Objective To explore a venipuncture method that can reduce the pain, improve the suc-cess rate of puncture as well as reduce vascular injury. Methods Self-controlled method was applied in 200 patients, needle tips with 20～25°,31～40°,41～50°, 51～60°, > 60° angles on the top of the blood ves-sels,the inclined plane of the needle parallel to the longitudinal axis of the blood vessel (that was,the in-clined plane to the left or right) or traditional method with upward of needle tips were adopted, the observa-tion group and the control group were divided according to different methods, pain and success rate of puncture were compared. Results There were differences between the two groups,results of the observa-tion group were better than those of the control group (except when > 60°, success rate showed no differ-ence). Conclusions Venipuncture with needle tip parallel to the longitudinal axis of the blood vessels can significantly reduce pain in patients, improve the success rate of puncture and reduce the vascular in-jury, which were superior to the traditional venipuncture method. Whatever method is chosen, 51～60° is the best angie into the vessel.

Objective To investigate the influence and the clinical significance of interleukin-2 (IL-2) on T lymphocyte subgroup after chemotherapy in acute myeloid leukemia patients. Methods Fiftyfour acute myeloid leukemia patients were divided into treatment group (chemotherapy combined with IL-2,28 cases) and control group (pure chemotherapy,26 cases) by radom digits table. In treatment group, IL-2400 000 U was dripped after chemotherapy for 14 days. Then the T lymphocyte subgroup change before and after treatment was examined. Results After 14 days' treatment, the levels of CD3 (0.6026±0.2275 ),CD4(0.4972±0.1224),CD56(0.3016±0.1053 ) in treatment group were significantly higher than those before treatment(0.3926±0.2010,0.2264±0.1190,0.1729±0.1226) and in control group (0.4352±0.1930,0.2738±0.1362,0.1937±0.1268)(P< 0.05). Conclusion IL-2 treatment can obviously raise the levels of T lymphocyte subgroup and can raise the immunologic function.

Objective To evaluation the clinical implemented and primary experience of TME combined the double-stapling technique in low rectal carcinoma anus-preserving resection. Methods 22 cases of low rectal carcinoma (the distance between the inferior verge of rectal turnout and the anal verge were 4～7cm) were operated by TME combined the double-stapling technique. Results All of the 22 cases of TME combined double-stapling procedure were successful without operative mortality. Correlative complications including anastomotic leakage (3 cases), anastomotic stenosis (1 case). Temporary completely defunctioning loop colostomy 4 cases. Conclusions TME combined the double-stapling technique in low rectal carcinoma anus-preserving resection is feasibility and the tumour pathology and clinical stage should be considered. Mastering expert operation skill and adherence to sound surgical priciples is the key to prevent and to decrease postoperative complication.

Objective To investigate the relationship between Helicobacter prlori (Hp) infection and type 2 diabetes related complications.Methods Eighty-five patients with type 2 diabetes had been tested and divided into Hp negative group (42 cases) and Hp positive group (43 cases),the triglyceride (TG),low density lipoprotein cholesterol (LDL-C),fasting plasma glucose (FPG),glycosylated hemoglobin (HbA1c),tumor necrosis factor-α (TNF-α) and complications occurred were compared between two groups.Results The TG,LDL-C,FPG,HbA1c and TNF-α in Hp positive group was significantly higher than that in Hp negative group [(4.83 ± 0.95) mmol/L vs.(3.03 ± 0.75) mmol/L,(4.99 ± 0.96) mmol/L vs.(3.91 ±0.87)mmol/L,(10.85 ±2.97) mmol/L vs.(8.11 ±2.69) mmol/L,(9.35 ± 1.96)％ vs.(7.08 ± 1.53)％,(7.33 ± 0.78) ng/L vs.(3.65 ± 0.98) ng/L] (P＜ 0.01).The cardiovascular disease,cerebrovascular disease,kidney disease,retinal changes and podiatric complication rate in Hp positive group was 51.16％ (22/43),37.21％(16/43),27.91％(12/43),34.88％(15/43),23.26％(10/43),significantly higher than that in Hp negative group [28.57％ (12/42),16.67％ (7/42),9.52％ (4/42),14.29％ (6/42),7.14％ (3/42)],there was significant difference between two groups (P ＜ 0.05).Conclusion Type 2 diabetes Hp infection can make the higher TG,LDL-C,FPG,HbA1c,TNF-α and can increase the risk of diabetes complications.

