Objective To investigate the expression of neuroprotective peptide [Gly14]-Humanin (HNG) in eukaryotic cells by gene engineering technique and analyze its biological activity. Methods By means of asymmetrical primer/template, double stranded cDNA of HNG with FLAG in its C-terminal was obtained, which was cloned into the plasmid pcDNA3.1(-), and the resultant recombinant vector pcDNA3.1(-)/HNG-FLAG was transfected into PC12 cells. At the same time, the recombinant vector pcDNA3.1(-)/EGFP was transfected to control the efficiency of transfection. The expression of HNG in the cells was determined by immunocytochemistry. In order to analyze the biological activity of the expressed HNG, 25μM Aβ25-35 peptide was added to the culture medium of the transfected cells for 24h, then cell morphology, MTT assay and Hoechst 33258 staining were observed. Results The eukaryotic expression vector of pcDNA3.1(-)/HNG-FLAG was identified by enzyme digestion and sequencing. HNG was highly expressed in PC12 cells. After exposure of PC12 cells to 25μM Aβ25-35 for 24h, cell viability decreased to (65.8±5.3)%, and the dystrophic changes of neuritis and nuclei condensation were obvious. When cells were pre-transfected with pcDNA3.1(-)/HNG-FLAG, Aβ25-35-induced cell death and morphological changes of cells and nuclei were suppressed. In contrast, pre-transfected with empty vector did not protect cells from Aβ25-35-induced toxicity. Conclusion The eukaryotic expression vector for FLAG-tagged HNG was successfully constructed and expressed in PC12 cells. Expressed HNG has biological activity.

Objective To discuss the effect of rehabilitation nursing for total knee arthroplasty by discussing hospitalization expenses and its factors of rehabilitation nursing.Methods 305 patients who were underwent total knee arthroplasty from 2011 to 2013 were recruited and recovered according to rehabilitation nursing and training requirements established by our hospital.Patients admitted in 2011 were designated as control group.The comparison of American knee society knee score and knee rang of motion of patients were made and analyzed.Results The annual comparison of sores of pre-and post-operative KSS (t=5.30,7.99,11.20),and knee ROM of patients (t=8.21,4.57,7.86) showed that the difference was statistically significant (P＜0.05).The total costs and rehabilitation nursing expenses inreased year by year (P＜0.05).The proportion of rehabilitation and rehabilitation nursing expenses each year were 5.77％ (3 157/54 679),4.60％ (2 847/61 831),5.15％ (3 341/64 930) and 0.51％ (280/54 679),0.64％ (393/61 831),0.52 ％(338/64 930),respectively.Conclusions Pre-and post-operative correct rehabilitation exercises are one of the most important guarantee of curative effect.There is a lower percentage of rehabilitation nursing expenses in total costs,and the value of the nursing staff has not been fully reflected.

Objective To explore the effectiveness and adverse effect of high frequency oscillation ventilation (HFOV) combined with Sildenafil (SIL) treatment on newborns with persistent pulmonary hypertension (PPHN).Methods A total of 89 cases of PPHN infants collected from Chengdu Women and Children's Central Hospital from Sep.2010 to Sep.2012 were randomly divided into HFOV group,constant mechanical ventilation (CMV) group,HFOV combined SIL group (HFOV + SIL group) and CMV combined with SIL group (CMV + SIL group).The arterial blood gas,pulmonary artery pressure (PAP) and adverse reactions were monitored before and 3 days after treatment.SNK multiple comparison method andx2 test were performed for data before and after treatment among groups for continuous variables and categorical variables,respectively.Results The levels of pa (O2) [(79.1 ± 13.7) mmHg (1 mmHg =0.133 kPa),(77.9 ±14.6) mmHg,(85.4 ±15.2) mmHg],Sa(O2) [(87.8 ±13.4)％,(88.4±15.6)％,(96.1±15.9)％],pa(CO2)[(42.5±11.3) mmHg,(40.2 ±10.5) mmHg,(35.6 ±8.7) mmHg] and PAP [(31.1 ± 8.1) mmHg,(30.4 ± 9.5) mmHg,(25.8 ± 7.3) mmHg] were all improved significantly in CMV + SIL group,HFOV group and HFOV + SIL group compared with those in CMV group[(69.9 ± 12.3) mmHg,(81.1 ± 14.9)％,(48.1 ±9.5) mmHg,(35.6 ±8.9) mmHg] (F =4.629 3,3.673 2,5.865 3,4.849 5,P ＜0.05),especially for HFOV + SIL group(P ＜ 0.05).No significant difference in such indicators was observed between CMV + SIL group and HFOV group (P ＞ 0.05).The effective rate in HFOV + SIL group (90％) was the highest among the 4 groups (x2 =7.938,P ＜ 0.05).During the treatment,all neonates have no adverse reaction.Conclusion The combined use of SIL and HFOV might be a more effective and safer method in the treatment of PPHN of neonate.

