Objective To investigate the effects of human Rrain-derived neurotrophic factor-gamma fetopro-tein (hBDNF-GFP) gene-transfected neural stem cell (NSC) transplantation on BDNF expressions in the retina of rots after optic nerve (ON) crush injury. Method ①Seventy-eight Sprague-Dawley (SD) rats were randomly as-signed into a control group (n = 6) and ON crush group (n = 72). In the ON crush group, the right ON was crushed while the left NO was exposed as sham injury. Rats in the ON crush group were divided into three sub-groups: PBS group (intravitreons injection of 0.01 mol/L phosphate buffered solution); GFP group (intravitreous transplantation of GFP gene-transfected NSCs); and hBDNF-GFP group (intravitreous transplantation of hBDNF-GFP gene-transfected NSCs). Rats were sacrificed 3, 7, 14 and 28 days after transplantation, and BDNF expres-sions in retinal homogenates was detected by using enzyme-linked immunosorbent assay (ELISA). ②The hBDNF-GFP-NSCs were transplanted intravitreous into six rats after ON crush injury. Following this, two rats were sacri-riced 2, 4 and 8 weeks after transplantation. The survival and location of NSCs in host retina were observed by frozen section analysis. ③Adult SD rats were randomly divided into four groups: control group (n = 5); NSC group (NSC transplantation, n = 10); GFP-NSC group (GFP-NSC transplantation, n = 10); and hBDNF-GFP-NSC group (hBDNF-GFP-NSC transplantation, n = 10). Four and eight weeks after transplantation, five rats from every group were sacrificed. Western blot analysis was used to determine retinal BDNF expression. Results ① There was no significant difference in BDNF expression between the control group and sham-injury groups (P >0.05). Three days after NSC transplantation, BDNF expression increased significantly in the three injured sub-groups compared with the sham-injury group, (P < 0.05), whereas no significant inter-group differences in BDNF expressions among three injured sub-groups were observed (P > 0.05). Seven days after transplantation, there was a significant difference in BDNF expression between the GFP-NSC group and the sham-injury groups (P <0.05), whereas there were no significant differences in BDNF expressions among the PBS, hBDNF-GFP-NSC and sham-injury groups (P > 0.05). Fourteen and 28 days after transplantation, BDNF expressions decreased in the PBS group and the GFP-NSC groups, while BDNF expressions in the hBDNF-GFP-NSC group increased significant-ly compared with the other three groups (P < 0.05); ②Frozen section showed that transplanted hBDNF-GFP-NSCs could survive and gradually extended to all layers of the host retina. ③Westem blot revealed there were no differences in BDNF expressions between 4-week and 8-week intervals in the hBDNF-GFP-NSC group. Compared with other three groups, BDNF expressions in the retina increased significantly after hBDNF-GFP-NSC transplanta-tion. Conclusions The hBDNF-GFP gene-trausfected NSCs can survive in the host retina and BDNF expressions are stable at a high level.

Objective To investigate the changes of expression of stromal cell-derived factor 1α (SDF-1α) in retinas after partial optic nerve injury in rats. Methods Models with injury of partial optic nerve were induced in rats. Retinal tissues were collected 1,2,3,5,7,10 and 14 d after injury. Enzyme linked immunosorbent assay and Real-time quantitative PCR were employed to detect the expression of SDF-1α protein and mRNA in retinal tissues respectively in injury group (n=28), sham operated group (n=28) and normal control group (n=12). Results The expression of SDF-1α protein and mRNA in retinas was higher than that in sham operated group and normal control group at different time points after injury (P<0.01), and it reached the peak at the 5th day after injury. The expression of SDF-1α protein and mRNA maintained a high level at the 14th day after injury. Conclusion The expression of SDF-1α protein and mRNA is up-regulated after partial optic nerve injury, and may last for a long time.

