Objective: To study the relationship between sugar sweetened beverages consumption characteristics and insomnia of college students in Yunnan Province, so as to provide evidence for sleep quality improvement of college students. Methods: A cluster random sampling method was used to select 2 515 college students from two universities (Kunming University and Dali Nursing Vocational College) in Kunming and Dali in Yunnan Province for a longitudinal study, including baseline survey (T1, November 2021) and three follow up surveys (T2: June 2022, T3: November 2022, T4: June 2023). Sugar sweetened beverages consumption of college students was collected by Semi quantitative Food Frequency Questionnaire and insomnia was assessed by Insomnia Severity Index Scale. Sugarsweetened beverages consumption was analyzed by Kruskal-Wallis test. The Mann-Whiter U test and Kruskal-Wallis test were used to compare the detection rate of insomnia in college students with different population characteristics, and the generalized estimating equations model was established to analyze the association between sugar sweetened beverages consumption and insomnia. Results: The reported rate of insomnia among college students from T1 to T4 was 21.2%, 23.6%, 30.5 % and 36.0%, respectively. The median of sugar sweetened beverages consumption per week was 5 (1,9) bottles per person, and there were significant differences in sugar sweetened beverages (carbonated beverages, fruit beverages, tea beverages, milk beverages, energy beverages) consumption among college students with different insomnia status ( χ 2=42.91, 23.67, 29.98, 61.70, 30.82, P <0.01). The analysis of the generalized estimating equation model revealed that the consumption of carbonated beverages ( β= 0.04, 95%CI =0.00-0.08) and the consumption of milk beverages among college students ( β=0.04, 95%CI =0.00-0.09) were correlated with insomnia ( P <0.05). The stratified analysis indicated that consumption of carbonated beverages by female college students was associated with insomnia [ β(95%CI )=0.06(0.01-0.11)]; consumption of milk beverages among college students from middle income family was associated with insomnia [ β (95% CI )=0.05(0.00-0.10)], and consumption of carbonated beverages and fruit beverages from college students with high household economic status were both associated with insomnia [ β (95% CI )=0.35(0.23-0.46), 0.12(0.00-0.24)] ( P <0.05). Conclusion Sugar sweetened beverages, especially carbonated beverages, are associated with insomnia among college students in Yunnan Province.

Objective:To investigate the effect of bedside high-flow continuous blood purification (CBP) combined with Xuebijing in the treatment of severe sepsis (SS) and the influence on the patient′s coagulation-fibrinolysis index, immunity index and expression of peripheral blood Toll-like receptor 4 (TLR4).Methods:Ninety-three patients with SS who were admitted and treated in the Lianyungang First People′s Hospitalfrom January 2017 to October 2019 were selected. They were divided into the combined group (51 cases, treatment with bedside high-flow CBP and Xuebijing injection based on bundle therapy) and the control group (42 cases, treatment with Xuebijing injection based on bundle therapy). The changes in coagulation and fibrinolysis index, immunity index, biochemical index such as TLR4 before treatment and after 1 week of treatment were compared between the two groups. The incidences of complications in both groups were statistically analyzed, and the discharge time from ICU, mechanical ventilation time and 28-day mortality were recorded.Results:After 1 week of treatment, the levels of prothrombin time (PT) and activated partial thromboplastin time (APTT) in the two groups were shortened, D-dimer (D-D) and fibrinogen (FIB) were decreased ( P<0.05); and the levels of PT and APTT in the combined group were shorter than those in the control group, the levels of DD and FIB were lower than those in the control group, there were statistical differences ( P<0.05). After 1 week of treatment, the levels of CD 4+ and CD 4+/CD 8+ ratio in both groups were increased ( P<0.05), and the levels of CD 4+ and CD 4+/CD 8+ ratio in the combined group were higher than those in the control group ( P<0.05). After 1 week of treatment, the levels of TLR4, C-reactive protein (CRP), procalcitonin (PCT), white blood cell count (WBC), blood lactate (Lac), blood urea nitrogen (BUN) and serum creatinine (Scr) in both groups were decreased ( P<0.05), meanwhile, the above indexes in the combined group were lower than those in the control group ( P<0.05). The incidence of multiple organ failure and the 28-day mortality rate in the combined group were lower than those in the control group: 3.92%(2/51) vs. 19.05%(8/42), 13.73%(7/51) vs. 30.95%(13/42), there were statistical differences ( P<0.05). The discharge time from ICU and mechanical ventilation time in the combined group were shorter than those in the control group: (12.35 ± 2.14) d vs. (14.17 ± 3.36) d, (7.12 ± 2.23) d vs. (8.51 ± 2.39) d, there were statistical differences ( P<0.05). Conclusions:Bedside high-flow CBP combined with Xuebijing injection in the treatment of SS can improve the patient′s condition, regulate the balance of coagulation and fibrinolysis, avoide the activation of coagulation, inhibite inflammatory response, reduce the expression of TLR4 in peripheral blood, improve immune function, protecte kidney function and promotethe patient′s recovery.

