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.

OBJECTIVE To provide a reference for continuing education and training of clinical pharmacists. METHODS The revision of the syllabus and the improvement of training methods of practical skills training class for clinical pharmacists in the neurology department held by Xuanwu Hospital of Capital Medical University from 2007 to 2022 was sorted to summarize its advantages and characteristics. RESULTS Training programs were developed to benefit clinical pharmacists at different levels， and the training contents were adjusted according to the training programs and the needs of trainees. Teachers with teaching experience were selected to participate in the teaching. Theory teaching was combined with practice teaching in the teaching process， and case teaching and question-based teaching methods were adopted to benefit both senior clinical pharmacists and new clinical pharmacists. In addition， the influence of the training class was expanded through online teaching， so that doctors and pharmacists could communicate and learn together on the platform of the training class. For example， when designing the training program， we replaced one common neurological disease every two years， and carried rollover study on its new progress and new ideas； clinical pharmacist skill course was reduced， drug history writing， information retrieval and test index interpretation were compressed into clinical pharmacy skill course. CONCLUSIONS The continuing education platform is established for clinical pharmacists； new knowledge and concepts that clinical pharmacists of this specialty need to be familiar with are compiled into the teaching syllabus， and the experts who are familiar with the training of clinical pharmacists are selected to explain to the students so that the students could follow the platform to constantly update their knowledge and improve the ability of clinical pharmacists to participate in the clinic work.

OBJECTIVE： To explore current hot spots in drug research and application，and provide reference of the draft use standard for pharmacy journal editor. METHODS： Taking China Pharmacy （short for our magazine） as example，all literatures published by our magazine and included by CNKI were searched during inception （Jan. 1990） and Mar. 19th， 2019； top 150 literatures [top 100 non-review literatures （NRA） and top 50 review literatures （RA）] were collected according to comprehensive score of downloaded number and citied number as indicators， and then analyzed in respects of publication year，column of NRA，first author’s work unit， subject words； one literatures which the cited number was largest was analyzed separately. RESULTS： Totally 150 literatures were published between 1993 and 2016， while the literature amount reached the maximum value in 2010 （18 articles）. Among NRA， the most were in the columns of “pharmacy administration” （44 articles， 44.00％）， “TCM and ethnic medicine” （18 articles， 18.00％）， “pharmacists and pharmaceutical care” （14 articles， 14.00％）. Among 150 first authors，73 （48.67％） were from college， 62 （41.33％） were from hospital. Subject words of NRA were “status and development” （20 times）， “active ingredient” （14 times）， “international experience” （9 times）， while those of RA were “pharmacological action” （25 times）， “Chinese herbal medicine” （15 times）， “active ingredient” （12 times）. The Principles and Utilization of Sequential Analysis of DDD System of Drug in Hospitals was cited for 1 426 times， which was much higher than the second one （261 times）. CONCLUSIONS： This study can show the good literatures origin and study direction，provide reference to the hot spots for readers. At the same time， high-frequency cited literatures also suggests that researchers can conduct in-depth analysis of controversial and unclear contents in academic circles.

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 improve the training quality of neurology clinical pharmacist. Methods:The experience during the training of neurology clinical pharmacists in recent ten years was summarized. Results:The students were taught in groups according to their majors and promoted in stages according to the teaching plans. Besides,various teaching methods were carried out timely. After tutored by the methods mentioned above,the students could quickly raise their working ability to play good roles of clinical pharmacist and work independently with more confidence. Conclusion:The training quality of neurology clinical pharmacists has been effectively improved by teaching in groups,promoting in stages and providing various teaching methods timely.

Objective:To explore the methods and the role of clinical pharmacists in the medical advice audit. Methods: The questionnaire was designed to analyze the results of the medical advice investigated by the clinical pharmacists from November 2011 to October 2012. Results:Totally 3 232 patients were in the use of antimicrobial drugs in a investigation of 14 675 patients in 178 724 medical advices, accounting for 22. 0% of the total number of the investigation;the use of antimicrobial agents in patients with medical advices of antimicrobial were 5 387, accounting for 3. 0% of the total number of the investigation;597 errors were detected and inter-vened, accounting for 0. 33% of the total number of the investigation;the main errors were dose errors, usage errors and repeated med-ication errors. Indication error and interaction error were more in the critically ill patients monitored by the clinical pharmacists than that in the general ward patients. Conclusion:Clinical pharmacists and ward pharmacists to participate medical advice review, partic-ularly to concern about the patients with medical advices of antimicrobial drugs, and to intervene reasonably in the dose errors, usage errors and repeated medication errors can improve the overall quality of the hospital pharmacist team, make sustainable development of rational use of drugs and reduce the risk of medication.