Objective To explore diagnostic value of amplitude of low-frequency fluctuation ( ALFF) on cognitive impairment in amyotrophic lateral sclerosis ( ALS ) using resting-state functional magnetic resonance imaging ( MRI ) .Methods Sixteen ALS patients from neurological clinic in Peking Union Medical College Hospital were enrolled between November 2013 and April 2015.The patients were divided into two groups by the presence (ALSi, n=7) or absence (ALSu, n=9) of cognitive impairment. Routine MRI structural images and resting-state functional MRI were collected for comparison between groups through voxel-based morphometry ( VBM ) and ALFF.Results ( 1 ) Neuropsychological analysis showed significant differences in Montreal Cognitive Assessment score (22.9 ±2.0 vs 25.8 ±2.3, t=2.622, P=0.020), Frontal Assessment Battery score (12.4 ±1.6 vs 15.1 ±1.4, t=3.600, P=0.003), animal listing test (13.6 ±1.8 vs 16.7 ±2.9, t=2.482, P=0.026), naming test (2(1) vs 0(1), Z=-2.746, P=0.006), similarity test (7.9 ±3.7 vs 17.3 ±2.8, t=5.846, P=0.000) and clock drawing test (2(2) vs 3(0), Z=2.516, P=0.012).(2) VBM analysis showed no significant differences in both gray matter and white matter density between the two groups .(3) ALFF analysis showed significantly increased signals in widespread areas of bilateral cerebrum and cerebellum in ALSi group compared to ALSu group . Conclusion ALFF value has the potential to provide more valuable imaging basis for early diagnosis on cognitive impairment in ALS.

Objective To evaluate clinic efficacy and safety of tigecycline combined with cefoperazone/sulbactam for treatment of intracranial infection with drug-resistant Acinetobacter baumannii (DRAB).Methods 12 patients with DRAB intracranial infection admitted in a department of neurosurgery between January 2014 and April 2015 were analyzed retrospectively,clinic efficacy and safety of tigecycline combined with cefoperazone/sulbactam for treatment of DRAB intracranial infection were evaluated.Results After patients received tigecycline combined with cefoperazone/sulbactam for 12-62 days (39.5 days on average), clinical symptoms and signs (including body temperature,signs of meningeal irritation)of most patients were significantly improved compared with before treat-ment,3 cases were cured,5 were markedly effective,4 abandoned or ineffective(death).The overall effective rate was 66.67%(8/12),mortality was 33.33% (4/12);bacterial clearance rate of cerebrospinal fluid (CSF)was 83.33%(10/12).Causes of death:2 were with failure of brain stem following craniocerebral trauma,1was extensive parenchymal infection,1was recurrence of intracranial infection and formation of brain abscess after withdrawing an-timicrobial agents.No significant adverse reaction occurred during the treatment period.Conclusion On the basis of keeping CSF drainage unobstructed,long course treatment of tigecycline combined with cefoperazone/sulbactam can effectively eliminate DRAB from CSF,and has a good safety.

Objective:To explore the clinical value of modified infrared thermal imaging assisted design of peroneal artery perforator propeller flap.Methods:From March 2019 to May 2021, tourniquet-reperfusion augmented thermal imaging method (TRATIM) was used to locate the perforating vessels in 14 patients for surgery or peroneal artery perforator propeller flap. The sensitivity, positive predictive value and detection time were calculated and compared with the perforating vessels located by color Doppler ultrasound(CDU), P<0.05 is statistially significant. Results:After operation, 13 flaps survived completely, but 1 flap with distal surface necrosis and healed after dressing change. Only one linear scar was left in 12 cases, and 2 cases healed well by skin grafts. Followed-up for 6-13 months, and showed that the colour and texture of the flaps were similar to the surrounding skin, without obvious swelling. The sensitivity of TRATIM and CDU in location of peral perforator vessels were 88.0% and 92.0%, respectively, and the positive predictive value was 93.6% and 95.8%, with no statistical difference between TRATIM and CDU( P>0.05). The time of positioning perforator was (3.71±0.80) min for TRATIM and(16.21±4.97) min for CDU, There was significant difference between TRATIM and CDU( P<0.01). Conclusion:With TRATIM, a surgeon can locate the peroneal perforator vessels simply, quickly and accurately, and help in design more accurate peroneal artery perforator propeller flap.

