Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

Phage therapy is one of the most important tools for the treatment of infections with multi-drug resistant bacteria. Such phages are usually isolated from hospital effluents, however, no systematic study on the distribution of phages in hospital effluents has been conducted so far. The aim of this study was to isolate the corresponding phages of common pathogenic bacteria isolated in the clinic as hosts, so as to assess the ecological distribution of phages in hospital wastewater and to provide a reference for the isolation and application of phages of drug-resistant bacteria in the clinic. A cross-sectional study design was used in this study. The 125 pathogenic bacteria (belonging to 16 different strains) isolated from the clinical microbiology laboratory of Qilu Hospital of Shandong University from May to June 2023 were selected as the target strains, and the phages corresponding to these strains were isolated and purified from the hospital wastewater by using the double-layer plate sandwich method. At the same time, the distribution of pathogenic bacteria in the same batch of wastewater was analyzed with the help of mNGS sequencing technology, so as to preliminarily investigate the abundance correspondence between pathogenic bacteria and phages in wastewater. The results showed that a total of 56 phage strains were isolated from 125 clinical pathogens as hosts, corresponding to six pathogens, including Acinetobacter baumannii, Klebsiella pneumoniae, Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Stenotrophomonas maltophilia. All six pathogenic bacteria contained strains with different degrees of drug resistance, with a higher percentage of multi-drug resistant strains in A. baumannii, Escherichia coli and P. aeruginosa. The phage acquisition rates of these six pathogens were, in descending order, Escherichia coli (80%), Stenotrophomonas maltophilia (75%), Pseudomonas aeruginosa (70%), Klebsiella pneumoniae (66.67%), Acinetobacter baumannii (36.36%) and Staphylococcus aureus (12.5%). Preliminary mNGS sequencing results showed that the pathogenic bacteria with higher abundance in the batch of effluent were Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Enterococcus faecalis, Acinetobacter baumannii, Klebsiella aerogenes, Klebsiella michiganensis and Pseudomonas aeruginosa. In conclusion, phages of most common clinical Gram-negative pathogens were isolated from hospital wastewater with high isolation rates; however, phages of Gram-positive pathogens were isolated at lower rates, and only phages corresponding to Staphylococcus aureus were isolated in this study. The corresponding mNGS sequencing results showed that the distribution of Gram-negative pathogens in sewage may had a positive correlation with the ecological distribution of phages.

ObjectiveTo explore the effect and possible mechanism of electroacupuncture （EA） at Fenglong （ST40） on liver lipid synthesis and insulin resistance （IR） in hyperlipidemic （HLP） rats. MethodEighteen rats were randomly divided into three groups， blank group， model group， and EA group， each consisting of six rats. The blank group rats were with fed a basic diet， while those in the model group and EA group were fed high-fat diet for 8 weeks. After modeling， the rats in the EA group received bilateral EA treatment at “Fenglong” （ST 40）. The rats in the model group underwent daily binding treatment， once a day， continuously 5 days a week， for a total of 4 weeks. Following the intervention， the levels of triglycerides （TG） and free fatty acids （FFA） in liver tissue was determined using ELISA. Serum TG， FFA， fasting insulin （FINS）， alanine transaminase （ALT）， aspartate Transaminase （AST）， tumor necrosis factor-ɑ （TNF-ɑ）and interleukin 6 （IL-6） were also measured. The fasting plasma glucose （FBG） assessed using a glucose meter and the homeostatic model assessment of insulin resistance （HOMA-IR） was calculated. Liver pathology was examined through HE staining and oil red O staining. The expression of hepatic sterol regulator binding protein 1c （SREBP1c）， recombinant fatty acid synthase （FASN） and stearoyl-CoA desaturase 1 （SCD1） were detected through immunofluorescence. The protein expression levels of liver insulin receptor substrate 1 （IRS1） and tyrosine-phosphorylated insulin receptor substrate 1 （p-IRS1-Tyr） were determined via Western blot. ResultsWhen compared to the blank group， the model group of rats exhibited elevated serum and liver tissue levels of TG and FFA， as well as increased serum levels of AST， ALT， TNF-α， IL-6， FBG， FINS， and HOMA-IR （P＜0.05）. HE staining revealed disordered arrangements of liver cells， indicating widespread fatty degeneration. Oil red O staining showed abundant bright red lipid droplets within liver cell cytoplasm， indicating severe lipid accumulation. The average fluorescence intensity of SREBP1c， FASN， and SCD1 in liver tissue significantly increased （P＜0.05）， while p-IRS1-Tyr protein expression levels significantly decreased （P＜0.05）. In comparison to the model group， the EA group of rats showed significantly reduced serum and liver tissue levels of TG and FFA， along with decreased serum levels of AST， ALT， TNF-α， IL-6， FBG， FINS， and HOMA-IR （P＜0.05）. HE staining indicated more regular arrangements of liver cells， and oil red O staining revealed a significant reduction in liver cell lipid droplets， indicating a less severe degree of lipid accumulation. The average fluorescence intensity of SREBP1c， FASN， and SCD1 in liver tissue significantly decreased （P＜0.05）， while p-IRS1-Tyr protein expression levels significantly increased （P＜0.05）， with no significant difference in IRS1 protein expression （P＞0.05）. ConclusionEA at “Fenglong” （ST 40） can significantly decrease serum lipid in HLP rats， improves liver fat accumulation， and also ameliorate insulin resistance. The mechanism may be related to the inhibition of hepatic lipid synthesis molecule expression， reduced serum inflammatory factors， and an increase in insulin substrate receptor phosphorylation levels.