Objective:To preliminarily evaluate the effect of tension stimulation on the biological activity of and expression of fibrosis marker genes in keloid fibroblasts (KD-Fbs) .Methods:Three patients who were diagnosed with keloids and received surgical treatment were collected from the Department of Dermatology, Tangdu Hospital, the Fourth Military Medical University from January to March 2017. Human KD-Fbs were isolated from resected keloid tissues, and subjected to primary culture. The third- to sixth-passage KD-Fbs were divided into tension group and control group to be cultured in the tension-based chamber and control chamber respectively, and subjected to tension stimulation and normal culture respectively. Cell counting kit-8 (CCK8) assay was performed to assess the proliferative activity of KD-Fbs after 1-, 2-, 3- and 4-day culture, and the scratch assay to evaluate the migratory ability of KD-Fbs after 1- and 2-day culture. After 48-hour treatment, real-time quantitative PCR and Western blot analysis were performed to determine the mRNA and protein expression of fibrosis markers type Ⅰ collagen, fibronectin and α-smooth muscle actin (α-SMA) in KD-Fbs respectively. Two-independent-sample t test was used for comparisons between 2 groups. Results:CCK8 assay showed that the proliferative activity of KD-Fbs was significantly higher in the tension group than in the control group after 1-, 2-, 3- and 4-day culture ( t=3.05, 7.00, 16.65, 15.19, respectively, all P< 0.05) . After 1- and 2-day culture, the scratch assay showed that the migration rate of KD-Fbs was significantly higher in the tension group (48.65%±3.96%, 100.00%, respectively) than in the control group (9.36%±1.14%, 50.35%±4.23%, t=16.53, 20.35, respectively, both P< 0.01) . Real-time quantitative PCR showed that the mRNA expression of type Ⅰ collagen, fibronectin and α-SMA was significantly higher in the tension group (3.04±0.20, 2.16±0.10, 3.76±0.24, respectively) than in the control group (1.00; t=17.57, 21.01, 20.25, respectively, all P< 0.01) . As Western blot analysis revealed, changes in the protein expression of the 3 fibrosis markers were consistent with their mRNA expression changes (all P< 0.05) . Conclusion:Tension may participate in the fibrosis in keloids by promoting the expression of fibrosis marker genes, and enhancing the proliferative and migratory ability of KD-Fbs.