Objective To study the protective effect of Res o n myocardial ischemic/reperfused induced injury. Methods My ocardial ischemic/reperfusion model was used to study the protective effect of d ifferent dosage resvaratrol on myocardial ischemic/reperfusion injury in rats. Results Res shrinked the size of myocardial infarction indu ced by the ischemic reperfused method, inhibited the release of creatine kinase( CK) and lactate dehydrogenase(LDH) from the injured myocardium and reduced elega ted ST-T of electrocardiogram(ECG) caused by myocardial ischemic-reperfused in jury in dose-dependent way. Res also improved the morphological changes of inju red myocardium. Conclusion Res has protective action on myo cardial ischemic reperfused injury in rats.

Objective To investigate the therapeutic effect of VSD combined with external fixator in the treatment of open tibial and fibular fractures in Tibet plateau. Methods From August 2014 to August 2015, totally 16 cases of open tibial and fibular fractures were treated in our department, including 12 males and 4 females with an average 39. 4 years-old. There were 4 cases of proximal and mid tibial fractures while 12 cases were distal fractures. Debridement, vacuum sealing drainage combined with external fixator for stabilization of fractures was performed in all patients after general condition was improved. A vacuum sealing drainage change or second stage wound closure or soft tissue coverage was undergone after 7 days. Bone union time by X-ray and complications were recorded. Results All wounds were healed by second stage. Twelve patients months were followed up ( by calling back to the hospital ) for an average 18 months ( 12 to 24 months ) , 4 cases were lost to follow-up. X-ray demonstrated that bone union was obtained at a mean 5. 5th month (3 to 7 months). There were 9 cases (75%) obtained a first phase of fracture healing, while 3 cases ( 25%) of delayed healing. Pin-track infection was occurred in two patients and was cured by conservative treatment. No complications of deep infection, graft skin or flap necrosis, malunion, nonunion or osteomyelitis was seen during follow-up. Conclusions Vacuum sealing drainage combined with external fixator for treatment of open tibial and fibular fracture in Tibet can rapidly and effectively stabilize the fracture, and simultaneously safely and effectively seal the wound, reduce the wound healing time, promote fracture healing and lower the complication rates.

OBJECTIVE To study the effect of partial ossicular replacement prothesis (PORP) on the middle ear transfer function.METHODS On a incus removed finite element model,a titanium columnar structure which link the malleus and the stapes was used to simulate the function of PORP to transmit acoustic pressure.The effect of placement of PORP on different positions of the manubrium,as well as the insertion of the cartilage between the artificial bone and the fascia on the middle ear transfer function were calculated.RESULTS When the PORP is placed on the neck of manubrium,mid-manubrium and headof malleus removed,the vibration amplitude of the umbo ofthe tympanic membrane is reduced in 200-8000 Hz,and thereduction range in 200-3200 Hz is more than that of 3200-8000 Hz.While the vibration amplitude of the stapes in the200-8000 Hz amplitude increased.The vibration amplitude is basically the same when placed on the neck of manubrium and mid-manubrium,while it slightly increased about 2-3 dB than the former two when the head of malleus removed.In 1000-8000 Hz.When putting the cartilage between the PORP and transplanted fascia,the vibration amplitude of the stapes footplate was slightly increased,but there was a slight difference with different cartilages.CONCLUSION The placement and material property of PORP play an important role in the middle ear transfer function.

An analytical method was developed for the simultaneous extraction and determination of three quaternary ammonium compouds ( QACs) in soil samples using ultrasonic extraction and gas chromatography-mass spectrometry( GC-MS) . The qualitative and quantitative analysis of the three analytes dodecyltrimethyl ammonium chloride ( DTAC) , cetyltrimethyl ammonium bromide ( CTAB) and didodecyldimethyl ammonium chloride ( DDAC) were conducted by application of EI mass spectra and selected ion monioring ( SIM ) . Characteristic ions of the QACs were m/z 58 ( DTAC and CTAB) and m/z 212 ( DDAC) . To achieve optimum extraction efficiency, several impact factors including types of extractants, pH of extraction, concentration of linear alkylbenzene sulfonates ( LAS) , extraction times and content of purification column were investigated. Methanol with pH 3. 5 and 40 μg/L LAS solution were selected as extractant. Soil sample was extracted by treated methanol each 10 mL for 20 min every time. Extract of the soil sample was purified by neutral alumina column with 4 cm in length and 1cm in diameter, and then was determineted by GC-MS. Good linear relationships of all the three QACs were obtained in the range of 0. 02-2. 0 mg/L. The limits of determination (LOD, S/N=3) was 1. 2-4. 5 μg/kg. The method was used to analyse real soil samples ( paddy soil, lateritic red soil, and ore tailings) collected from a mining district in south China. Results of determination exhibited the concerntrations of the three analytes in real soil samples ranged from 0 . 24 mg/kg to 0. 41 mg/kg, and their recoveries ranged from 76% to 113% with relative standard deviations ( RSD) of 1. 1%-12. 9% in three different spiked concentrations of 0. 2, 0. 5 and 1. 0 mg/kg.