Objective:To explore any effect of electroacupuncture (EA) at the Zusanli point on the scorch death of duodenal cells in rats with functional dyspepsia (FD) and possible mechanisms.Methods:Twenty-four 7-day-old Sprague-Dawley rats were randomly divided into a blank group, a model group and an EA group, each of 8. FD was induced in both the model and EA group rats using iodoacetamide gavage with tail-clip stimulation. After successful modeling the EA group was given acupuncture at the Zusanli point and then connected with a Korean acupuncture point nerve stimulator for 2 weeks. The other 2 groups were not given any intervention. The rats′ body weight was recorded before and after the modeling, as well as 7 and 14 days later. The gastric emptying rate and the small intestine propulsion rate of the three groups were detected right after the EA intervention, and the serum expression levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured using enzyme-linked immunoassays. Real-time fluorescence quantitative polymerase chain reactions were used to detect the transcription levels of IL-1β and IL-6 in the rats′ duodenums, while western blotting was employed to assess the expression of caspase-1 P20 and dermatin D (GSDMD) in their duodenums.Results:After successful modeling, the average body weight of the rats in the model and EA groups was significantly different from that in blank group, and after 7 and 14 days the average body weight of the former groups was significantly different from that of the blank group, with significant differences between the two groups as well. After the EA intervention significant differences were observed in gastric reside and small intestine propulsion rate between the EA group and the model group, as well as between the model and the blank group. After the intervention, there were significant differences between the blank group and the other two groups in the average expression of IL-1β and IL-6 in serum, IL-1β and IL-6 mRNA in the duodenum, as well as the GSDMD and caspase-1 p20 proteins in the duodenum. There were significant differences between the model and EA groups in all of the above measurements.Conclusions:EA at the Zusanli point can significantly reduce the level of scorch death in the duodenum of FD rats, as well as relieve low-grade duodenal inflammation and the clinical symptoms of FD. Its mechanism may be related to the down-regulation of the expression of caspase-1 P20 and GSDMD-N protein, and of inflammatory factors such as IL-1β and IL-6, relieving low-grade duodenal inflammation.

Objective:To explore the effect of tibial nerve vibration on triceps surae spasticity in stroke survivors and its electrophysiological mechanism.Methods:Thirty stroke survivors with upper limb spasticity were randomly divided into a treatment group and a control group, each of 30. Both groups were given routine rehabilitation training while the treatment group was additionally provided with local vibration of the tibial nerve with an amplitude of 0.3mm at 60Hz. Before and after the treatment, the modified Ashworth scale (MAS) and Clonus grading were used to assess muscle tone and spasticity. Electrophysiological functions were evaluated using the tibial nerve motor and sensory conduction test and F wave and H reflex sensing.Results:Before the treatment there were no significant differences between the two groups in their average MAS scores, Clonus grading, maximum H amplitude (Hmax), the ratio of maximum H to maximum M amplitude (H/M) or the intensity of stimulus required to elicit Hmax. After the treatment, however, all of those measurements had improved significantly in both groups with the average improvements in the treatment group significantly greater than those in the control group.Conclusions:Local vibration of the tibial nerve combined with traditional rehabilitation is more effective than traditional rehabilitation alone in relieving triceps surae spasticity and reducing muscle tone after a stroke. The vibration seems to inhibit excitation of the reflex pathway.