Objective:To investigate the effect of tourniquet-reperfusion augmented infrared thermography(TRAIRT) on locating the dominant perforator to assist design of free medial sural artery perforator flap.Methods:The data of patients with skin soft tissue defect of upper limb repaired by free medial sural artery perforator flap in Department of Hand Surgery, Ningbo Sixth Hospital from May 2019 to January 2022 were retrospectively analyzed. Color Doppler ultrasonography (CDU) and TRAIRT were used to locate the dominant perforator auxiliary flap design before surgery and after anesthesia. In the TRAIRT video, the hot spot with "early emergence, high brightness and fast expansion" was selected as the advantageous perforator. During the operation, the flap was elevated and transferred to the affected area to cover the defect wound according to the designed mark points, and fixed by the absorbable sutures with an interrupted suturing method, then end-to-end vascular anastomosis was performed successively. The donor area was sutured directly or sutured with full thickness skin grafts depending on the area of the wound. The donor and recipient areas of the flap were observed and the patients’ satisfaction was recorded. The gold standard was the actual location of the perforating vessel found during the operation. Compared with the gold standard, the location of the perforating vessel explored by TRAIRT and CDU was considered to be accurate (positive) if the distance was less than 10 mm. The sensitivity (accurate number of perforators/actual number of perforators during operation ×100%) and positive predictive value (accurate number of perforators/total number of perforators×100%) of the two methods were calculated, expressed as %, and the sensitivity of the two methods was compared by paired χ2 test. Kappa coefficient was used to analyze the consistency of the two methods to detect perforating vessels. The perforator detection time of TRAIRT and CDU were recorded, expressed as Mean±SD, and statistically analyzed by paired sample t-test. Results:A total of 23 patients were included, consisting of 14 males and 9 females, agd 21-70 years old, average age of 43 years old. The wounds were on forearm in 5 cases, wrist in 2 cases, and hand in 16 cases. The wound area was 4.5 cm × 5.5 cm-6.5 cm × 12.0 cm. Intraoperative flap incision area was 5.0 cm × 6.0cm-7.0 cm × 13.0 cm. After operation, 22 cases of flaps survived, 1 case had superficial necrosis at the distal end, which healed after repeated dressing change. The donor area of flaps healed well, with primary suture in 20 cases and full thickness skin grafting in 3 cases. Postoperative follow-up was 5-16 months (mean 8.4 months). The flaps had no bulge, swelling and abrasion. The texture and color were similar to the medial calf skin, and the scars in the donor and recipient areas were not obvious. The patients were satisfied with the recovery. Among the 23 patients, 49 perforators were found by TRAIRT, 50 perforators were found by CDU, and 53 perforators were found intraoperatively. The sensitivity of TRAIRT and CDU was 88.7% (47/53) and 90.6% (48/53), with no significant differences ( P>0.05), and the positive prediction value was 95.9% (47/49) and 96.0% (48/50) respectively. The Kappa coefficient was 0.89, indicating a good consistency between the two methods. The time required for TRAIRT to detect perforators was significantly shorter than that of CDU, with statistical significance [(6.52±2.02) min vs. (17.87±2.49) min, P<0.01]. Conclusion:TRAIRT has a good consistency with CDU in detection of medial sural artery perforator. The application of TRAIRT in designing medial sural artery perforator flap for wound repair is good, and it has advantages of short time taking, simple operation, economy and non-invasiveness.

Objective:To investigate the effect of tourniquet-reperfusion augmented infrared thermography(TRAIRT) on locating the dominant perforator to assist design of free medial sural artery perforator flap.Methods:The data of patients with skin soft tissue defect of upper limb repaired by free medial sural artery perforator flap in Department of Hand Surgery, Ningbo Sixth Hospital from May 2019 to January 2022 were retrospectively analyzed. Color Doppler ultrasonography (CDU) and TRAIRT were used to locate the dominant perforator auxiliary flap design before surgery and after anesthesia. In the TRAIRT video, the hot spot with "early emergence, high brightness and fast expansion" was selected as the advantageous perforator. During the operation, the flap was elevated and transferred to the affected area to cover the defect wound according to the designed mark points, and fixed by the absorbable sutures with an interrupted suturing method, then end-to-end vascular anastomosis was performed successively. The donor area was sutured directly or sutured with full thickness skin grafts depending on the area of the wound. The donor and recipient areas of the flap were observed and the patients’ satisfaction was recorded. The gold standard was the actual location of the perforating vessel found during the operation. Compared with the gold standard, the location of the perforating vessel explored by TRAIRT and CDU was considered to be accurate (positive) if the distance was less than 10 mm. The sensitivity (accurate number of perforators/actual number of perforators during operation ×100%) and positive predictive value (accurate number of perforators/total number of perforators×100%) of the two methods were calculated, expressed as %, and the sensitivity of the two methods was compared by paired χ2 test. Kappa coefficient was used to analyze the consistency of the two methods to detect perforating vessels. The perforator detection time of TRAIRT and CDU were recorded, expressed as Mean±SD, and statistically analyzed by paired sample t-test. Results:A total of 23 patients were included, consisting of 14 males and 9 females, agd 21-70 years old, average age of 43 years old. The wounds were on forearm in 5 cases, wrist in 2 cases, and hand in 16 cases. The wound area was 4.5 cm × 5.5 cm-6.5 cm × 12.0 cm. Intraoperative flap incision area was 5.0 cm × 6.0cm-7.0 cm × 13.0 cm. After operation, 22 cases of flaps survived, 1 case had superficial necrosis at the distal end, which healed after repeated dressing change. The donor area of flaps healed well, with primary suture in 20 cases and full thickness skin grafting in 3 cases. Postoperative follow-up was 5-16 months (mean 8.4 months). The flaps had no bulge, swelling and abrasion. The texture and color were similar to the medial calf skin, and the scars in the donor and recipient areas were not obvious. The patients were satisfied with the recovery. Among the 23 patients, 49 perforators were found by TRAIRT, 50 perforators were found by CDU, and 53 perforators were found intraoperatively. The sensitivity of TRAIRT and CDU was 88.7% (47/53) and 90.6% (48/53), with no significant differences ( P>0.05), and the positive prediction value was 95.9% (47/49) and 96.0% (48/50) respectively. The Kappa coefficient was 0.89, indicating a good consistency between the two methods. The time required for TRAIRT to detect perforators was significantly shorter than that of CDU, with statistical significance [(6.52±2.02) min vs. (17.87±2.49) min, P<0.01]. Conclusion:TRAIRT has a good consistency with CDU in detection of medial sural artery perforator. The application of TRAIRT in designing medial sural artery perforator flap for wound repair is good, and it has advantages of short time taking, simple operation, economy and non-invasiveness.