Objective To assess the differences in nutrient composition by chemical analysis method and weighing method,A corps of armed police forces daily dietary nutrient intakes were investigated in Tianjin,to understand the status of the actual intakes of dietary nutrient.Methods A dietary survey was performed using the weighing method and with ref-erence to China food composition database in 2002.The intake of each nutrient was calculated through chemical analysis;Nutritional analysis was used in the collection of duplicate food portions during the dietary survey.Results For most of the main dietary nutrients(protein,fat and carbohydrates),minerals (Na,K,Mg,Ca,Zn,Cu and Se)and vitamins (β-car-otene,vitamin E,thiamine,riboflavin,vitamin C,niacin,and vitamin A),the calculated values of intakes were found to exceed those obtained via chemical analysis.Conclusion Differences of the two methods of dietary survey were ultimately due to the analysed values were below the calculated ones.So it is necessary to improve methods of food storage and trans-portation in order to obtain adequate nutrition and to improve the health of troops.

BACKGROUNDTo investigate the immunohistochemical detected method and the clinical incidence of the bone marrow micrometastases (BMM) in patients with non-small cell lung cancer (NSCLC) and to analyze the sensitivity and specificity and clinical application value.

METHODSBone marrow samples were collected from the anterior superior iliac spines or posterior superior iliac spines of 53 patients with NSCLC in clinical stage I to III and 15 patients in stage IV, and the BMM was detected by immunohistochemical techniques (IHC) using monoclonal antibodies AE1/AE3 against cytokeratin. Chi-square test was used statistically.

RESULTSThe IHC sensitivity could be 10⁻⁵. The BMM positive rate was 22.6% (12/53) in stage I to III and 53.3% (8/15) in stage IV, and there was a significant difference in the BMM positive rate between stage I to III and stage IV (P < 0.05). No correlation was observed between BMM and sex, age, KPS, pathology classification and cancer cell differentiation.

CONCLUSIONSThe detection of BMM by IHC is convenient, sensitive, and specific. It might be helpful to diagnose bone marrow micrometastasis in patients with NSCLC.

Objective To evaluate the effectiveness and safety of quadruple regimen in preventing multi-day cisplatin-based chemotherapy-induced nausea and vomiting(CINV).Methods A total of 112 patients with malignant tumors who underwent cisplatin-based chemotherapy.According to the random number table method,patients were divided into the experimental group and the control group,with 56 cases in each group.The control group received cisplatin-based chemotherapy and triple therapy of fosapreitant dimeglumine,ondansetron hydrochloride and dexamethasone tablets.On this basis,the experimental group was given quadruple therapy containing olanzapine tablets.The occurrence of nausea and vomiting,as well as changes in FLIE and HAD scores were observed in both groups.Results The incidence of nausea and vomiting was lower from day 1(D1)to day 9(D9)after the beginning of chemotherapy in the experimental group than that in the control group(P＜0.05).The incidence of nausea and vomiting in the experimental group was lower than that in the control group.On D9 after the beginning of chemotherapy,the nausea score,vomiting score and total score of FILE were higher in the experimental group than those in the control group(P＜0.05).There were no significant differences in depression or anxiety score and incidence of adverse reactions on D1 and D9 after the beginning of chemotherapy between the two groups(P＞0.05).Conclusion The quadruple antiemetic regimen can improve the control rate of CINV induced by multi-day cisplatin-based chemotherapy,especially for the control of delayed nausea and vomiting,and improve the quality of life of patients during chemotherapy,with good safety.