Objective : To analyze the effect of different cavosurface angles on the stress distribution of ClassⅠ cavity composite resin filling of molars through the three-dimensional finite element method and to provide references for the preparation of ClassⅠ cavities. Methods: Three-dimensional finite element models of ClassⅠ composite resin filling of mandibular first molars with three different cavosurface angles (group A: 90°, group B: 120°, group C: 135°) were established. Polymerization shrinkage of composites was simulated with a thermal expansion approach. The mechanical behavior of the restored models in terms of stress and displacement distributions under the combined effects of polymerization shrinkage and occlusal load (600 N) was analyzed. Results: For ClassⅠ cavities with the same cavity size, the total stress of the restoration model and the maximum stress of the enamel in group A were less than those in groups B and C after cavity composite resin restoration with three cavity cavosurface angles (in which the width of the enamel bevel was 1 mm in groups B and C). The maximum stress of the dentin and adhesive was similar in the three groups, the maximum stress of the composite in group C was the largest, and the maximum stress of the composite in group B was the smallest. In terms of stress distribution, the maximum stress in each restoration model was mainly concentrated in the enamel at the cavosurface, near the enamel-dentin interface and at the edge of the restoration material. Conclusion From the point of reducing the stress of residual tooth tissue, the preparation of 90° angle without enamel bevel is an ideal method for cavity preparation when composite resin is used to fill ClassⅠ cavities of molars.

Objective:To realize prokaryotic expression, purification and identification of 2019-novel Coronavirus (2019-nCoV) nucleocapsid protein (NP), and apply it to the serological diagnosis.Methods:The synthetic 2019-nCoV NP gene was cloned into the prokaryotic expression vector pET28a to construct expression plasmid, and then purified by Ni-chelating affinity. SDS-polyacrylamide gel electrophoresis (SDS-PAGE), indirect enzyme-linked immunosorbent assay (ELISA), Western blot (WB), and immunochromatography were used to test the purified protein. Indirect ELISA reaction conditions were optimized for serum antibody detection.Results:The relative molecular mass of recombinant NP was about 50×10 3 after SDS-PAGE electrophoresis, which was consistent with the expectation. Indirect ELISA and WB results showed that it could specifically bind to the serum of patients infected with 2019-nCoV. The detection limit of NP was 0.2 ng/ml by immunochromatography. The sera from 32 patients infected with 2019-nCoV and the control sera were detected by indirect ELISA, and the results showed that they were clearly clustered. Conclusions:Prokaryotic expression of 2019-nCoV NP has good immunogenicity and can be used for the development of serological diagnostic reagents.

Objective:To analyze the effects of protein intake of varied dietaries on the pregnancy of pregnant women.Methods:The pregnant women who had their medical records in our hospital were included in this study.Their dietary were collected based on the respective review of their food during 24 hours when they visited the Nutrition Clinics between 24 weeks and 28 weeks.The subjects were divided into six groups according to the dietary calories and protein intake:group A (normal calorie and normal protein),group B (normal calorie and low protein),group C (normal calorie and high protein),group D (high calorie and normal protein),group E (high calorie and high protein) and group F (low calorie and low protein).The gestational diabetes mellitus incidence rates,levels of fasting blood glucose (FBG),blood glucose (BG),hemoglobin Alc (HbAlc),and blood lipids,and bone mineral density were compared among six groups.Results:The gestational diabetes mellitus incidence rate,levels of postprandial blood glucose at 1 hour and 2 hour in groups of B,C,D,and E were significantly higher than those in group A (P ＜ 0.05).The levels of triglyceride,total cholesterol,and low density lipoprotein cholesterol were remarkably higher in groups of B,D,and E than those in group A (P ＜ 0.05).The bone mineral density was significantly lower in group A than that in groups of C,E,and F (P ＜ 0.05).Conclusion:The diet with normal calorie and normal protein is the scientific dietary structure.The pattern with high calorie and high protein intake is not recommended for pregnant women.