Objective:To investigate the effects of thinned anterolateral thigh perforator flaps combined with finger splitting and webplasty in sequential treatment of degloving destructive wound of total hand.Methods:This study was a retrospective observational study. From January 2012 to January 2023, a total of 15 cases who met the inclusion criteria with degloving destructive wound of total hand were admitted to Ningbo No.6 Hospital, including 10 males and 5 females, aged 17-75 years. The wounds were all combined with exposed bones or tendon. Emergency debridement and vacuum sealing drainage were performed in all cases before flap transplantation in stage Ⅰ. After thorough debridement, the wound area was 11.0 cm×3.0 cm-23.0 cm×13.5 cm. One or both anterolateral thigh perforator flaps with size of 12.5 cm×5.0 cm-25.0 cm×15.5 cm were designed, cut, and thinned to repair the skin and soft tissue defects of the hand. The donor site was sutured directly or repaired with medium-thickness skin graft from the opposite thigh. As needed, the flap was reconstructed by finger splitting and webplasty once or more times every 3 months after stage Ⅰoperation. The survival and complications of flap and wound healing at the donor site were observed after stage Ⅰoperation. The appearance of flap, two-point discrimination distance, and hand function were observed during the follow-up. At the final follow-up, the function of the affected hand was evaluated by the trial standards for evaluation of partial function of upper extremity by the Hand Surgery Society of Chinese Medical Association.Results:After the operation of stage Ⅰ, all the flaps of 15 cases of patients survived completely, including 1 case that had arterial crisis of flap but survived completely after exploration and re-anastomosis of blood vessels; all the wounds at the donor site healed. During the follow-up period of 6 to 18 months after stage Ⅰ, the flap was slightly swollen, with a little pigmentation, and the two-point discrimination distance in the finger flap was 8-11 mm. The fingers could complete the basic life actions such as flexion, extension, pinch, and grip. At the final follow-up, 3 cases were excellent, 9 cases were good, and 3 cases were acceptable in function evaluation of the affected hand.Conclusions:For degloving destructive wound of total hand, free transplantation of one or both thinned anterolateral thigh perforator flaps is used for repair in stage Ⅰ, and finger splitting and webplasty are used to reconstruct the flaps in the later stage, which can basically restore the pinch and grip function of the affected hand that is required for daily life, and is worthy of clinical promotion.

