BACKGROUND:Titanium implants are widely used in clinical practice because of their high strength and good biocompatibility.However,during implantation,bacterial infection and tissue damage environment produce a large number of reactive oxygen species,which can easily lead to delayed tissue healing and surgical failure.Consequently,the development of titanium implants with antimicrobial and antioxidant properties becomes paramount. OBJECTIVE:Considering the potent antimicrobial attributes of copper ions and the remarkable antioxidant qualities of polyphenols,we proposed the fabrication of polyphenol-mediated copper ion coatings on titanium surfaces.These coatings were subsequently assessed for their in vitro antimicrobial and antioxidant properties. METHODS:Nanostructures were generated on the titanium surface using the alkali thermal method.The titanium was immersed in a solution containing tannic acid and copper ions to achieve polyphenol-mediated copper ion coatings.The surface morphology and water contact angle were detected.The loading and release of copper ions were examined using atomic absorption spectroscopy.Staphylococcus aureus was inoculated on the surface of pure titanium sheet(blank group),alkali heat treated titanium sheet(control group),and polyphenol mediated copper ion modified titanium sheet(experimental group)to observe the bacterial survival status.Osteoblast precursor cells MC3T3-E1 were co-cultivated on the surface of three groups of titanium sheets to assess their antioxidant properties and bioactivity. RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the polyphenol-mediated copper ion modified titanium sheet had rod-like nanostructures and no cracks on the surface.The surface hydrophilicity of copper ion modified titanium sheet mediated by polyphenol was close to that of pure titanium sheet.Atomic absorption spectrometry results showed a 51%increase in the loading capacity of copper ions after polyphenol mediation,with a uniform release of copper ions.(2)The antibacterial rates of titanium sheets in the blank group,control group,and experimental group were 0%,21.65%,and 93.75%,respectively.The live/dead staining and CTC staining showed that the live bacteria on the surface of titanium plates in the blank group were the most,and the live bacteria on the surface of titanium plates in the experimental group were the least.(3)The results of live/dead staining and CCK-8 assay showed that the three groups of titanium sheets had good cytocompatibility,and the titanium sheets in the experimental group were more conducive to the proliferation of MC3T3-E1 cells.Active oxygen fluorescence probe detection exhibited that compared with the other two groups,the fluorescence intensity of active oxygen on the surface of the experimental group was significantly reduced.The results of alkaline phosphatase and alizarin red S staining showed that the osteogenic differentiation and extracellular matrix mineralization of MC3T3-E1 cells on the surface of titanium sheets in the experimental group were stronger than those in the other two groups.(4)These results show that the polyphenol-mediated copper ion coating has strong antibacterial and antioxidant properties and promotes osteogenic differentiation.

Gene synthesis is an enabling technology that supports the development of synthetic biology. The existing approaches for de novo gene synthesis generally have tedious operation, low efficiency, high error rates, and limited product lengths, being difficult to support the huge demand of synthetic biology. The assembly and error correction are the keys in gene synthesis. This study first designed the oligonucleotide sequences by reasonably splitting the virus genome of approximately 10 kb by balancing the parameters of sequence design software ability, PCR amplification ability, and assembly enzyme assembly ability. Then, two-step PCR was performed with high-fidelity polymerase to complete the de novo synthesis of 3.0 kb DNA fragments, and error correction reactions were performed with T7 endonuclease Ⅰ for the products from different stages of PCR. Finally, the virus genome was assembled by 3.0 kb DNA fragments from de novo synthesis and error correction and then sequenced. The experimental results showed that the proposed method successfully produced the DNA fragment of about 10 kb and reduced the probability of large fragment mutations during the assembly process, with the lowest error rate reaching 0.36 errors/kb. In summary, this study developed an efficient de novo method for synthesizing a viral genome of about 10 kb with T7 endonuclease Ⅰ-mediated error correction. This method enabled the synthesis of a 10 kb viral genome in one day and the correct plasmid of the viral genome in five days. This study optimized the de novo gene synthesis process, reduced the error rate, simplified the synthesis and assembly steps, and reduced the cost of viral genome assembly.
Genome, Viral/genetics* ; Polymerase Chain Reaction/methods* ; DNA, Viral/genetics* ; Bacteriophage T7/enzymology* ; Synthetic Biology/methods*

