Objective To evaluate the similarities and differences between four-dimensional radiotherapy (4D-CT ) and active breathing control techniques(ABC) in respiratory management in lung radiotherapy ,and investigate the indications and feasibility of different breathing control techniques for different patients .Methods Twenty-one patients treated with lung radiotherapy received respiratory management .4D-CT technology was used in 11 patients ,while ABC technology was adopted in the rest 10 .The ratios of planning target volume(PGTV) to gross tumor volume(GTV)[(PGTV/GTV)] were calculated .The differences between these two respiratory management technologies were compared in terms of the PGTV ,positioning time ,planning time and treatment time to investigate the indications .Results 4D-CT technology had higher PGTV/GTV ratio ,and shorter positioning time and irradiation time than ABC technology(P<0 .05) ,but there was no significant difference in the planning time(P>0 .05) .In patients with ABC and 4D-CT technology ,objective response rates were 50 .0% ,45 .5% ,respectively ,and the radiation pneumonitis rates were 30 .0% ,27 .3% ,respectively .There was no significant difference in both groups (P>0 .05) .Conclusion In lung tumor radiothera-py ,ABC can reduce irradiation volume ,suitable for patients with good performance status .4D-CT is time-saving and well tolerated , suitable for patients with smaller tumors .

Objective To develop a multiplex polymerase chain reaction (PCR )method for detecting and genotyping moxifloxacin-resistant Clostridium difficile (C.difficile)isolates.Methods Specific PCR primers of slpA genotypes gr,hr,fr,gc08 and 078 were designed according to the differences of slpA nucleotide sequences in different C.difficile genotypes,and the house-keeping gene tpi specific PCR primers were also added for the construction of multiplex PCR method.Nine common intestinal normal and pathogenic strains were used to verify the specificity of slpA multiplex PCR for the detection of C.difficile.Forty-six C.difficile reference strains,belonging to 11 slpA genotypes,were used to verify the ability of the multiplex PCR method for dectecting and genotyping.Thirty-nine moxifloxacin-resistant clinical isolates were genotyped by the multiplex PCR,and its clinical value was evaluated by comparing with slpA sequence typing (slpA ST)method.Results All the 9 intestinal normal and pathogenic strains were negative when detected by the multiplex PCR.And tpi of 46 C. difficile reference strains were positive,and 36 strains belonging to slpA genotypes gr,hr,fr,gc08 and 078 were genotyped correctly.Other 10 strains which belonged to other 6 genotypes were non-typeable. Among 39 moxifloxacin-resistant clinical isolates,all were positive of tpi,and 32 isolates were typed correctly by the multiplex PCR method,including 22 slpA genotypes gc08,6 genotypes hr,2 genotypes fr,and 2 genotypes 078,which were consistent with slpA ST.However,7 isolates could not be typed by multiplex PCR,which were identified as other genotypes not included in the multiplex PCR by slpA ST. Conclusions A convenient and rapid multiplex PCR method for the detection of C.difficile is established successfully,which can distinguish among five slpA genotypes.slpA genotype gc08 is the common genotype of moxifloxacin-resistant clinical isolates.

Objective To investigate the genotype and variance of toxin associated genes of moxifloxacin‐resistant Clostridium difficile clinical isolates in Sydney .Methods Twenty‐two moxifloxacin‐resistant Clostridium difficile clinical isolates were collected from Sydney ,which were genotyped by using sequencer capillary gel electrophoresis based PCR‐ribotyping ,and toxin A and B cod‐ing gene tcdA and tcdB ,and binary toxin coding gene cdtA and cdtB were detected by using PCR method .Toxin regulator gene tc‐dC was analyzed by using PCR‐sequencing ,and was aligned with reference sequence of VPI 10463 (Genbank accession number :X92982) ,and the tcdC sequence types of all 22 isolates were identified by using blast tool in NCBI .Results Twenty‐one isolates were genotyped as hypervirulent PCR‐ribotypes 027 (RT027) ,and one isolate as RT078 ;all 22 isolates contained tcdA and tcdB for toxin A and B and cdtA and cdtB for binary toxin (tcdA+ tcdB+ cdtA+ cdtB+ ) .The tcdC sequence types of the 21 RT027 i‐solates belong to sc1 ,and that of the one RT078 isolate belongs to WA39 .Compared with tcdC reference sequence of VPI 10463 ,a consecutive 18 bp deletion (nt341 to 379) and one nucleotide deletion at position 117 were found in the 21 RT027 isolates ,and a consecutive 39 bp deletion (nt330 to 368) and one nucleotide mutation at position 184(C> T) were found in the one RT078 isolate . Conclusion Clostridium difficile hypervirulent RT027 was the common moxifloxacin resistant genotype ;Clostridium difficile hy‐pervirulent RT027 and RT078 clinical isolates contained genes for toxin A and B and binary toxin ,and contained gene sequence mu‐tation in toxin regulator gene tcdC .

Objective To develop a digital polymerase chain reaction (PCR) ribotyping method and database for Clostridium difficile genotyping.Methods Sequencer based fluorescence capillary gel electrophoresis was used,instead of agarose gel electrophoresis,to establish the digital PCR-ribotyping of Clostridium difficile.Forty Clostridium difficile reference strains,consisting of 10 PCR-ribotypes (RT),were genotyped by the new digital PCR-ribotyping method to set-up the database.Results The sequencer based fluorescence capillary gel electrophoresis correctly detected PCR-ribotyping products of the 40 reference strains,and showed as digital figure;significant differences of these digital figures were found between the 10 RT.High similar digital figures were shown in twenty-one RT027 strains,three RT002 strains and two RT014 strains.However,seven RT001 strains were typed as four subtypes,and two RT014 strains as two subtypes,respectively.Conclusion A digital PCR-ribotyping,and a reference database consisting of 10 RT are successfully established.

