Objective To screen the distribution frequency of Mur blood group among voluntary blood donors in Hezhou,Guangxi,and further analyze the molecular basis of of Mur antigen positive samples.Methods The Mur pheno-type of voluntary blood donors in Hezhou was serologically screened using microplate method,and the distribution frequency of Mur antigens in different ethnic groups was analyzed.Genetic typing was performed on these positive samples with PCR-SSP method to verify the accuracy of the serological method,and the genetic background was sequenced and analyzed.Re-sults Among 3 298 samples from voluntary blood donors in Hezhou,432(13.10%,432/3 298)were screened positive for Mur antigen,and PCR-SSP genotyping validation showed that all 432 samples were electrophoretic positive.Among them,the proportion of Han blood donors with positive Mur antigen was12.79%(331/2 587),Yao ethnic group was13.25%(64/483),Zhuang ethnic group was 16.51%(36/218),and no statistically significant difference was found in the three groups(P＞0.05).Further sequencing results showed that 428 samples were GYP(B-A-B)Mur,also known as GYP.Mur type(12.98%,428/3 298),the other 4 samples were GYP(B-A-B)Bun,also known as GYP.Bun type(0.12%,4/3 298).Conclusion The Mur blood type frequency is high in the voluntary blood donors in Hezhou,Guangxi,and is predominant characterized by GYP.Mur genotype.Due to ethnic integration,no significant difference was noticed in the frequency of Mur blood type distribution between Han,Zhuang and Yao population.Therefore,conducting extensive Mur blood group antigen and antibody testing in Hezhou is of great significance for ensuring clinical blood transfusion safety.

ObjectiveTo observe the effects of Cuscutae Semen on the learning and memory ability， N6-methyladenosine （m⁶A）-related modification enzymes and total m⁶A level in hippocampus of the offspring of fear-damaged pregnant rats. MethodForty-five pregnant rats were randomly divided into blank group， model group and Cuscutae Semen group. From the 1^st day to the 19^th day of pregnancy， rats in the model group and the Cuscutae Semen group were induced by observing electric shock of other rats. The Cuscutae Semen group was treated with 5 g·kg^-1·d^-1 Cuscutae Semen decoction （ig）， while the other two groups were treated with the same amount of purified water. The offspring were assigned following the grouping method of their maternal generation. The behavioral changes of the offspring were tested by Morris water maze on 21^st day after birth， and the development of hippocampal neurons was observed by transmission electron microscopy. The mRNA and protein expression levels of methyltransferase-like 3 （METTL3）， METTL14， Wilms tumor 1 associated protein （WTAP）， fat mass and obesity-associated protein （FTO） and Alk B homologue 5 （ALKBH5） were detected by Real-time polymerase chain reaction （Real-time PCR）， Western blot and immunohistochemistry （IHC）. The total content of m⁶A in hippocampus was determined by high performance liquid chromatography tandem mass spectrometry （LC-MS/MS）. ResultCompared with the conditions in the blank group， the average latency duration in the model group was prolonged， and the number of entries in the target quadrant， the target quadrant duration and the number of crossing the platform were decreased （P<0.01）. Additionally， the model group had seriously damaged structure of hippocampal CA1 and CA3 neurons， swollen mitochondria， expanded endoplasmic reticulum， and small number of synapses with some having blurred structure， and the expression levels of METTL3， METTL14， FTO， ALKBH5 as well as the total m⁶A level were lower than those in the blank group （P<0.05，P<0.01）. Compared with the model group， the Cuscutae Semen group had shortened average latency duration， increased number of entries in the target quadrant， target quadrant duration and number of crossing the platform （P<0.01）， alleviated damage of hippocampal CA1 and CA3 neurons， fine structure of mitochondrial and endoplasmic reticulum， and clear， intact and dense synapses. And the expression levels of METTL3， METTL14， FTO as well as the total level of m⁶A were up regulated， while the expression level of ALKBH5 was down regulated in the Cuscutae Semen group （P<0.05，P<0.01）. ConclusionCuscutae Semen improved the learning and memory ability of the offspring of the rats affected by fear damaging kidney during pregnancy， protected hippocampal neurons， and up-regulated the expression levels of METTL3， METTL14， FTO and the total m⁶A level in hippocampus.