Objective:To explore the effect of accurately localized mini anterolateral thigh perforator flap in repairing medium-sized skin and soft tissue defects in fingers.Methods:The study was a retrospective observational study. From December 2019 to September 2022, 15 patients with medium-sized skin and soft tissue defects who met the inclusion criteria in fingers were admitted to the Second Affiliated Hospital of Wenzhou Medical University, including 12 males and 3 females, aged 23 to 62 years. After debridement, the wounds were all accompanied by exposed tendons, bones, vessels and nerves, with an area from 4.0 cm×3.0 cm to 8.0 cm×3.5 cm. Computed tomography angiography and color Doppler ultrasonography examinations were performed on both lower limbs of the patient before surgery to accurately locate the anterolateral thigh perforators. When the flap with area from 6.0 cm×3.0 cm to 11.0 cm×4.0 cm was harvested, the flap was thinned. The artery and vein perforators of the flap were anastomosed respectively with the digital artery and dorsal metacarpal vein. If there was avulsion injury, infection, or burn in the recipient area, the main arterial and veinous vessels carried by the skin flap was anastomosed with the radial artery and accompanying vein. The lateral thigh cutaneous nerve carried by the flap was anastomosed with the stump of the digital nerve. The types of perforators of the lateral thigh artery were observed during operation and compared with the location of the vessels before operation. After operation, the survival and adverse complication of the flap were closely observed. During follow-up, the skin flap color, texture, and shape were observed; the wound healing in donor area was observed. At the last follow-up, the two-point discriminative distance of the affected finger pulp was measured, and the function of the affected finger was evaluated using the trial standard for the evaluation of functions of upper limbs of Hand Surgery Society of Chinese Medical Association, and the interphalangeal joint movement of the affected finger was observed; the patients' complaints about the adverse effects of flap resection on lower limbs were recorded.Results:During the operation, it was observed that the perforators of the flaps in 11 patients were the descending branch of the lateral circumflex thigh artery, in two patients, the perforators of skin flaps were the oblique branch of the lateral thigh artery, and the perforators in another two patients were the transverse branch of the lateral circumflex thigh artery, which were consistent with the preoperative vascular localization. After operation, all flaps survived without vascular crisis and infection. The patients were followed up for 6-12 months, the flaps had excellent color, texture, and appearance; only linear scars remained on the donor wound. At the last follow-up, the two-point discrimination distance in the finger pulp was 7-11 mm; the affected finger function was rated as excellent in 6 cases, good in 6 cases, and fair in 3 cases; the flexion and extension function of the finger was not affected; two patients complained of numbness in the lateral thigh after excision of the skin flap, and the other 13 patients had no complain of adverse complaints.Conclusions:The perforating branch in lateral thigh region can be accurately located by computed tomography angiography and color Doppler ultrasonography, accurate positioning of perforators before operation can reduce the damage to the donor area during the incision of the flap, the appearance and function of the affected finger can be restored to the maximum extent by thinning the transplanted flap and rebuilding the finger sensation. Therefore, it is an effective and reliable way to repair the medium-sized skin and soft tissue defects of fingers with the mini thigh anterolateral perforator flap.

Objective:To compare the effects of two methods of marking surface landmarks on the patient’s positional stability when using a multifunctional body board in combination with thermoplastics to fix the abdominal and pelvic areas for radiotherapy patients.Methods:50 subjects who underwent positional fixation using a multifunctional body board in combination with thermoplastics from August 2022 to January 2023. The subjects were divided into two groups, A and B, with 25 cases each, according to the different methods of body surface marking. In group A, landmarks were marked on the body surface on the top edge of the thermoplastics. In group B, three sets of surface landmarks were marked on the patient’s body according to the laser line on the projection of the patient’s body surface when the thermoplastics were completed. Manual registration is performed using L3 to L5 as the main registration targets. The pre-treatment CBCT image is used to analyze the first-time positioning pass rate, setup errors in the x-, y-, and z-axis directions, and the distribution of positive and negative setup errors in both groups of patients. Results:The pass rates of the first-time positioning of patients in Groups A and B were 76.9% and 86.1%, respectively, which met the clinical requirements. Group B had a better first-time positioning pass rate than group A, and the difference between the two groups was statistically significant ( P < 0.05). The pendulum errors of group B were smaller than those of group A in both the x-axis and y-axis (all P < 0.05), and the difference between the two groups in terms of the pendulum errors in the z-axis direction was not statistically significant (all P > 0.05). The difference in the frequency distribution of the pendulum error in the positive and negative directions of the x- and z-axis between the two groups was not statistically significant (all P > 0.05). The difference in the frequency of distribution of the pendulum error in the positive and negative directions of the y-axis between the two groups was statistically significant ( P < 0.05). Conclusions:The proposed two methods of surface landmark marking are generally in line with the positioning requirements for conventional fractionation radiotherapy for abdominal and pelvic patients. Using a laser line on the projection of the patient’s body surface for three sets of surface landmark markings produces smaller setup errors and is better than using the top edge of the thermoplastics for surface landmark markings, improving the positional stability of abdominal and pelvic patients.