Objective:To investigate the application of anterolateral thigh chimeric perforator flap in repairing complex soft tissue defects in lower extremities.Methods:From January, 2015 to June, 2019, 76 cases of complex soft tissue defects in lower extremities were repaired with free anterolateral thigh chimeric perforator flap pedicled with the descending branch of lateral circumflex femoral artery, including 29 cases in shank and 47 cases in ankle, and all had various tissue necrosis, infection, deep tissue defect and orthopaedic implant exposure. The size of the wound ranged from 15 cm×8 cm to 35 cm×20 cm, in which 45 cases associated with dead cavity formation, 62 cases with combined fractures at the same site, and 38 cases with combined fractures or other system injuries in other sites. After debridement, VSD treatment, good wound granulation and infection control, the chimeric perforator flap pedicled with the descending branch of the circumflex lateral femoral artery was designed and harvested. The perforator flap was used to repair most of the wounds with deep tissue exposures, the muscle flap was used to fill the dead cavity and (or) cover the wound around the flap, and free skin was grafted of the muscle flap at the first or second stage. Scheduled follow-up was conducted after the operation.Results:All 76 flaps survived, including vascular crisis occurred in 2 cases within 72 hours after surgery and ceased after immediate surgical exploration. One case had a further surgical operation due to excessive bleeding. Partial necrosis occurred at the distal end of the flap or the skin graft area of the muscle flap in 4 cases, of which 2 cases were treated with grafted skin again and the other 2 cases completely eliminated the wound after active dressing change. Other 16 cases of post-traumatic osteomyelitis with bone defect were repaired respectively with bone grafting, bone transportation or Masquelet technique in 3 to 6 months after wound healing. Among the 76 cases, 68 cases had primary wound healing while 8 cases delayed. Seventy-one cases were followed-up from 9 to 24 months, with an average of 16 months, while 5 cases lost. The appearance and function of the affected limbs recovered satisfactorily without recurrence of infection.Conclusion:With the anterolateral thigh chimeric perforator flap transplantation, the perforators flap can be conveniently used to repair the wound, and the muscle flap can be used to fill dead cavity and(or) deep wounds with free skin graft. Only the descending branch of lateral circumflex femoral artery is required to be anastomosed. It achieves a 3-dimensional effective reconstruction of the complex wound in extremities. It is a safe and effective technique to repair and reconstruct the complex wound in lower extremities and ideal in clinical applications.

Brain-derived neurotrophic factor (BDNF) exerts its survival-promoting effects on photoreceptors and retinal ganglion cells, however, delivery systems with little-to-no side effect are needed to sustain its controlled release and long-term efficacy. Our previous studies demonstrated that adipose-derived stem cells (ADSCs) are ideal delivery systems for gene therapy; moreover, ADSCs present unique properties like migration to damaged tissue sites, immunomodulation and anti-inflammation. Herein, we propose to employ ADSCs as the BDNF gene delivery vehicle. Different Analyses like flow cytometry, differentiation and cell proliferation assays etc demonstrated that BDNF were successfully transduced into ADSCs and the stemness of ADSCs was maintained even with the transduction. Real Time PCR and Western blot were used to measure mRNA and protein expressions of the BDNF-transduced ADSCs. The results demonstrated that the BDNF expression level of the lentiviral-BDNF transduced ADSCs is significantly increased and, moreover, enhanced the expression of other neurotrophic and downstream signaling factors. The data suggest that ADSCs are a reliable delivery vehicle for BDNF and could be used for the treatment of various diseases.

Objective: To construct ¹³¹I-the fifth generation polyamidoamine (PAMAM(G5.0)) with targeting peptide Ser-Arg-Glu-Ser-Pro-His-Pro (SRESPHP; SR) or Gly-Pro-Leu-Pro-Leu-Arg (GPLPLR; GP) and double targeting peptide SR/GP, and evaluate the targeting ability in medullary thyroid carcinoma (MTC) model. Methods: PAMAM(G5.0), PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP were radiolabeled with ¹³¹I by chloramine T method. The radiolabeled yield and radiochemical purity were determined by thin layer chromatography. MTC xenografts were developed and the percentage radio-activity of injection dose per gram of tissue (%ID/g) in tumor and organs was measured at 24 h post-injection. Region of interest (ROI) was drawn and the tumor/non-tumor (T/NT) ratios at 4, 8 and 24 h post-injection were calculated and compared among different groups. One-way analysis of variance, repetitive measurement analysis of variance and Dunnett-t test were used to compare the data of different groups. The relationship between %ID/g and T/NT was analyzed with Pearson correlation. Results: The radiolabeled yield was more than 75% and radiochemistry purity was more than 90%. The difference of %ID/g at 24 h post-injection was significant (F=14.400, P<0.001) in tumors of all groups. The radioactive uptake in tumor of ¹³¹I-PAMAM(G5.0)-SR group was the highest at 24 h post-injection((1.80±0.18) %ID/g). There were significant differences of T/NT ratios among different groups(F=4.776, P<0.05)and between different time points(F=8.630, P<0.05). Compared with negative control group (Na¹³¹I), the T/NT ratios significantly increased in ¹³¹I-PAMAM(G5.0)-SR group at 4, 8 and 24 h post-injection (t=4.169, 7.123 and 4.032, all P<0.05) and in ¹³¹I-PAMAM(G5.0)-GP group at 4 h post-injection (t=5.893, P<0.05). The T/NT ratio in ¹³¹I-PAMAM(G5.0)-SR group was higher than that in ¹³¹I-PAMAM(G5.0)-GP group at 24 h post-injection (t=2.871, P<0.05). Conclusions PAMAM(G5.0)-SR, PAMAM(G5.0)-GP and PAMAM(G5.0)-SR/GP can target the MTC models. ¹³¹I-PAMAM(G5.0)-SR has the best biological properties and may provide a new precision method for MTC diagnosis, treatment and prognosis evaluation.

