernatant group (P> 0.05). 【Conclusion】 PRP prepared by two-step centrifugation with ACD-A anticoagulant can obtain the higher platelet counts and the maximum release of PDGF-AA, PF-4, IGF-1, and TGF-β. In terms of pain, ACD-A anticoagulant injection has the lowest pain with the animals.

【Objective】 To develop a spray-on membrane dressing for wound repair containing platelet rich plasma (PRP) sodium alginate (SA)/agarose(AG)/carboxymethyl chitosan (CMCS). 【Methods】 SA/AG/ CMCS were mixed in different proportions to prepare biodegradable quick setting spray (BQSS) by blending film method, and the film-forming time, moisture retention and compression resistance of the prepared BQSS were tested. Then PRP and BQSS were mixed in the proportion of 3∶7, 4∶6, 5∶5, 6∶4 and 7∶3 to prepare PRP-BQSS spray film dressings. The film-forming time, moisture retention, compressive strength, porosity and slow-release effect of growth factors of PRP-BQSS spray film dressings were studied. 【Results】 In the preparation of BQSS compound spray film solution, when SA, AG, CMCS and sterile distilled water were 0.6∶0.6∶0.6∶98.2g, the film-forming time (7.73±0.31) s, moisture retention (75. 54±3.03) % and compression resistance (791.00±68.02) g of the spray-film dressing were the best. The basic properties of PRP-BQSS spray-on film dressings and the release of growth factors show that PRP-BQSS spray-on film dressings can exist in different forms, and with the decrease of PRP concentration percentage, its film-forming time, moisturizing performance and compressive strength showed an upward trend. When the PRP content is 30%, the porosity of the dressing is the highest, about(84.34±0.90)%. The release of platelet-derived growth factor-AA(PDGF-AA), platelet factor-4(PF-4) and transforming growth factor beta (TGF-β) was in a slow upward trend, and the release of the three growth factors was higher than that of PRP group in 48 hours. 【Conclusion】 The preparation method of PRP-BQSS spray film dressing designed in this study is simple and mild, and can form a film quickly, with good biological properties and better growth factor inhibition and sustained-release effect.

Objective:To prepare specific anti- Fusarium yolk immunoglobulin (IgY) and investigate its tolerance to temperature and pH and verify its antifungal effect against Fusarium. Methods:Eighteen 22-week-old Leghorn laying hens were selected and randomized into negative control group and experimental group according to the random number table method, with 9 hens in each group.The 2×10 7 colony forming units (CFU)/ml suspension of inactivated hyphae of Fusarium and Freund complete adjuvant was mixed in a 1∶1 ratio and emulsified.The hens in the experimental group were injected with 1 ml of the mixture to immunize and received 1 ml of Freund incomplete adjuvant as booster injection at two weeks after the initial injection.The egg yolk was collected from the 5th to 16th week after immunization.Specific anti- Fusarium IgY protein was prepared by salting out method using ammonium sulfate.The obtained protein solution was put into a freeze dryer and made into freeze-dried powder stored at 4 ℃.The hens in the negative control group were injected with 0.9% sodium chloride to prepare the non-specific antibody as the negative control.Bradford method was used to determine the concentration of specific egg IgY protein and the indirect enzyme-linked immunosorbent assay (ELISA) was employed to measure its titer.The 1×10 5 CFU/ml and 1×10 3 CFU/ml Fusarium suspension were cultured with different concentrations of IgY and phosphate buffered saline (PBS) for 4 days, and the absorbance value at 600 nm was measured.The co-incubated PBS/negative IgY with Fusarium solution was set as blank control/negative control accordingly.The concentration-killing curve of anti- Fusarium IgY against Fusarium was obtained.The specific IgY solution was diluted to 0.02 mg/ml with PBS pH 7.4, and the diluted specific IgY solution was placed into the water bath for 30 minutes at 30, 40, 50, 60, 70, 80, 90 ℃, respectively, and was cooled down to room temperature.The specific IgY solution was diluted to 0.02 mg/ml with PBS pH 1, pH 2, pH 3, pH 4, pH 5, pH 6, pH 7, pH 8, pH 9, pH 10, pH 11, pH 12, respectively, and the diluted specific IgY solution was placed at 4 ℃ for one hour.The activity of diluted specific IgY solution by different methods was measured by indirect ELISA, and the tolerance of IgY to various temperatures and pH was evaluated.Twelve 8-week-old SPF female C57BL/6 mice were selected and randomized into the PBS control group and specific IgY treatment group according to the random table method, with 6 mice in each group.The right eyes of the 12 mice were infected with Fusarium to establish mice model of fungal keratitis.One day after modeling, 200 mg/ml of anti- Fusarium IgY was dropped to the right eyes of mice in the specific IgY treatment group, and PBS was dropped to the right eyes of mice in the PBS control group.The corneas of mice in the two groups were observed under the slit lamp microscope at 1, 3 and 5 days following modeling, and the corneal ulcer was scored according to the grading scale for inflammation score.The use and care of experimental animals followed the Association for Research in Vision and Ophthalmology statement.This study protocol was approved by an Ethics Committee of The Affiliated Hospital of Qingdao University (No.QYFYWZLL26168). Results:The IgY protein concentration from the 5th to 16th week after immunization was 1.57, 2.89, 24.98, 25.09, 23.89, 25.78, 21.57, 21.37, 18.98, 15.78, 14.67, 12.67 mg/ml, respectively.The titer of IgY was increased from the 5th week, and it reached the highest titer 1∶10 000 at the 7th week, which could be maintained until the 12th week after immunization before it dropped gradually.The concentration-killing curve showed that compared with the blank control group and negative control group, Fusarium grew slowly in the specific IgY treatment group.The specific IgY with a titer greater than 1∶10 000 had thermal stability below 60 ℃.The activity of specific IgY was highest at pH 4 to 6, which could be maintained above 70% at pH 3 to 9 and was further reduced with the decrease or increase of pH.At 1, 3 and 5 days after Fusarium infection, the inflammation scores were 3.50±0.55, 7.33±0.82, 4.00±0.63 in the PBS control group, and 3.33±0.82, 4.17±0.75, 2.50±0.55 in the specific IgY treatment group.There was a statistically significant overall difference in inflammation scores at various time points between the two groups ( Fgroup=247.35, P<0.05; Ftime=23.19, P<0.05). At 3 and 5 days after Fusarium infection, there was a smaller ulcer area and decreased inflammation scores in the specific IgY treatment group compared with the PBS control group, and the differences were statistically significant (all at P<0.05). Conclusions:The high titer specific IgY can be successfully prepared by salting out method using ammonium sulfate, which is with high stability, tolerance to temperature and pH.Moreover, it can alleviate the severity of corneal ulcers and reduce inflammation scores in the mouse model of fungal keratitis.