Claudin18.2(CLDN18.2)is a tight junction protein that is overexpressed in a variety of solid tumors such as gastrointestinal cancer and oesophageal cancer.It has been identified as a promising target and a potential biomarker to diagnose tumor,evaluate efficacy,and determine patient prognosis.TST001 is a recombinant humanized CLDN18.2 antibody that selectively binds to the extracellular loop of human Claudin18.2.In this study,we constructed a solid target radionuclide zirconium-89(89Zr)labled-TST001 to detect the expression of in the human stomach cancer BGC823CLDN18.2 cell lines.The[89Zr]Zr-des-ferrioxamine(DFO)-TST001 showed high radiochemical purity(RCP,＞99％)and specific activity(24.15±1.34 GBq/μmol),and was stable in 5％human serum albumin,and phosphate buffer saline(＞85％RCP at 96 h).The EC50 values of TST001 and DFO-TST001 were as high as 0.413±0.055 and 0.361±0.058 nM(P＞0.05),respectively.The radiotracer had a significantly higher average standard uptake values in CLDN18.2-positive tumors than in CLDN18.2-negative tumors(1.11±0.02 vs.0.49±0.03,P=0.0016)2 days post injection(p.i.).BGC823CLDN18.2 mice models showed high tumor/muscle ratios 96 h p.i.with[89Zr]Zr-DFO-TST001 was much higher than those of the other imaging groups.Immunohistochemistry results showed that BGC823CLDN18.2 tumors were highly positive(+++)for CLDN18.2,while those in the BGC823 group did not express CLDN18.2(-).The results of ex vivo biodistribution studies showed that there was a higher distribution in the BGC823CLDN18.2 tumor bearing mice(2.05±0.16％ID/g)than BGC823 mice(0.69±0.02％ID/g)and blocking group(0.72±0.02％ID/g).A dosimetry estimation study showed that the effective dose of[89Zr]Zr-DFO-TST001 was 0.0705 mSv/MBq,which is within the range of acceptable doses for nuclear medicine research.Taken together,these re-sults suggest that Good Manufacturing Practices produced by this immuno-positron emission tomog-raphy probe can detect CLDN18.2-overexpressing tumors.

AIM: To investigate the effect of tetramethylpyrazine (TMP) on doxorubicin (Dox) induced cardiotoxicity and the role of 14-3-3γ/Bcl-2 protein expression. METHODS: Primary cultured cardiomyocytes were randomly divided into Control group, Dox group, Dox+TMP group and Dox+TMP+pAD/14-3-3γ-shRNA group. After 48 hours, the cell viability was detected by MST, the activity of LDH in culture medium, the activities of Caspase-3, SOD, GSH-Px and the content of MDA were detected; the expression of 14-3-3γ and mitochondrial Bcl-2 was detected by Western blot; ROS generation, mitochondrial membrane potential and mPTP opening were detected by flow cytometry; apoptosis was detected by TUNEL method. RESULTS: After Dox exposed for 48 hours, the viability of cardiomyocytes decreased significantly, the activity of LDH in culture medium increased, the activities of SOD and GSH-Px decreased, the content of MDA increased, ROS generation increased; the mitochondrial membrane potential decreased, mPTP continued to open, caspase-3 activity and apoptosis increased. TMP pretreatment significantly upregulated the expression of 14-3-3γ and mitochondrial Bcl-2, and reversed the above changes simultaneously; pAD/14-3-3γ-shRNA not only downregulated the expression of 14-3-3γ, but also decreased the expression of Bcl-2 in mitochondria. CONCLUSION: TMP pretreatment upregulates the expression of 14-3-3γ and mitochondrial Bcl-2, inhibits oxidative stress, maintains mitochondrial function and reduces Dox induced apoptosis.

BACKGROUND:Internal fixation is commonly used in an early stage of intertrochanteric fractures of the aged worldwide, and can apparently reduce complications and fatality rate. The commonly used internal fixators contain proximal femoral nail anti-rotation and dynamic hip screw, whose comparison is current research hotspot.
OBJECTIVE: To evaluate the hip function and stability after internal fixation with proximal femoral nail anti-rotation and dynamic hip screw in repair of intertrochanteric fractures of the aged.
METHODS:A total of 64 aged patients with intertrochanteric fractures were enroled in this study, and assigned to dynamic hip screw group (n=30) and proximal femoral nail anti-rotation group (n=34). The fracture reduction and healing were evaluated using anterioposterior and lateral X-ray films. Operation time, intraoperative blood loss, complication rate, healing time and postoperative hip function were compared and analyzed between the two groups, and then assessed in accordance with Sander’s scoring system.
RESULTS AND CONCLUSION:Compared with dynamic hip screw group, operation time, intraoperative blood loss, healing time and complication rate after treating unstable intertrochanteric fracture (Tronzo-Evans III, IV and V types) showed significant advantages in the proximal femoral nail anti-rotation group (P < 0.05). Hip function restored better, and the complication rate of stable intertrochanteric fracture (Tronzo-Evans I and II types) was lower after treatment in both groups (P > 0.05). These data show that the effects of proximal femoral nail anti-rotation were better than that of dynamic hip screw for treatment of intertrochanteric fractures, and hip function recovered better. Moreover, proximal femoral nail anti-rotation had biomechanical stability, especialy for unstable fracture.