Objective To perform laboratory detection and molecular traceability analysis on a case of imported kala-azar in Shenzhen to determine the infection strain. Methods Bone marrow puncture fluid and blood samples from a case of kala-azar in Shenzhen were collected for laboratory tests. The patient's bone marrow puncture fluid smears were stained with Giemsa and examined under a microscope. Blood samples were examined for antibodies using the rk39 visceral leishmania rapid diagnostic reagent. Whole blood DNA was extracted, and the ITS-1 sequence was amplified by PCR, sequenced and aligned, and a phylogenetic tree was constructed based on the ITS-1 sequence. Results Microscopic examination of the patient's bone marrow smears revealed a large number of Leishmania amastigotes without flagella, confirming the diagnosis of kala-azar. The patient's blood was tested positive with the rk39 rapid diagnostic reagent, and PCR amplification yielded an ITS-1 gene product sequence that matched the expected size. Sequence alignment with the NCBI database showed 100% sequence similarity with the ITS-1 gene sequence of Leishmania infantum, confirming the infecting strain as Leishmania infantum. Phylogenetic tree construction of the amplified ITS-1 sequence revealed clustering into a clade with Leishmania infantum , and close to KC347299, one of the reference sequence selected. Conclusions The case of kala-azar in Shenzhen was caused by Leishmania infantum. Kala-azar still occurs in China, so the diagnostic technology of medical personnel in non-epidemic areas should be strengthened so that they can actively use new diagnostic technologies to assist in diagnosis, thus improving their prevention and control ability of Leishmania parasites.

Abstract: Objective　To analyze and understand the mutations of drug resistance genes in imported Plasmodium falciparum in Shenzhen, aiming to assess the efficacy of antimalarial drugs and guide effective drug use. Methods A total of 85 samples from individuals with imported Plasmodium falciparum confirmed by fluorescence quantitative polymerase chain reaction (PCR) in Shenzhen from 2022 to 2023 were collected and genomic DNA was extracted. Nested PCR was used to amplify resistance genes, including Plasmodium falciparum Kelch 13 (PfK13), multidrug resistance gene 1 (Pfmdr1), chloroquine resistance transporter (Pfcrt), dihydrofolate reductase (Pfdhfr), and dihydropteroate synthase (Pfdhps) genes. Bidirectional sequencing was conducted, and mutations in these resistance genes were analyzed using MEGA11.06 software. Results　The study found one missense mutation (S549P) and four synonymous mutations in PfK13. For Pfmdr1, 62.69% of the samples showed Y184F mutation, and no N86Y mutation was detected. No mutations at positions 72 and 73 were detected in the Pfcrt gene, while mutations at M74I, N75E, and K76T accounted for 17.46%, 15.87%, and 15.87%, respectively. The wild-type of Pfcrt gene is dominant (82.54%, 52), followed by the triple mutant I74E75T76 (15.87%, 10). The most common mutation type for Pfdhfr is I51R59N108 (91.78%, 67), followed by the wild type (2.74%, 2). More than half (60.32%, 38) of the Pfdhps samples were wild-type, with single mutation K540E being the most common mutation type. S436A, G437A, K540E, A581G, A613S, I431V, G556K, and G579E site mutations were detected. Among the Pfdhfr-Pfdhps combination mutations, I51R59N108-E540 was the most frequent combination mutation (11.48%), with 59.02% of samples showing solitary Pfdhfr mutations. Conclusions In this study, PfK13 mutation rates were low, with no reported resistance mutations. The Y184F mutation emerged as the dominant Pfmdr1 mutation, with no detection of N86Y. For Pfcrt, the wild-type was dominant, followed by the I74E75T76 triple mutation variant. Triple mutant I51R59N108 of Pfdhfr was very common, and our study did not find Pfdhfr Pfdhps completely resistant and super resistant mutants, but there were other quintuple and septuple mutant types. In the future, it is crucial to continue to strengthen the monitoring of malaria parasite resistance genes and to further integrate in vivo efficacy monitoring to effectively guide clinical drug use.