Objective:To investigate the effects of different radiotherapy regimens on the short-term and long-term prognosis of patients with limited-stage small cell lung cancer (SCLC).Methods:Sixty patients with limited-stage SCLC in the Third People′s Hospital of Yangquan City from September 2017 to September 2019 were selected. The patients who received concurrent radiotherapy and chemotherapy were in group A and the patients who received sequential radiotherapy and chemotherapy were in group B, 30 cases in each group. The short-term efficacy and adverse reactions of the two groups were compared, and the levels of neuron-specific enolase (NSE), gastrin release precursor (ProGRP), carcinoembryonic antigen (CEA), vascular endothelial growth factor (VEGF), endostatin (ES), circulating endothelial cells (CEC) were compared between the two groups. The survival status of the two groups were followed-up.Results:After treatment, the disease control rate in group A was higher than that in the group B : 80.00%(24/30) vs. 53.33%(16/30), the difference was statistically significant ( χ2 = 4.80, P<0.05). The levels of serum NSE, ProGRP and CEA in the group A after treatment were lower than those in the group B: (19.42 ± 3.31) pg/L vs. (24.58 ± 4.42) pg/L, (95.45 ± 10.33) ng/L vs. (115.54 ± 15.66) ng/L, (8.25 ± 1.02) μg/L vs. (10.33 ± 1.15) μg/L, the differences were statistically significant ( P<0.05). The levels of VEGF and CEC in the group A were lower than those in the group B after treatment and ES was higher than that in the group B: (356.62 ± 56.63) ng/L vs. (442.21 ± 55.38) ng/L, (65.56 ± 5.41) × 10 6/L vs. (99.28 ± 7.24) ×10 6/L, (52.65 ± 6.44) μmol/L vs. (31.85 ± 5.49) μmol/L, the differences were statistically significant ( P<0.05). There was no significant difference in the incidence of adverse reactions between the two groups ( P>0.05). The survival rate in the group A was higher than that in the group B at 1 year after treatment: 82.14%(23/28) vs. 56.00%(14/25), the difference was statistically significant ( χ2 = 4.28, P = 0.038). Conclusions:Compared with sequential radiotherapy and chemotherapy, concurrent radiotherapy and chemotherapy is effective in treating limited-stage SCLC, which can adjust the level of tumor markers and angiogenesis indicators, and improve the treatment effect and survival rate.

Resource utilization is an effective way to cope with the rapid increase of kitchen waste and excess sludge, and volatile fatty acids produced by anaerobic fermentation is an important way of recycling organic waste. However, the single substrate limits the efficient production of volatile fatty acids. In recent years, volatile fatty acids produced by anaerobic co-fermentation using different substrates has been widely studied and applied. In this paper, we analyze the characteristics of fermentation to produce acid using kitchen waste and excess sludge alone or mixture. Influences of environmental factors and microbial community structure on the type and yield of volatile fatty acids in the anaerobic fermentation system are discussed in detail. Moreover, we propose future research directions, to provide a reference for recycling kitchen waste and excess sludge.
Anaerobiosis ; Bioreactors ; Fatty Acids, Volatile ; Fermentation ; Hydrogen-Ion Concentration ; Microbiota ; Organic Chemicals ; Sewage