Objective To explore the clinical technical points of the treatment of soft tissue defect of the foot and ankle with the supercharged peroneal artery perforator propeller flap,and to provide theoretical support by anatomical observation.Methods From January,2010 to February,2018,a total of 10 patients with soft tissue defect of foot and ankle were treated with supercharged peroneal artery perforator propeller flap.Cause of injury:trauma in 7 cases,wound ulcer in 1 case,and poor healing of the calcaneus incision in 2 cases.Defect site:5 cases of heel,2 cases of medial and lateral malleolus,and 3 cases of dorsum and sole.The size of flap ranged from 6.0 cm×3.0 cm to 16.0 cm×5.0 cm.All patients were followed-up at 1,3,6 months after operation,and the function recovery was judged by AOFAS Ankle Hindfood Scale at 3 months post-opertively.From November,2016 to May,2017,the anatomical basis and operative points of the supercharged peroneal artery perforator flap were summarized.Results All the 10 cases of supercharged peroneal artery perforator propeller flap survived.Two of them had local epidermal necrosis at the proximal end of the flap.After 1 to 2 weeks of dressing,they finally healed.The other 8 cases healed well.Anatomical studies showed that different planes of the supercharged peroneal artery perforator propeller flap can only reduce the compression of the double pedicles and reduce the distal necrosis rate of the flap by rotating in different rotation directions.Conclusion The supercharged peroneal artery perforator propeller flap can enhance the blood supply and venous return in the "big paddle" artery of the flap,preventing distal necrosis.

Objective To evaluate the feasibility and clinical efficacy of our self-designed simple skin stretching device combined with collagen sponge for management of severe soft tissue wounds.Methods From September 2015 to October 2017,a consecutive series of 43 patients whose soft tissue wounds could not be closed primarily were enrolled for a therapy using a simple skin stretching device made of round osseous pins and wire combined with collagen sponge.They were 27 males and 16 females,with a mean age of 31.5 years (from 5 to 56 years).There were 18 fresh wounds and 25 old ones.Their skin defects ranged from 5.5 cm × 3.0 cm to 18.0 cm × 7.5 cm.After debridement and vacuum sealing drainage,2 round osseous pins with a diameter of 2.0 mm or 2.5 mm were driven through the dermis about 1 to 2 cm from both edges of the wound,in parallel with the longitudinal axis of the wound.After the parts of 2 pins exposed outside the skin were bent,they were fixed respectively with a fine wire with 2 twisted strands.The wounds were continuously stitched with eversion suture.The wires and sutures were gradually tightened to contract the wounds until the skin color changed and capillary filling reaction started.Then medical collagen sponge was used to cover the wounds.Next,the wires and sutures were tightened continuously until the wound edges were pulled together.Details of this therapy and its complications were recorded.Follow-up visits were paid until wound healing.Results Of the 43 cases,the wounds were directly closed immediately after primary stretching procedure in 8,closed after skin stretching for 4 to 12 days (average,7.5 days) in 30,and significantly reduced in 5 which were cured following skin graft.Eventually,40 cases were followed up for an average of 6.8 months (from 3 to 18 months) and 3 were lost.Aesthetic reoperation was performed in 3 patients who were inflicted with postoperative scar formation after skin graft.Linear healing of the wound edges was achieved in 37 patients without complications like skin necrosis,pathological hyperplasia scar,skin sensation deletion or wound infection,leading to fine appearance and functional recovery.Conclusion Our self-designed simple skin stretching device combined with collagen sponge provides a cost-effective and practical technique for clinical treatment of soft tissue defects,with an advantage of reducing or even avoiding secondary repair with skin graft or skin flap.