Objective Breast intraductal papillary lesion ( BIPL) has a low incidence but a high rate of malignancy .This study discusses the diagnostic value of conventional ultrasonography ( US) combined with ultrasonic elastography ( UE) for breast papil-lary lesion. Methods We analyzed the preoperative ultrasound data of 48 patients with 63 BIPLs, and classified them according to the characteristics of two-dimensional ultrasound (2DUS) images.Then we compared their color Doppler ultrasound characteristics and UE features with the pathological results . Results Based on the 2DUS findings, the BIPLs were divided into 4 types.The sensitivi-ty, specificity, and accuracy of conventional US +UE in the diagnosis of BIPL were 93.2%, 88.9%, and 90.5%, respectively, markedly higher than those of conventional US (75.6%, 66.7%, and 73.0%), with statistically significant differences in the areas under the curve between the two methods (0.918 vs 0.838, P<0.05). Conclusion Conventional US combined with UE can im-prove the diagnosis of breast intraductal papillary lesion .

Objective To analyze and determine the efficient T- and B-combined (T/B) antigenic epitopes in Helicobacter pylori adhesin A. Methods Recombinant HpaA (rHpaA) was expressed for immunizing rabbit to generate antiserum. T- and B-cell epitopes in HpaA molecule were predicted by using bioinformatic technique. The segments to encode T/B combined epitope peptides were amplified by PCR and the phage display systems of T/B combined epitopes were subsequently constructed. PEG/NaCl precipitation method was applied to extract the recombinant phage PⅢ (rPⅢ) that displayed T/B combined epitopes. By using either commercial IgG against whole-cell of Helicobacter pylori or rHpaA antiserum as the primary antibody, the T/B combined epitopes displayed in rP Ⅲ s were screened and identified by Western blot and ELISA. MTT was applied to determine the proliferation of rHpaA-immunized mouse splenocytes after stimulation of the different recombinant rPⅢ proteins. Results In the HpaA molecule there were five T/B combined epitopes: HpaA10, HpaA37, HpaA79, HpaA116 and HpaA143. All the T/B combined epitopes were successfully displayed on the surface of PⅢ protein of phage M13. The results of Western blot, ELISA and MTT confirmed that HpaA116 was the predominant antigenic epitope, both HpaA37 and HpaA79 were the efficient antigenic epitopes. However, HpaA10 and HpaA143 were identified as ineffective antigenic epitopes. Conclusion The phage display systems of T/B combined epitope peptides of H. pylori adhesin A have been successfully generated in this study. HpaA37 and HpaA79, especially HpaA116 are the efficient T/B combined antigenic epitopes in HpaA of H. pylori.

Objective For better recognition of the hereditary nonpolyposis colorectal cancer (HNPCC) with reference to its clinical characteristics, diagnosis and treatment [WT5”HZ]Methods [WT5”BZ] A survey of 7 families of HNPCC patients was made [WT5”HZ] Results [WT5”BZ]Among the 7 families, 21 persons had colorectal cancer with age ranging from 26 to 79 years (average 46 years) The total of the colorectal cancer foci in 21 cases were 31,among which 16 were located proximal to,10 distal to the splenic flexure and 5 without definite record 61 5%(16/26) of the colorectal cancer foci were located proximal to the splenic flexure Six patients had multifocal colorectal cancer, which accounted for 28 6%(6/21) of all cases [WT5”HZ] Conclusions [WT5”BZ] HNPCC patients and their family members should be followed up regularly for early diagnosis and treatment of the disease and for better prognosis

Objective To investigate if the prognosis of young patients with colorectal cancer (CRC) is influenced by their familial predisposition. Methods The patients with CRC under 40 years old, underwent operation, had pathological diagnosis , family survey, followed up etc were selected into this research. According to familial predisposition they were divided into two groups: group A (with familial predisposition) and group B (without familial predisposition). Results Among the 44 patients twelve were in group A, thirty-two in group B. The 5-year survival rate in group A and B was 83.30%(10/12) and 59.38%(19/32) respectively,the difference was significant between the two groups(? 2=5.64,P

AIM: To determine the effect of heat treatment on rat primary cultured neurons, and give fundamental research for candidate molecule to protect the neurons from heat injury. METHODS: Neurons from rat striatum were primary cultivated in D-MEM with 15% horse serum, and when got mature, cells were identified by immuno-cytochemical staining with neurofilament protein (NF), tyrosine hydroxylase (TH) and neuron specific enolase (NSE) antibodies. Cells in heat treatment groups were put in an 43 ℃ CO 2 incubator for 1 h, and the control groups at 37 ℃ as normal. Striatum neurons were stained with trypan blue and dual fluorscence dye (PI/H33258) immediately followed heat treatment, and necrosis rate of neurons was estimated. At the same time, activated caspase-3 immuno-cytochemical and TdT TUNEL methods were applied to determine apoptosis rate, and cell volume was also identified with micro-photography. RESULTS: During day 7 to day 9, the cultured striatum neurons got mature, and many neuronal fibers starched out and formed neuron network, NF, TH, and NSE staining positive. Treatment at 43 ℃ for 1 h, cell number decreased greatly, while NF+ percentage kept unchanged, and the heat treatment survived neurons were processing cell necrosis and apoptosis, but necrosis percentage was much greater than that of apoptosis. While cell volume kept unchanged after heat treatment. CONCLUSION: Heat treatment greatly affects the growth and survival of the cultured striatum neurons, and the injury effect is most due to cell necrosis process.