OBJECTIVE: To analyze the clinical manifestations of heart, liver and kidney damages in the early stage of COVID-19 to identify the indicators for these damages. METHODS: We analyzed the clinical features, underlying diseases, and indicators of infection in 12 patients with COVID-19 on the second day after their admission to our hospital between January 20 and February 20, 2020.The data including CK-MB, aTnI, BNP, heart rate, changes in ECG, LVEF (%), left ventricular general longitudinal strain (GLS, measured by color Doppler ultrasound) were collected.The changes of liver function biochemical indicators were dynamically reviewed.BUN, UCR, eGFR, Ccr, and UACR and the levels of MA, A1M, IGU, and TRU were recorded. RESULTS: The 12 patients included 2 severe cases, 8 common type cases, and 2 mild cases.Four of the patients presented with sinus tachycardia, ECG changes and abnormal GLS in spite of normal aTNI and LVEF; 1 patient had abnormal CKMB and BNP.On the first and third days following admission, the patients had normal ALT, AST and GGT levels.On day 7, hepatic function damage occurred in the severe cases, manifested by elevated ALT and AST levels.Abnormalities of eGFR, Ccr and UACR occurred in 8, 5 and 5 of the patients, respectively.Abnormal elevations of MA, A1M, IGU and TRU in urine protein were observed in 4, 4, 5, and 2 of the patients, respectively. CONCLUSIONS In patients with COVID-19, heart damage can be identified early by observing the GLS and new abnormalities on ECG in spite of normal aTNI and LVEF.Early liver injury is not obvious in these patients, but dynamic monitoring of the indicators of should be emplemented, especially in severe cases. In cases with normal CR and BUN, kidney damage can be detected early by calculating eGFR, Ccr and UACR and urine protein tests.
Betacoronavirus ; COVID-19 ; Coronavirus Infections ; Humans ; Pandemics ; Pneumonia, Viral ; SARS-CoV-2

Objective To investigate the effects of enteral ecological nutrition on intestinal immune function and Hh protein expression in intestinal mucosa of rats with small intestinal injury and explore the mechanism. Methods Thirty male clean grade Wistar rats were selected as study subjects. The rats were randomly divided into a model group and an enteral ecotrophic group with 15 rats in each group. The small intestinal injury model was prepared by trauma method. After successful modeling, Six hours after successful establishment of the model, the rats in the two groups were fed with 753.12 kJ·kg-1·d-1 energy, 3 times a day. The rats in the model group were fed with conventional diet, while the rats in the enteral ecotrophic group were fed with enteral nutrition emulsion +Lactobacillus, bifidobacterium triple viable bacteria by intragastric administration of 1×107 cfu/d. After 10 days, the rats were killed, the small intestine of the two groups was dissected and stained with hematoxylin-eosin (HE) staining, and the morphological changes of small intestinal mucosa (villus height, glandular recess depth, mucosal thickness) were observed in the two groups; the expressions of CD3+, CD4+, CD8+ positive T cells in small intestinal mucosa were evaluated by immunohistochemistry; the expression of Hh protein in small intestinal mucosa was detected by Western Blot in the two groups. Results On the first day after the establishment of the model, the weight of rats in both groups was lower than that before the modeling [model group (g): 118.0±4.2 vs. 121.7±5.2, enteral ecotrophic group (g):117.5±4.7 vs. 120.8±5.0, P > 0.05], from the fifth day after the modeling, the weight of the rats in the enteral ecotrophic group was significantly higher than that of the model group (g: 127.1±5.0 vs. 123.2±4.2, P < 0.05), continued to 10 days (g: 142.5±6.6 vs. 135.3±6.2, P < 0.05). After the establishment of the model for 10 days, the small intestinal villus height, glandular recess depth, mucosal thickness and percentages of CD3+, CD4+ and CD8+ positive T cells in enteral ecotrophic group were significantly higher than those in model group [villus height (μm): 221.7±25.0 vs. 159.5±20.8, glandular recess depth (μm): 79.39±12.65 vs. 67.87±7.79, mucosal thickness (μm): 254.7±51.8 vs. 209.0±27.2, CD3+: 0.193±0.035 vs. 0.125±0.031, CD4+: 0.130±0.027 vs. 0.104±0.015, CD8+: 0.165±0.026 vs. 0.137±0.027, all P < 0.05]. The expression of Hh protein in the enteral ecotrophic group was obviously higher than that in model group (Hh/β-actin: 0.16±0.04 vs. 0.04±0.02, P <0.05). Conclusion Enteral ecological nutrition may promote the repair of intestinal mucosa and the improvement of immune function level by enhancing the expression of Hh protein in small intestinal mucosa of rats with small intestinal injury.