Aim To establish and optimize the VSVG/HIV-1NL4-3 Luc pseudovirus model for anti-HIV drugs screening. Methods The infectivity of VSVG/HIV-1 NL4-3 Luc in 4 different cell lines was investigated according to the method of the lucifer-ase activity analysis system of Promega company. 3 different ex-perimental settings were used to detect the activities of approved anti-HIV drugs to confirm the feasibility and effectiveness of the system. Finally, some potential compounds were screened for their anti-HIV activities, and their antiviral activities against the pseudovirus were compared with HIV-1ⅢB . Results The pseud-ovirus showed the strongest replication ability in CRFK cells, and a clear dose-effect relationship was found between the report gene expression level and the virus quantity. Comparing the EC50 of different positive inhibitors against VSVG/HIV-1 NL4-3 Luc on 3 kinds of experimental conditions, 3rd scheme is the best. Finally, the system was used to screen compounds, the EC50 s a-gainst pseudovirus were similar to those in HIV-1ⅢB . Conclusion An optimized VSVG/HIV-1 NL4-3 Luc anti-HIV screening sys-tem has been successfully developed.

Background and purpose:Researches had indicated that about over 85%of malignant tumors highly express telomerase activity. So telomerase has become one of the important methods in the research field of tumor diagnosis and treatment. Nowadays, several reports about malignant tumor which over expresses hTERT targeting imaging with radionuclide labeled hTERT ASON had been published. In these reports, high quality of pictures can hardly be acquired because of poor anatomical and spacial resolution in nuclear imaging itself. Accordingly, in this study, we developed a method of detecting human telomerase in vivo with magnetic resonance imaging (MRI) and evaluate its feasibility. Methods:Firstly, Uniformly phosphorothioate-modified human telomerase reverse transcriptase antisense oligonucleotide (hTERT ASON) was labeled with Gd3+ through the bifunctional chelator 1, 4, 7, 10-tetraazacyclododecane-N, N’, N’’, N’’’-tetraacetic acid (DOTA) and iv vitro experiments were performed to characterize the antisense probes (for biodistribution and cellular uptake, 99mTc-DOTA-ASON was used in stead of Gd-DOTA-ASON). Then Gd-DOTA-ASON was injected intraperitoneally in pulmonary adenocarcinoma A375 nude mice tumor-bearing BALB/c for in vivo imaging using 7.0 T Micro MRI periodically, tumors and their surrounding tissues were defined as region of interest (ROI) to calculate the signal to noise ratio (SNR) of tumor to muscle using Gd-DTPA as control. Finally, immunohistochemical analysis of telomerase activity of each xenograft was operated 2 days after imaging. Results:The binding efficiency of Gd-DOTA-ASON reached was as high as 65%(63.2±2.4, n=6). And it can maintain 61%in fresh human serum and normal saline at 37℃over 24 h;A375 cells showed an uptake of 8.5%when incubated with 99mTc-DOTA-ASON;In comparing with DOTA-ASON and Gd-DTPA, cells transected with Gd-DOTA-ASON had higher SI when performed MRI with T1WI. The hTERT-expressing xenografts were obviously enhanced by Gd-DOTA-ASON at 0.5-6 h after injection and the SNR can reach 2.37, whereas obvious enhancement only could be found within 2 h after injection of Gd-DTPA. Both labeled and non-labeled antisense probes can suppress the activity of telomerase of A375 cells either in vitro or in vitro. Conclusion:Our research offers proof that Gd-DOTA-ASON can be used as tumor specific targeting MR probe for diagnosing malignant tumors with high expression of telomerase.