Objective To investigate the genotype and production of toxin A and B of C .difficile clinical isolates collected from Sydney ,Australia .Methods Sixty‐eight C .difficile clinical isolates were collected from Westmead Hospital ,the University of Sydney ,which were genotyped by using PCR‐ribotyping ,and toxin A ,B coding gene tcdA ,tcdB were detected by using PCR meth‐od .Results Thirty‐one PCR‐ribotypes (RTs) were confirmed in the 68 C .difficile clinical isolates ,RT014 (19 .1% ) and RT002 (11 .8% ) were the common genotypes .Sixty‐four of 68 (94 .1% ) isolates contained tcdA and tcdB for toxin A and B .Conclusion The common prevalent PCR‐ribotypes of C .difficile were RT014 and RT002 in Sydney ,most of the C .difficile clinical isolates contained toxin A and B .

Objective To investigate the carrying status and characteristics of Clostridium difficile isolated from infants.Methods Two hundred and thirty-eight stool specimens were collected from infant younger than 1 year old,that were hospitalized or outpatient from August to November 2015.Immunochromatography targeted GDH and toxin A&B of C.difficile was used for C.difficile screening,and those positive specimens were inoculated in CDIF and anaerobic culture.C.difficile isolates were genotyped by using slpA sequence typing (slpA ST),and tcdA,tcdB,cdtA and cdtB of C.difficile isolates were detected by PCR.Results Fifty C.difficile strains were isolated from 238 stool samples,and the isolated rates of C.difficile from <3 months,3 months to <6 months,and 6 months to 1 years old groups were 9.3%,17.6% and 27.3%(χ2=6.940,P=0.031<0.05),respectively.52.0%(26/50) of the C.difficile isolates were toxigenic,and 69.2% (18/26) toxigenic isolates harbored tcdA+tcdB+cdtA-cdtB-.Fifty C.difficile isolates were genotyped as 11 slpA STs,slpA ST fr-02 and kr-02 were the commonest genotypes in toxigenic C.difficile isolates;however,that was slpA ST xr-03 in non-toxigenic isolates.Conclusion High C.difficile carriage is found in infants younger than 1 year old,and more than half of C.difficile isolates are toxigenic.Most of toxigenic isolates harbored toxin A and B.The genotype of C.difficile isolates is different between toxigenic isolates and non-toxigenic isolates.

Objective:To analyze and evaluate the safety and efficacy of a Chinese domestically manufactured Heart Con-type implantable third-generation magnetic and hydrodynamic levitation left ventricular assist device(LVAD) for the treatment of end-stage heart failure(ESHF), by reporting the results of eleven-center clinical trial on 50 cases.Methods:This study was a multicenter clinical trial, designed by means of prospective, multicenter and single-group target value. 50 subjects with ESHF were competitively enrolled and treated with HeartCon as the LVAD in eleven centers. The primary efficacy measure was survival, defined as either the subjects experiencing the transition to heart transplantation(HT) or myocardial recovery assisted by the device within 90 days, or as successfully assisted by the LVAD for full 90 days after implantation. The target survival rate was 60%, other observations included implantation success rate, mortality, pump failure needing replacement or emergency heart transplantation.Results:All enrolled 50 patients received LVAD implantation successfully, 46 survived with the pump for 90 days, 1 patient transitioned to heart transplantation, and 3 patients experienced pump thrombosis, within which 2 patients underwent pump replacement and continued to live with the pump for 90 days, and the other one received emergency heart transplantation. There were no dropout subjects. The survival rate at full 90 days after HeartCon implantation was 100%. The survival rates with pump in the full set analysis and the protocol set analysis were 96.00% and 95.92% respectively, which were higher than the target value of 60%. The differences were both statistically significant( P<0.05). Conclusion:The results of the multicenter clinical trial with the largest sample size in China using domestically manufactured third-generation LVAD has demonstrated that, HeartCon is a safe and effective LVAD to treat ESHF patients.

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 application value of laparoscopic natural orifice specimen extraction surgery (NOSES) based on purse-string suture for sigmoid colon and upper rectal cancer.Methods:The retrospective and descriptive study was conducted. The clinicopathological data of 14 patients undergoing laparoscopic NOSES based on purse-string suture for sigmoid colon and upper rectal cancer in The First Affiliated Hospital of Sun Yat-sen University from October 2022 to June 2023 were collected. There were 8 males and 6 females, aged (56±10)years. Observation indicators: (1) surgical conditions; (2) postoperative conditions; (3) follow-up. Measurement data with normal distribution were represented as Mean± SD. Count data were described as absolute numbers. Results:(1) Surgical conditions. All patients underwent laparoscopic NOSES based on purse-string suture for sigmoid colon and upper rectal cancer successfully, without conversion to open surgery. The operation time of 14 patients was (162±32)minutes, and the volume of intraoperative blood loss was (22±12)mL. (2) Postoperative conditions. Time to postoperative first out-of-bed activity, time to postoperative first flatus, time to postoperative first drinking, time to postoperative initial liquid food intake, duration of postoperative hospital stay of 14 patients were (1.6±0.7)days, (2.1±0.6)days, (2.4±0.6)days, (3.8±1.0)days, (6.0±0.9)days, respectively. None of patient had perioperative complications such as postoperative anastomotic leakage and bleeding. (3) Follow-up. All 14 patients were followed up for (9.7±1.9)months. There was no postoperative recurrence, metastasis or death in 14 pati-ents.Conclusion:The laparoscopic NOSES based on purse-string suture can be used for sigmoid colon and upper rectal cancer, which is safe and feasible.

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.