Objective To discuss the safety and the application of the self-designed multifunctional inflatable pelvis and hip-joint fixator (MIPHF) in damage control in pelvic fracture patients.Methods The MIPHF was subjected to pressure test and quality inspection.From September 2016 to June 2017,61 pelvic-fracture patients were treated with our self-designed MIPHF as pre-hospital first-aid care according to the concept of damage control orthopedics (DCO) (MIPHF group).The control group consisted of 69 pelvic-fracture patients who had not received pre-hospital first-aid care with the self-designed MIPHF from December 2015 to August 2016.There were no statistically significant differences between the two groups in gender,age,types of pelvic fracture,and preoperative injure severity score (ISS).The study compared the two groups for the case fatality rate,volume of blood transfused during surgeries,early complication rates,fracture reduction (Matta standards),and long-term efficacy (Cole scores).Results The pressure test showed that the MIPHF had a good fixation effect on the pelvis.And the quality inspection showed that the material used for the MIPHF was in line with national standards and the safety was guaranteed.The MIPHF group had 1 death (1.6％) and the control group had 8 deaths (11.6％),which was a significant difference (x2=4.979,P=0.026).All survival patients in both groups were followed up.The MIPHF group (61 cases) received 3.0 to 18.0 months follow-up,with an average of 9.0 months.And the control group (69 cases) had 18.0 to 30.0 months follow-up,with an average of 21.9 months.In the MIPHF group,23 cases were treated conservatively,and 37 cases were treated with surgery.Among them,3 cases were fixed with external fixator,20 cases with anterior open reduction and internal fixation,9 cases with posterior open reduction and internal fixation,and 5 cases with combined anterior and posterior fixation.The timing of surgery was 1 to 20 days after injury,with an average of 4.1 days.The volume of blood transfused in the MIPHF group during surgery was 200 to 1500 ml,with an average of 628.6 ml.In the control group,27 patients were treated conservatively,and 42 patients were treated with surgery.Among them,2 cases were fixed with external fixator,24 cases with anterior open reduction and internal fixation,10 cases with posterior open reduction and internal fixation,and 6 cases with combined anterior and posterior fixation.The timing of surgery was 1 to 15 days after injury,with an average of 3.l days.The volume of blood transfused in the control group during surgery was 200 to 4000 ml,with an average of 1 707.1 ml.There was a significant difference between the two groups in intraoperative blood transfusion(Z=-2.330,P=0.020).The MIPHF group had 10 (16.4％) cases of early serious complications and the control group had 22 (31.9％) cases,which had a significant difference (x2=4.187,P=0.041).According to the criteria proposed by Matta et al.,the good rate of results for treating fractures was 82.0％ in the MIPHF group and 60.9％ in the control group,which got a significant difference (x2=6.967,P=0.008).The MIPHF group and the control group also differed significantly in their mean long-term Cole scores (27.2±4.0 versus 25.1 ±5.6,t=2.457,P=0.015).Conclusion MIPHF,which reflects the DCO concept,may be recommended as pre-hospital first-aid care for patients with pelvic fracture because it can lessen bleeding and prevent secondary pelvic injury,thus reduce case fatality rate and the incidence of complications.It can also improve the success rate of treating pelvic fracture,which will positively affect long-term outcomes.

Background and purpose: The prognosis of completely resected stage ⅢA(N2) non-small cell lung cancer (NSCLC) remains a significant concern. The 5-year overall survival (OS) rates range from 10% to 30%. This study aimed to analyze the patterns of first failure in completely resected stage ⅢA(N2) NSCLC and to assess the actuarial risk of developing metastasis at different sites and to guild standard clinical practice. Methods: Patients withⅢA(N2) NSCLC who had undergone radical surgery in our hospital from Jan. 2005 to Jul. 2012 were retrospectively reviewed. The progression-free survival (PFS), the OS, patterns of first failure, the actuarial risk were analyzed. The cumulative incidence of first failure was determined using the Kaplan-Meier analysis. Results: Among 357 patients who met the eligibility criteria with completely resected stage ⅢA(N2) NSCLC, 5-year OS was 36.9%. There were 284 (77.6%) patients experiencing disease failure: 61 with local failure, 197 with local and distant failures, and 26 patients with local recurrence as the first failure. Brain, bone and lung were the main sites of distant failure as the first failure, while brain was the most common site. There were 67 patients developing brain metastases (BM) as the first site of failure. The median time of local failure as the first site of failure was 13.6 months, and the time to develop distant recurrence was 15.1 months. 92.5% BM developed in 3 years after the complete resection. Conclusion: As the first failure, the rate of distant failure was much higher than that of local failure in completely resected stage ⅢA(N2) NSCLC. Brain was the most common site of distant failure as the first failure. These results can be helpful in guiding standard clinical practice and evaluating the outcome of comprehensive treatment.