Objective To investigate the effect of Yiqi Huoxue Tongluo Decoction on microRNA-126a-5p(miR-126a-5p)and vascular endothelial growth factor(VEGF)signaling pathway in cervical spondylotic myelopathy model rats.Methods Thirty healthy male SD rats were divided into the sham operation group,the model group and the traditional Chinese medicine(TCM)group by random number table method.Cervical spondylotic myelopathy models were prepared in the model group and the TCM group.The TCM group was given intragastric administration of Yiqi Huoxue Tongluo Decoction,while the sham operation group and the model group were given intragastric administration of normal saline for 12 weeks.After intervention,the threshold of mechanical stimulation and retraction time of thermal stimulation in each group were measured by behavior tests.Rats were sacrificed to collect intervertebral disc tissue for hematoxylin-eosin(HE)staining and observe the number of vascular buds in intervertebral disc.Rat intervertebral disc annulus fibrosus cells were subjected to terminal dexynucleotidyl transferase-mediated dUTP nick end labeling(TUNEL)staining.The miR-126a-5p and VEGF mRNA of rat intervertebral disc tissue were detected by real-time fluorescence quantitative polymerase chain reaction(RT-PCR).The expression of VEGF protein of rat intervertebral disc tissue was detected by Western blot assay.Results Compared with the sham operation group,the number of vascular buds in intervertebral disc was decreased in the model group and the TCM group.The cell destruction of intervertebral disc annulus was obvious in rats,and apoptosis was high and cell density decreased.Mechanical stimulation threshold decreased,and mechanical stimulation threshold decreased.The level of miR-126a-5p was decreased,and the expression levels of VEGF mRNA and protein were increased.Compared with the model group,the number of vascular buds in intervertebral disc was increased in the TCM group.The destruction of intervertebral disc annulus cells was alleviated in rats.The apoptosis of annulus fibrosus cells in intervertebral disc decreased and cell density increased.The threshold of mechanical stimulation increased,and the retraction time of thermal stimulation was prolonged.The level of miR-126a-5p increased,and the expression levels of VEGF mRNA and protein decreased(P＜0.05).Conclusion The mechanism of Yiqi Huoxue Tongluo Decoction in the treatment of cervical spondylotic myelopathy may be related to the up-regulation of miR-126a-5p expression and the down-regulation of VEGF expression.

Objective To investigate the effect of semen bacterial infection on semen parameters in male infertility patients and analyze of the drug resistance profile of the bacterial isolates. Methods According to the results of bacterial culture, the semen samples of 74376 infertile men collected between April, 2016 and April, 2017 were divided into infection group and non-infection group, and the infertility type and semen parameters were compared between the two groups. The bacterial species and the drug sensitivity of the isolates were analyzed. Results Bacterial infections were detected in 1.38％of the total semen samples collected. The positivity rate of semen bacterial infection was 1.41％in normal semen group, 1.55％in asthenospermia group, 1.18％ in oligospermia group, 1.57％ in asthenospermia/ oligospermia group, and 0.17％ in azoospermia group. The positivity rate was lower in azoospermia group than in the other groups. Bacterial infection mainly affected sperm motility (P<0.05), sperm density and forward motile sperm ratio (P<0.01). The most common bacterial species causing the infections included, in the descending order of frequencies, Escherichia coli (63.59％), Klebsiella pneumoniae subspecies (19.80％) and Proteus mirabilis (13.22％). Drug susceptibility tests showed that the isolates of Escherichiacoli, Pneumonia Klebsiella pneumonia subspecies and Citrobacter koseri were commonly resistant to amoxicillin; Pseudomonas aeruginosa isolates were resistant to Bactrim and ampicillin/sulbactam; Staphylococcus aureus isolates were resistant to penicillin. Conclusion Male infertile patients have a low bacterial infection rate in the semen. Bacterial infection severely affects the sperm fertilization process by causing impairment of sperm motility and lowered sperm density. Escherichia coli is the most common pathogenic bacterium for the semen infection. With the exception of Staphylococcus aureus, the other bacterial strains isolated were found to be sensitive to imipenem and meropenem.