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 investigate the effect of intrathecal injection of Tanshinone Ⅱ A on bone cancer pain behavior and spinal IL-1 β,IL-6,TNF-α expression.Methods According to the random number table method,84 C3H/HeNCrlVr male mice were divided into:(1) Tanshinone ⅡA 10 μg group:the tumor mice were treated by intrathecal administration (once daily on the days 14-20 after inoculation of tumor cells) with Tanshinone ⅡA 10 μg; (2)Tanshinone ⅡA 20 μg group:the tumor mice were treated with Tanshinone ⅡA 20 μg; (3)Tanshinone ⅡA 40 μg group:the tumor mice were treated with Tanshinone ⅡA 40 μg; (4) normal control group:the mice were given food and water ad libitum; (5) DMSO+Sham group:the sham mice were treated with the same volume of 5％DMSO; (6) Tanshinone ⅡA+Sham group:the sham mice were treated with Tanshinone ⅡA 40 μg; (7)DMSO+Tumor group:the tumor mice were treated with the same volume of 5％DMSO.The mice pain behaviors were assessed with the paw withdrawal thermal latency (PWTL) at the corresponding time points,then the mice were killed and the samples of spinal cord were detected by real-time PCR.Results The basic values of PWTL had no significant differences among all groups (P＞0.05).At day 14 after operation,no significant difference (P＞0.05) was found in the PWTL value between normal control group and the sham operation group.But in tumor group,the PWTL value was significantly lower than that of normal control group (P＜0.05).At day 21 after operation,the PWTL and the level of spinal IL-1 β,IL-6,TNF-α expression had no significant differences (P＞0.05) among normal control group,Tanshinone ⅡA+Sham group and DMSO+Sham group.The PWTL ((6.19± 1.26)s) in DMSO+ Tumor group was significantly lower than that of normal control group((16.01± 1.59)s) (P＜0.05),but the level of IL-1β,IL-6,TNF-α expression was higher than that of normal control group.Compared with the normal group,the PWTL ((9.83±1.26)s;(10.29±2.95) s) of Tanshinone Ⅱ A 20 μg and 40 μg group was higher,and the level of spinal IL-1β,IL-6,TNF-α expression was lower (P＜0.05).The PWTL and the levels of spinal IL-1β,IL-6,TNF-αexpression had no significant differences between Tanshinone ⅡA 10 μg group ((6.67 ± 0.96) s) and DMSO + Tumor group(P＞0.05).Conclusion Intrathecal injection of Tanshinone Ⅱ A plays a role in anti-cancer pain,and inhibition of spinal inflammatory cytokine release may be one of its mechanisms.

AIM: To construct full length hCD59 eukaryotic and extracellular domain of hCD59 (hsCD59) prokaryotic expression vectors and prepare polyclonal antibody of hCD59. METHODS: cDNA fragments encoding hCD59 and hsCD59 were amplified from human PBMCs by RT-PCR and cloned into the eukaryotic vector pVAX-1 and prokaryotic vector pGEX-KG, respectively. The recombinant fusion protein GST-hsCD59 was expressed in E. coil BL21 induced by IPTG. Then the fusion protein was purified and identified. Polyclonal antibody against hCD59 was prepared by immunizing rabbit with pVAX-1-hCD59 and boosting with GSThsCD59 fusion protein, and the titer was identified. RESULTS: The recombinant eukaryotic vector pVAX-1-hCD59 and prokaryotic vector pGEX-KG-hsCD59 were successfully constructed. The GST-hsCD59 fusion protein was over-expressed in E. coli BL21 and the relative molecular mass (M~r) of the expression product was identical with predicted size. The titer of the anti-hCD59 serum was 1:3 200. CONCLUSION: We got the recombinant eukaryotic vector pVAX-1-hCD59, prokaryotic vector pGEX-KG-hsCD59 and rabbit anti-hCD59 polyclonal antibody successfully. These work would be helpful for the further study of the biological function of human CD59.