Objective:To explore the surgical techniques and effects of transfer of the free chimeric functional thoracodorsal artery perforator flap (TDAPF) with latissimus dorsi in reconstruction of dynamic muscle and soft tissue defects in forearm.Methods:From January 2014 to December 2020, a total of 13 transfer surgery of free chimeric functional TDAPF with vascularised latissimus dorsi were performed in the Department of Hand Surgery, Plastic & Reconstructive Surgery, Ningbo Sixth Hospital, to reconstruct forearm composite defects. The patients were 12 males and 1 female with an average age of 33.2 years old. They all had open forearm injuries, with 5 in the left and 8 in the right. Removal of inactivated muscles, exploration and repair of blood vessels and nerves were performed in emergency surgery, and VSD were applied after the surgery. Phase II reconstructive surgery were completed within 4 to 12 days, with 7.5 days in average. The wounds and flaps sized were 9.0 cm×8.0 cm - 21.0 cm×11.0 cm and were 10.0 cm×9.0 cm - 22.0 cm×12.0 cm, respectively. The volume of transferred muscles ranged were 9.0 cm × 2.0 cm × 1.5 cm - 19.0 cm × 9.0 cm × 1.5 cm. Free chimeric functional muscular flaps were transferred to reconstruct the musculus flexor digitorum profundus in 4 patients, the musculus extensor digitorum communis in 8 patients, the musculus flexor carpi radialis in 3 patients, and the musculus flexor pollicis longus in 1 patient. Reconstruction of both of musculus flexor carpi radialis and musculus extensor digitorum communis with 2 functional sub-blocks of latissimus dorsi were performed in 3 patients. All donor sites were closed primarily. All patients were included in the postoperative follow-up to evaluate the appearance of flaps, range of motion of the digits, recovery of muscle strength and gripping power, at the outpatient clinics or through the telephone interview.Results:A total of 12 flaps survived uneventfully after reconstructive surgery. One flap developed a vascular crisis and it was rectified after surgical exploration. Postoperative follow-up ranged from 17 to 52 months, with a mean of 34.1 months. Appearances of limbs and flaps were good without obvious bulky, hyperpigmentation or scar contracture. Four patients with reconstructed musculus flexor digitorum profundus showed muscle strength recovery of M 4, with the fingertips measured lower than 2.0 cm from the centre of palm when clenching a fist, and the average gripping strength of the hand reached 27.5% (20%-35%) to the healthy side. Five patients with reconstructed musculus extensor digitorum communis showed muscle strength recovery of M 4, and there was no obvious limitation in fingers flexion and extension, with the average gripping strength of the hand reached 75.4% (65%-80%) to the healthy side. Of the 3 patients with reconstruction of both power muscles, the recovery of muscle strength of musculus flexor carpi radialis was at M 4 in all the 3 patients, and the musculus extensor digitorum communis was at M 4 in 1 and M 3 in 2 patients. However, the patient who received reconstruction of musculus flexor pollicis had no significant recovery in muscle strength. Conclusion:Transfer of free chimeric functional TDAPF combines the benefits of a perforator flap and a functional muscle transfer together. This surgical technique can effectively reconstruct damaged muscle groups in forearm and resulting in good hand movement. Additionally, it can also restore the aesthetic appearance of forearm, hence makes it an excellent option for complex wound coverage.

Objective To characterize the brain microstructure changes of amyotrophic lateral sclerosis (ALS) patients with various levels of cognitive impairment as measured by diffusion tensor imaging (DTI).Methods A total of 55 ALS patients and 20 healthy controls (HC) were enrolled in the Department of Neurology of Peking Union Medical College Hospital From September 2013 to March 2017,and all participants underwent neuropsychological assessments and DTI scans.According to their cognitive performance,ALS patients were further subclassified into ALS with normal cognition (ALS-Cn,n =27),ALS with cognitive impairment (ALS-Ci,n =17) and ALS-frontotemporal dementia (ALS-FTD,n =11)subgroups.Comparisons of voxel-based and atlas-based fractional anisotropy (FA) and mean diffusivity (MD) data were conducted among the four subgroups.Results In the voxel-based analyses,the FA showed significant differences in cingulate gyms,corpus callosum,brain stem and cerebellum,and MD showed significant differences in bilateral frontal lobe,temporal lobe,cingulate gyms,corpus callosum,and cerebellum among the four subgroups.Besides,when compared to ALS-Ci,ALS-Cn and HC groups in the order,the areas of involvement were larger and differences were more significant in ALS-FTD group.In the atlas-based analyses,the FA and MD of the corticospinal tracts revealed no difference within the patients groups,but decreased FA and increased MD were found compared to HC group.The ALS-IFD patients manifested widespread white matter fiber integrity damage and microstructure impairment in the extramotor areas compared to other three groups.Conclusion The brain white matter structural patterns of ALS patients correlate with their cognitive function,and there is a gradient of alterations across the ALS-Cn,ALS-Ci and ALS-FTD continuum.