Objective:To evaluate the upper airway dimension changes in mandibular prognathism accompanied with maxillary retrognathism patients treated with bimaxillary orthognathic surgery by spiral CT.Methods:The data of patients with mandibular prognathism accompanied with maxillary retrognathism who underwent bimaxillary surgery in the Department of Oral and Maxillofacial Surgery of Nanjing Stomatological Hospital, Medical School of Nanjing University from May 2017 to October 2020 were retrospectively analyzed. The surgical method was LeFort Ⅰ osteotomy and bilateral sagittal split mandibular osteotomy. The patients were divided into 2 groups based on the sagittal setback distance of the supramentale(in group A, the setback distance of the supramentale was more than 5 mm; in group B, the setback distance of the supramentale was less than or equal to 5 mm). Spiral CT data were collected 1 week before surgery(T0), 1 month after surgery(T1), and 6-12 months after surgery(T2). Three-dimensional model reconstruction was performed. The airway cross-sectional measurements, length and volumes in T0, T1 and T2 were measured. The effect of bimaxillary orthognathic surgery on upper airway of the patients was evaluated quantitatively. We performed repeated measures analysis of variance to compare the differences of upper airway among T0, T1 and T2 in the same group. The Bonferroni method was used for multiple comparisons if the difference was statistically significant(α=0.017). We used two-factor repeated measures analysis of variance to compare the differences of the upper airway change trend between the two groups.Results:A total of 30 patients were included, including 15 patients in group A, 5 males and 10 females[aged: (21.2±2.3) years]; and 15 patients in group B, 7 males and 8 females[aged: (23.6±2.4) years]. The cross-section area and sagittal diameter of lower velopharyngeal plane(Lvp), the glossopharynx airway volume and total upper airway volume: group A decreased significantly at T1, and did not return to T0 level at T2(all P<0.017); group B decreased significantly at T1(all P<0.017), and returned to T0 level at T2(all P>0.017); the change trend of these indexes between the two groups was statistically significant(all P<0.05). The airway length of the glossopharynx and the total upper airway: group A increased significantly at T1 and T2(both P<0.017); group B had no significant changes at T1 or T2(both P>0.017); the change trend of these two indexes between the two groups was statistically significant(both P<0.05). Conclusions:The glossopharynx airway volume and total upper airway volume decreased in mandibular prognathism accompanied with maxillary retrognathism patients after bimaxillary orthognathic surgery. If the setback distance of the mandible is less than or equal to 5 mm, they can probably return to the preoperative level 6-12 months after surgery. If the setback distance of the mandible is more than 5 mm, they can not return to the preoperative level 6-12 months after surgery.

Objective:To evaluate the upper airway dimension changes in mandibular prognathism accompanied with maxillary retrognathism patients treated with bimaxillary orthognathic surgery by spiral CT.Methods:The data of patients with mandibular prognathism accompanied with maxillary retrognathism who underwent bimaxillary surgery in the Department of Oral and Maxillofacial Surgery of Nanjing Stomatological Hospital, Medical School of Nanjing University from May 2017 to October 2020 were retrospectively analyzed. The surgical method was LeFort Ⅰ osteotomy and bilateral sagittal split mandibular osteotomy. The patients were divided into 2 groups based on the sagittal setback distance of the supramentale(in group A, the setback distance of the supramentale was more than 5 mm; in group B, the setback distance of the supramentale was less than or equal to 5 mm). Spiral CT data were collected 1 week before surgery(T0), 1 month after surgery(T1), and 6-12 months after surgery(T2). Three-dimensional model reconstruction was performed. The airway cross-sectional measurements, length and volumes in T0, T1 and T2 were measured. The effect of bimaxillary orthognathic surgery on upper airway of the patients was evaluated quantitatively. We performed repeated measures analysis of variance to compare the differences of upper airway among T0, T1 and T2 in the same group. The Bonferroni method was used for multiple comparisons if the difference was statistically significant(α=0.017). We used two-factor repeated measures analysis of variance to compare the differences of the upper airway change trend between the two groups.Results:A total of 30 patients were included, including 15 patients in group A, 5 males and 10 females[aged: (21.2±2.3) years]; and 15 patients in group B, 7 males and 8 females[aged: (23.6±2.4) years]. The cross-section area and sagittal diameter of lower velopharyngeal plane(Lvp), the glossopharynx airway volume and total upper airway volume: group A decreased significantly at T1, and did not return to T0 level at T2(all P<0.017); group B decreased significantly at T1(all P<0.017), and returned to T0 level at T2(all P>0.017); the change trend of these indexes between the two groups was statistically significant(all P<0.05). The airway length of the glossopharynx and the total upper airway: group A increased significantly at T1 and T2(both P<0.017); group B had no significant changes at T1 or T2(both P>0.017); the change trend of these two indexes between the two groups was statistically significant(both P<0.05). Conclusions:The glossopharynx airway volume and total upper airway volume decreased in mandibular prognathism accompanied with maxillary retrognathism patients after bimaxillary orthognathic surgery. If the setback distance of the mandible is less than or equal to 5 mm, they can probably return to the preoperative level 6-12 months after surgery. If the setback distance of the mandible is more than 5 mm, they can not return to the preoperative level 6-12 months after surgery.