OBJECTIVETo compare the clinical therapeutic effects and safety on menopausal symptoms between manual acupuncture (MA) and electroacupuncture (EA).

METHODSFifty patients were randomized into an MA group (25 cases) and an EA group (25 cases). In the MA group, the regular needling technique and pseudo-EA were used at Guanyuan (CV 4), Zigong (EX-CA 1), Tianshu (ST 25) and Sanyinjiao (SP 6). In the EA group, the acupoints were the same as the MA group and stimulated with EA and pseudo-MA (no manipulation applied in treatment), with disperse-dense wave, 10 Hz/50 Hz, 0.5 to 1.0 mA. In the two groups, the needles were retained for 30 min. The treatment was given once every two days, three times a week, totally for 8 consecutive weeks. The results of the menopause rating scale (MRS), the menopause-specific quality of life (MENQOL), the self-rating anxiety scale (SAS) and the self-rating depression scale (SDS), follicle stimulating hormone (FSH), luteinizing hormone (LH) and estradiol (E) as well as adverse reactions were evaluated before treatment, in 4 and 8 weeks of treatment separately.

RESULTSCompared with those before treatment, the scores of MRS, MENQOL, SAS and SDS were all reduced in the 4 and 8 weeks of treatment in the two groups (all<0.05). The results in 8 weeks of treatment were lower than those in 4 weeks of treatment in the two groups (all<0.05). The differen-ces were not significant statistically between the two groups (all>0.05). Compared with those before treatment, the levels of FSH and LH reduced and Eincreased after treatment in the two groups, without significant differences (all>0.05). The differences were not significant statistically between the two groups (all>0.05). The severe adverse reactions were not found in the treatment of the two groups.

CONCLUSIONSBoth manual acupuncture and electroacupuncture relieve the symptoms of depression and anxiety in menopausal syndrome,improve the living quality and do not induce apparent changes in serological sex hormones in the patients.

Objective To introduce our experiences in ear reconstruction with Ba Da Chu Method during the past 10 years, and to summarize the operative skills and key points .Methods Generally, Ba Da Chu Method for ear reconstruction includes 3 stages:Stage 1, skin expansion in mastoid region .Stage 2, auricular reconstruction), consisting of tissue expander removal, earlobe transposition, expanded skin flap and temporal fascia flap formation , autologous costal cartilage harvest , framework fabrication and transplantation , and the wound closure .Stage 3: tragus formation , conchoplasty , and refinement of the reconstructed ear.Results From January, 2006 to December, 2015, we performed 5628 reconstructed ears for 5267 patients with congenital microtia .Follow-up period ranged from 1 to 10 years.Complications in stage 1 included hematoma , infection and expander exposure sporadically arising , which were treated properly and stage2 operations were carried out on time or delayed .5202 (92.4％) reconstructed ears were demonstrated with fine substructure landmarks .236(4.2％) reconstructed ears with poor blood circulation at the margin of skin flap , were cured completely and ended up with acceptable outcomes;108 ( 1.9％) reconstructed ears , manifestedwith mild cartilage framework exposure due to partial necrosis of skin graft , were repaired in stage 3.61 (1.1％) of them with severe exposure , needed additional operations to cover the framework with axial fascial flap immediately .21 ( 0.3％) reconstructed ears lost the normal contour because of cartilage infection , which a secondary operation was needed to repair .The complications of stage 3 occurred rarely , which could be cured in the end .Conclusions Ba Da Chu Method is well adapted to treat congenital microtia , and it is flexible to adjust surgical skills when microtia was combined with other complicated soft tissue deformities or craniofacial bone defects;Satisfactory result were achieved in 92.4％reconstructed ears in this study with fine substructures;During approximate 2-month skin expansion , great attention must be paid to ensure successful expansion , which is based on extensive clinical experiences from doctors, appropriate nursing management from nurses , and careful observation from patients or their families.